laboratory diagnostic of invasive fungal infections · serious infection •isolation of candida sp...
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LABORATORY DIAGNOSTIC OF INVASIVE FUNGAL INFECTIONS
Mardiastuti H Wahid Department of Microbiology
Faculty of Medicine Universitas Indonesia
INTRODUCTION
Increased incidence of IFI
due to increased immuno-
compromised condition
malignancies
Increased resistance
to antimycotic
agents
The need for
accurate laboratory diagnostics
Better treatment and clinical outcome
LABORATORY DIAGNOSTIC OF IFI
bull (13)-β-D-Glucan assay accurate laboratory diagnostic tool in determining between definitive and probable invasive fungal infections
bull May be useful in the clinical practice if
ndash Implemented in a proper setting
ndash Interpreted after consideration of its limitation
Drosos E Karageorgopoulos et al Clinical Infectious Diseases 201152(6)750ndash770
bull The Glucatell serum BG detection assay is highly sensitive and specific as a diagnostic adjunct for IFI
Zekaver Odabasi et al
Clinical Infectious Diseases 2004 39199ndash205
LABORATORY DIAGNOSTIC OF IFI
INVASIVE CANDIDOSIS
SPECIES DISTRIBUTION
bull The incidence of Candidosis infection in hospitalized patients in the USA has increased (1980-1989) 8 responsible for all bloodstream infections
bull In the 1990s incidence of Candida bloodstream infections decreased due to a decline of C albicans infections however the incidence rate of C glabrata increased
bull 1992-1993 1998-2000 overall incidence of Candida bloodstream infections increased
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
bull Invasive candidosis
ndash endogenous
ndash transmission from person to person (direct patient-patient transmission healthcare workers medical devices contaminated intravenous infusions)
bull Cross infection in ICU
ndash C albicans via hand of healthcare workers
ndash C parapsilosis via medical devices
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
bull Invasive candidosis
ndash Neutropenic cancer patients
ndash Stem cellorgan transplants recipients
ndash ICU patients (adult surgical neonatal)
Widespread use of fluconazole emergence of less susceptible species
Note Intrinsic resistance (C glabrata and C krusei) to fluconazole factor in their emergence as pathogens
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
LABORATORY DIAGNOSTIC
bull Diagnosis of invasive candidosis is difficult due to a variation and non specific clinical presentation difficulties in intrepreting laboratory examination results
bull Dignosis based on clinical radiological microbiological and histopathological findings
bull Microbiology positive blood culture budding yeast cellspseudohyphae in specimens obtained from normal sterile or abnormal sites (biopsy aspiration)
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
LABORATORY DIAGNOSTIC
bull Investigation included
ndash Microscopy
ndash Culture
ndash Serological test
ndash D-arabinitol detection
ndash Molecular assays
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
MICROSCOPY
bull Typical budding yeast cells pseudohypahetrue hyphae of Candida species found in tissue biopsy normal sterile body fluids
bull C glabrata produces only yeast cells and
C albicans produces true hyphae in tissues
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
CULTURE
bull Isolation of Candida sp from sputum and faecal specimens can not be considered diagnostic of infection
bull Deep-seated infection Candida sp isolated from blood or normally sterile body fluids tissuesaspirates from other closed sites
bull Specimens should be examined as soon as possible after collection to avoid missinterpretation due to other organisms multiplication
bull Different susceptibility among Candida sp completed identification (speciation) before treatment
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
CULTURE
bull Blood cultures are positive in no more than 50 of neutropenic patients with dissemintaed candidosis or 80 of patients with endocarditis
bull Isolation of Candida sp from urine may indicate a serious infection
bull Isolation of Candida sp from LCS evidence for diagnosis of meningitis
bull Candida sp isolated from BAL in a patient with pulmonary infiltrates not sufficient for diagnosing candidosis
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
SEROLOGICAL TESTS
bull Limited usefulness
bull False positive (mucosal colonization superficial infection)
bull False negative (immunocompromised)
bull Antigen detection mannan insensitive
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
DETECTION OF D-ARABINITOL
bull D-arabinitol is produced by most of Candida sp (except C krusei and may be C glabrata)
bull It can be detected in serum or urine
bull Candidemia increased of serum D-arabiniitol-creatinine ratios has still to achieve widespread clinical use
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
MOLECULAR ASSAY
bull PCR detection of Candida DNA in blood serum LCS other specimens
bull Promising tools for rapid diagnosis of Candida sp from tissue or body-fluid
bull False positive
bull Standardized commercial assays were not available
bull Can not be recommended in diagnosis of invasive candidosis
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
The sources of yeast isolated
bull Sputum 49
bull Urine 13
bull Vaginal swab 9
bull Blood 8
bull Bronchial lavage 7
2010
bull Sputum 62
bull Vaginal swab 16
bull Blood 14
bull Urine 9
bull Bronchial lavage 5
2011
bull Sputum 102
bull Urine 18
bull Blood16 bull Bronchial lavage 12 bull Vaginal swab 11
2012
Candida spp isolated from blood
2010 2011 2012
C albicans 1 1 3
C parapsilosis 3 4 1
C pelliculosis 1 0 0
C lusitaniae 1 0 0
C tropicalis 0 2 10
C glabrata 0 1 0
C gulliermondii 0 1 1
Candida spp 1 0 0
7 9 15
Resistance pattern of Candida spp isolated from blood
2010 2011 2012
FCA ITR VRC FCA ITR VRC FCA ITR VRC
C albicans 17 115 115 115
C glabrata 19
C tropicalis 215 315 115
INVASIVE ASPERGILLOSIS
bull Incidence varies in ICU 58
bull In ICU risk factors including chronic obstructive pulmonary diseases and liver failure
bull Positive respiratory culture uncertain
bull Low sensitivity and specificity
bull Detection of galactomannan in BAL promising results
LABORATORY DIAGNOSTIC
bull Microscopy
bull Culture
bull Serological tests
bull Molecular diagnostics
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
MICROSCOPY
bull Histopathological examination of tissue biopsies immunohistochemical staining procedure
bull Non-pigmented septate hyphae dichotomous branching
bull Direct examination of sputum may help in diagnosing invasive aspergillosis BAL is better
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
A fumigatus
Richard C Barton Hindawi Publishing Corporation Scienti1048853 ca Volume 2013 Article ID 459405 29 pages httpdxdoiorg1011552013459405
CULTURE
bull Not sensitive interpreted with caution (contamination)
bull Recovered from blood LCS
bull Aspergillus isolated from BALsputum obtained from patients with a pulmonary infiltrates indicative of infection
bull A fumigatus can be isolated from sputum of patients suffer from allergic bronchopulmonary aspergillosis
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
SEROLOGICAL TESTS
bull Antobodies againts Aspergillus often helpful in the diagnosis of aspergillosis in non immunocompromised patients
bull Immunodiffusion indirect haemagglutination ELISA
bull The role is still uncertain
bull Positive ID test is not a proof of infection negative result can not exclude the diagnosis
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
SEROLOGICAL TESTS
bull Antigen detection (galactomannan) in blood and other body fluids rapid diagnosis of aspergilllosis in immunocompromised patients
bull Galactomannan fluctuate during the course of the infection useful if performed in a regular basis (twice per week)
bull Negative not exclude the diagnosis
bull ELISA for the detection of galactomannan has been approved for diagnostic use (the accuracy was moderate)
Christopher D Pfeiffer et al 2006
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
MOLECULAR DIAGNOSTICS
bull PCR for detection of Aspergillus in blood BAL serum etc
bull Appear promising for rapid detection of Aspergillus directly on tissue or body fluids but
ndash False positive
ndash No standardized commercial method
bull Routine use of PCR can not be recommended
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
Mark Reinwald et al
bull ldquoTreatment with mould-active antifungals prior to BAL sampling significantly decreases the performance of the Aspergillus PCR assay in haematological patients if BAL was performed after administration of more than one antifungal agentrdquo
bull ldquoThe effect of antifungal therapy on other surrogate parameters suggests reduced sensitivity rates with antifungal therapy for GM whereas no influence of antifungal therapy on β-D-glucan performance in a (single-centre retrospective) study was describedrdquo
J Antimicrob Chemother 2012 67 2260ndash2267
CONCLUSION
bull Various detection methods of agents responsible for invasive fungal infections have been developed
bull The lack of sensitivity and specificity of the test uncertainty in diagnosing IFI decrease successful treatment
bull Need further studies to improve and develop laboratory diagnostic tool for IFI
INTRODUCTION
Increased incidence of IFI
due to increased immuno-
compromised condition
malignancies
Increased resistance
to antimycotic
agents
The need for
accurate laboratory diagnostics
Better treatment and clinical outcome
LABORATORY DIAGNOSTIC OF IFI
bull (13)-β-D-Glucan assay accurate laboratory diagnostic tool in determining between definitive and probable invasive fungal infections
bull May be useful in the clinical practice if
ndash Implemented in a proper setting
ndash Interpreted after consideration of its limitation
Drosos E Karageorgopoulos et al Clinical Infectious Diseases 201152(6)750ndash770
bull The Glucatell serum BG detection assay is highly sensitive and specific as a diagnostic adjunct for IFI
Zekaver Odabasi et al
Clinical Infectious Diseases 2004 39199ndash205
LABORATORY DIAGNOSTIC OF IFI
INVASIVE CANDIDOSIS
SPECIES DISTRIBUTION
bull The incidence of Candidosis infection in hospitalized patients in the USA has increased (1980-1989) 8 responsible for all bloodstream infections
bull In the 1990s incidence of Candida bloodstream infections decreased due to a decline of C albicans infections however the incidence rate of C glabrata increased
bull 1992-1993 1998-2000 overall incidence of Candida bloodstream infections increased
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
bull Invasive candidosis
ndash endogenous
ndash transmission from person to person (direct patient-patient transmission healthcare workers medical devices contaminated intravenous infusions)
bull Cross infection in ICU
ndash C albicans via hand of healthcare workers
ndash C parapsilosis via medical devices
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
bull Invasive candidosis
ndash Neutropenic cancer patients
ndash Stem cellorgan transplants recipients
ndash ICU patients (adult surgical neonatal)
Widespread use of fluconazole emergence of less susceptible species
Note Intrinsic resistance (C glabrata and C krusei) to fluconazole factor in their emergence as pathogens
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
LABORATORY DIAGNOSTIC
bull Diagnosis of invasive candidosis is difficult due to a variation and non specific clinical presentation difficulties in intrepreting laboratory examination results
bull Dignosis based on clinical radiological microbiological and histopathological findings
bull Microbiology positive blood culture budding yeast cellspseudohyphae in specimens obtained from normal sterile or abnormal sites (biopsy aspiration)
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
LABORATORY DIAGNOSTIC
bull Investigation included
ndash Microscopy
ndash Culture
ndash Serological test
ndash D-arabinitol detection
ndash Molecular assays
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
MICROSCOPY
bull Typical budding yeast cells pseudohypahetrue hyphae of Candida species found in tissue biopsy normal sterile body fluids
bull C glabrata produces only yeast cells and
C albicans produces true hyphae in tissues
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
CULTURE
bull Isolation of Candida sp from sputum and faecal specimens can not be considered diagnostic of infection
bull Deep-seated infection Candida sp isolated from blood or normally sterile body fluids tissuesaspirates from other closed sites
bull Specimens should be examined as soon as possible after collection to avoid missinterpretation due to other organisms multiplication
bull Different susceptibility among Candida sp completed identification (speciation) before treatment
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
CULTURE
bull Blood cultures are positive in no more than 50 of neutropenic patients with dissemintaed candidosis or 80 of patients with endocarditis
bull Isolation of Candida sp from urine may indicate a serious infection
bull Isolation of Candida sp from LCS evidence for diagnosis of meningitis
bull Candida sp isolated from BAL in a patient with pulmonary infiltrates not sufficient for diagnosing candidosis
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
SEROLOGICAL TESTS
bull Limited usefulness
bull False positive (mucosal colonization superficial infection)
bull False negative (immunocompromised)
bull Antigen detection mannan insensitive
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
DETECTION OF D-ARABINITOL
bull D-arabinitol is produced by most of Candida sp (except C krusei and may be C glabrata)
bull It can be detected in serum or urine
bull Candidemia increased of serum D-arabiniitol-creatinine ratios has still to achieve widespread clinical use
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
MOLECULAR ASSAY
bull PCR detection of Candida DNA in blood serum LCS other specimens
bull Promising tools for rapid diagnosis of Candida sp from tissue or body-fluid
bull False positive
bull Standardized commercial assays were not available
bull Can not be recommended in diagnosis of invasive candidosis
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
The sources of yeast isolated
bull Sputum 49
bull Urine 13
bull Vaginal swab 9
bull Blood 8
bull Bronchial lavage 7
2010
bull Sputum 62
bull Vaginal swab 16
bull Blood 14
bull Urine 9
bull Bronchial lavage 5
2011
bull Sputum 102
bull Urine 18
bull Blood16 bull Bronchial lavage 12 bull Vaginal swab 11
2012
Candida spp isolated from blood
2010 2011 2012
C albicans 1 1 3
C parapsilosis 3 4 1
C pelliculosis 1 0 0
C lusitaniae 1 0 0
C tropicalis 0 2 10
C glabrata 0 1 0
C gulliermondii 0 1 1
Candida spp 1 0 0
7 9 15
Resistance pattern of Candida spp isolated from blood
2010 2011 2012
FCA ITR VRC FCA ITR VRC FCA ITR VRC
C albicans 17 115 115 115
C glabrata 19
C tropicalis 215 315 115
INVASIVE ASPERGILLOSIS
bull Incidence varies in ICU 58
bull In ICU risk factors including chronic obstructive pulmonary diseases and liver failure
bull Positive respiratory culture uncertain
bull Low sensitivity and specificity
bull Detection of galactomannan in BAL promising results
LABORATORY DIAGNOSTIC
bull Microscopy
bull Culture
bull Serological tests
bull Molecular diagnostics
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
MICROSCOPY
bull Histopathological examination of tissue biopsies immunohistochemical staining procedure
bull Non-pigmented septate hyphae dichotomous branching
bull Direct examination of sputum may help in diagnosing invasive aspergillosis BAL is better
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
A fumigatus
Richard C Barton Hindawi Publishing Corporation Scienti1048853 ca Volume 2013 Article ID 459405 29 pages httpdxdoiorg1011552013459405
CULTURE
bull Not sensitive interpreted with caution (contamination)
bull Recovered from blood LCS
bull Aspergillus isolated from BALsputum obtained from patients with a pulmonary infiltrates indicative of infection
bull A fumigatus can be isolated from sputum of patients suffer from allergic bronchopulmonary aspergillosis
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
SEROLOGICAL TESTS
bull Antobodies againts Aspergillus often helpful in the diagnosis of aspergillosis in non immunocompromised patients
bull Immunodiffusion indirect haemagglutination ELISA
bull The role is still uncertain
bull Positive ID test is not a proof of infection negative result can not exclude the diagnosis
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
SEROLOGICAL TESTS
bull Antigen detection (galactomannan) in blood and other body fluids rapid diagnosis of aspergilllosis in immunocompromised patients
bull Galactomannan fluctuate during the course of the infection useful if performed in a regular basis (twice per week)
bull Negative not exclude the diagnosis
bull ELISA for the detection of galactomannan has been approved for diagnostic use (the accuracy was moderate)
Christopher D Pfeiffer et al 2006
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
MOLECULAR DIAGNOSTICS
bull PCR for detection of Aspergillus in blood BAL serum etc
bull Appear promising for rapid detection of Aspergillus directly on tissue or body fluids but
ndash False positive
ndash No standardized commercial method
bull Routine use of PCR can not be recommended
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
Mark Reinwald et al
bull ldquoTreatment with mould-active antifungals prior to BAL sampling significantly decreases the performance of the Aspergillus PCR assay in haematological patients if BAL was performed after administration of more than one antifungal agentrdquo
bull ldquoThe effect of antifungal therapy on other surrogate parameters suggests reduced sensitivity rates with antifungal therapy for GM whereas no influence of antifungal therapy on β-D-glucan performance in a (single-centre retrospective) study was describedrdquo
J Antimicrob Chemother 2012 67 2260ndash2267
CONCLUSION
bull Various detection methods of agents responsible for invasive fungal infections have been developed
bull The lack of sensitivity and specificity of the test uncertainty in diagnosing IFI decrease successful treatment
bull Need further studies to improve and develop laboratory diagnostic tool for IFI
Increased incidence of IFI
due to increased immuno-
compromised condition
malignancies
Increased resistance
to antimycotic
agents
The need for
accurate laboratory diagnostics
Better treatment and clinical outcome
LABORATORY DIAGNOSTIC OF IFI
bull (13)-β-D-Glucan assay accurate laboratory diagnostic tool in determining between definitive and probable invasive fungal infections
bull May be useful in the clinical practice if
ndash Implemented in a proper setting
ndash Interpreted after consideration of its limitation
Drosos E Karageorgopoulos et al Clinical Infectious Diseases 201152(6)750ndash770
bull The Glucatell serum BG detection assay is highly sensitive and specific as a diagnostic adjunct for IFI
Zekaver Odabasi et al
Clinical Infectious Diseases 2004 39199ndash205
LABORATORY DIAGNOSTIC OF IFI
INVASIVE CANDIDOSIS
SPECIES DISTRIBUTION
bull The incidence of Candidosis infection in hospitalized patients in the USA has increased (1980-1989) 8 responsible for all bloodstream infections
bull In the 1990s incidence of Candida bloodstream infections decreased due to a decline of C albicans infections however the incidence rate of C glabrata increased
bull 1992-1993 1998-2000 overall incidence of Candida bloodstream infections increased
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
bull Invasive candidosis
ndash endogenous
ndash transmission from person to person (direct patient-patient transmission healthcare workers medical devices contaminated intravenous infusions)
bull Cross infection in ICU
ndash C albicans via hand of healthcare workers
ndash C parapsilosis via medical devices
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
bull Invasive candidosis
ndash Neutropenic cancer patients
ndash Stem cellorgan transplants recipients
ndash ICU patients (adult surgical neonatal)
Widespread use of fluconazole emergence of less susceptible species
Note Intrinsic resistance (C glabrata and C krusei) to fluconazole factor in their emergence as pathogens
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
LABORATORY DIAGNOSTIC
bull Diagnosis of invasive candidosis is difficult due to a variation and non specific clinical presentation difficulties in intrepreting laboratory examination results
bull Dignosis based on clinical radiological microbiological and histopathological findings
bull Microbiology positive blood culture budding yeast cellspseudohyphae in specimens obtained from normal sterile or abnormal sites (biopsy aspiration)
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
LABORATORY DIAGNOSTIC
bull Investigation included
ndash Microscopy
ndash Culture
ndash Serological test
ndash D-arabinitol detection
ndash Molecular assays
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
MICROSCOPY
bull Typical budding yeast cells pseudohypahetrue hyphae of Candida species found in tissue biopsy normal sterile body fluids
bull C glabrata produces only yeast cells and
C albicans produces true hyphae in tissues
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
CULTURE
bull Isolation of Candida sp from sputum and faecal specimens can not be considered diagnostic of infection
bull Deep-seated infection Candida sp isolated from blood or normally sterile body fluids tissuesaspirates from other closed sites
bull Specimens should be examined as soon as possible after collection to avoid missinterpretation due to other organisms multiplication
bull Different susceptibility among Candida sp completed identification (speciation) before treatment
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
CULTURE
bull Blood cultures are positive in no more than 50 of neutropenic patients with dissemintaed candidosis or 80 of patients with endocarditis
bull Isolation of Candida sp from urine may indicate a serious infection
bull Isolation of Candida sp from LCS evidence for diagnosis of meningitis
bull Candida sp isolated from BAL in a patient with pulmonary infiltrates not sufficient for diagnosing candidosis
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
SEROLOGICAL TESTS
bull Limited usefulness
bull False positive (mucosal colonization superficial infection)
bull False negative (immunocompromised)
bull Antigen detection mannan insensitive
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
DETECTION OF D-ARABINITOL
bull D-arabinitol is produced by most of Candida sp (except C krusei and may be C glabrata)
bull It can be detected in serum or urine
bull Candidemia increased of serum D-arabiniitol-creatinine ratios has still to achieve widespread clinical use
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
MOLECULAR ASSAY
bull PCR detection of Candida DNA in blood serum LCS other specimens
bull Promising tools for rapid diagnosis of Candida sp from tissue or body-fluid
bull False positive
bull Standardized commercial assays were not available
bull Can not be recommended in diagnosis of invasive candidosis
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
The sources of yeast isolated
bull Sputum 49
bull Urine 13
bull Vaginal swab 9
bull Blood 8
bull Bronchial lavage 7
2010
bull Sputum 62
bull Vaginal swab 16
bull Blood 14
bull Urine 9
bull Bronchial lavage 5
2011
bull Sputum 102
bull Urine 18
bull Blood16 bull Bronchial lavage 12 bull Vaginal swab 11
2012
Candida spp isolated from blood
2010 2011 2012
C albicans 1 1 3
C parapsilosis 3 4 1
C pelliculosis 1 0 0
C lusitaniae 1 0 0
C tropicalis 0 2 10
C glabrata 0 1 0
C gulliermondii 0 1 1
Candida spp 1 0 0
7 9 15
Resistance pattern of Candida spp isolated from blood
2010 2011 2012
FCA ITR VRC FCA ITR VRC FCA ITR VRC
C albicans 17 115 115 115
C glabrata 19
C tropicalis 215 315 115
INVASIVE ASPERGILLOSIS
bull Incidence varies in ICU 58
bull In ICU risk factors including chronic obstructive pulmonary diseases and liver failure
bull Positive respiratory culture uncertain
bull Low sensitivity and specificity
bull Detection of galactomannan in BAL promising results
LABORATORY DIAGNOSTIC
bull Microscopy
bull Culture
bull Serological tests
bull Molecular diagnostics
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
MICROSCOPY
bull Histopathological examination of tissue biopsies immunohistochemical staining procedure
bull Non-pigmented septate hyphae dichotomous branching
bull Direct examination of sputum may help in diagnosing invasive aspergillosis BAL is better
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
A fumigatus
Richard C Barton Hindawi Publishing Corporation Scienti1048853 ca Volume 2013 Article ID 459405 29 pages httpdxdoiorg1011552013459405
CULTURE
bull Not sensitive interpreted with caution (contamination)
bull Recovered from blood LCS
bull Aspergillus isolated from BALsputum obtained from patients with a pulmonary infiltrates indicative of infection
bull A fumigatus can be isolated from sputum of patients suffer from allergic bronchopulmonary aspergillosis
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
SEROLOGICAL TESTS
bull Antobodies againts Aspergillus often helpful in the diagnosis of aspergillosis in non immunocompromised patients
bull Immunodiffusion indirect haemagglutination ELISA
bull The role is still uncertain
bull Positive ID test is not a proof of infection negative result can not exclude the diagnosis
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
SEROLOGICAL TESTS
bull Antigen detection (galactomannan) in blood and other body fluids rapid diagnosis of aspergilllosis in immunocompromised patients
bull Galactomannan fluctuate during the course of the infection useful if performed in a regular basis (twice per week)
bull Negative not exclude the diagnosis
bull ELISA for the detection of galactomannan has been approved for diagnostic use (the accuracy was moderate)
Christopher D Pfeiffer et al 2006
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
MOLECULAR DIAGNOSTICS
bull PCR for detection of Aspergillus in blood BAL serum etc
bull Appear promising for rapid detection of Aspergillus directly on tissue or body fluids but
ndash False positive
ndash No standardized commercial method
bull Routine use of PCR can not be recommended
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
Mark Reinwald et al
bull ldquoTreatment with mould-active antifungals prior to BAL sampling significantly decreases the performance of the Aspergillus PCR assay in haematological patients if BAL was performed after administration of more than one antifungal agentrdquo
bull ldquoThe effect of antifungal therapy on other surrogate parameters suggests reduced sensitivity rates with antifungal therapy for GM whereas no influence of antifungal therapy on β-D-glucan performance in a (single-centre retrospective) study was describedrdquo
J Antimicrob Chemother 2012 67 2260ndash2267
CONCLUSION
bull Various detection methods of agents responsible for invasive fungal infections have been developed
bull The lack of sensitivity and specificity of the test uncertainty in diagnosing IFI decrease successful treatment
bull Need further studies to improve and develop laboratory diagnostic tool for IFI
LABORATORY DIAGNOSTIC OF IFI
bull (13)-β-D-Glucan assay accurate laboratory diagnostic tool in determining between definitive and probable invasive fungal infections
bull May be useful in the clinical practice if
ndash Implemented in a proper setting
ndash Interpreted after consideration of its limitation
Drosos E Karageorgopoulos et al Clinical Infectious Diseases 201152(6)750ndash770
bull The Glucatell serum BG detection assay is highly sensitive and specific as a diagnostic adjunct for IFI
Zekaver Odabasi et al
Clinical Infectious Diseases 2004 39199ndash205
LABORATORY DIAGNOSTIC OF IFI
INVASIVE CANDIDOSIS
SPECIES DISTRIBUTION
bull The incidence of Candidosis infection in hospitalized patients in the USA has increased (1980-1989) 8 responsible for all bloodstream infections
bull In the 1990s incidence of Candida bloodstream infections decreased due to a decline of C albicans infections however the incidence rate of C glabrata increased
bull 1992-1993 1998-2000 overall incidence of Candida bloodstream infections increased
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
bull Invasive candidosis
ndash endogenous
ndash transmission from person to person (direct patient-patient transmission healthcare workers medical devices contaminated intravenous infusions)
bull Cross infection in ICU
ndash C albicans via hand of healthcare workers
ndash C parapsilosis via medical devices
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
bull Invasive candidosis
ndash Neutropenic cancer patients
ndash Stem cellorgan transplants recipients
ndash ICU patients (adult surgical neonatal)
Widespread use of fluconazole emergence of less susceptible species
Note Intrinsic resistance (C glabrata and C krusei) to fluconazole factor in their emergence as pathogens
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
LABORATORY DIAGNOSTIC
bull Diagnosis of invasive candidosis is difficult due to a variation and non specific clinical presentation difficulties in intrepreting laboratory examination results
bull Dignosis based on clinical radiological microbiological and histopathological findings
bull Microbiology positive blood culture budding yeast cellspseudohyphae in specimens obtained from normal sterile or abnormal sites (biopsy aspiration)
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
LABORATORY DIAGNOSTIC
bull Investigation included
ndash Microscopy
ndash Culture
ndash Serological test
ndash D-arabinitol detection
ndash Molecular assays
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
MICROSCOPY
bull Typical budding yeast cells pseudohypahetrue hyphae of Candida species found in tissue biopsy normal sterile body fluids
bull C glabrata produces only yeast cells and
C albicans produces true hyphae in tissues
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
CULTURE
bull Isolation of Candida sp from sputum and faecal specimens can not be considered diagnostic of infection
bull Deep-seated infection Candida sp isolated from blood or normally sterile body fluids tissuesaspirates from other closed sites
bull Specimens should be examined as soon as possible after collection to avoid missinterpretation due to other organisms multiplication
bull Different susceptibility among Candida sp completed identification (speciation) before treatment
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
CULTURE
bull Blood cultures are positive in no more than 50 of neutropenic patients with dissemintaed candidosis or 80 of patients with endocarditis
bull Isolation of Candida sp from urine may indicate a serious infection
bull Isolation of Candida sp from LCS evidence for diagnosis of meningitis
bull Candida sp isolated from BAL in a patient with pulmonary infiltrates not sufficient for diagnosing candidosis
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
SEROLOGICAL TESTS
bull Limited usefulness
bull False positive (mucosal colonization superficial infection)
bull False negative (immunocompromised)
bull Antigen detection mannan insensitive
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
DETECTION OF D-ARABINITOL
bull D-arabinitol is produced by most of Candida sp (except C krusei and may be C glabrata)
bull It can be detected in serum or urine
bull Candidemia increased of serum D-arabiniitol-creatinine ratios has still to achieve widespread clinical use
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
MOLECULAR ASSAY
bull PCR detection of Candida DNA in blood serum LCS other specimens
bull Promising tools for rapid diagnosis of Candida sp from tissue or body-fluid
bull False positive
bull Standardized commercial assays were not available
bull Can not be recommended in diagnosis of invasive candidosis
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
The sources of yeast isolated
bull Sputum 49
bull Urine 13
bull Vaginal swab 9
bull Blood 8
bull Bronchial lavage 7
2010
bull Sputum 62
bull Vaginal swab 16
bull Blood 14
bull Urine 9
bull Bronchial lavage 5
2011
bull Sputum 102
bull Urine 18
bull Blood16 bull Bronchial lavage 12 bull Vaginal swab 11
2012
Candida spp isolated from blood
2010 2011 2012
C albicans 1 1 3
C parapsilosis 3 4 1
C pelliculosis 1 0 0
C lusitaniae 1 0 0
C tropicalis 0 2 10
C glabrata 0 1 0
C gulliermondii 0 1 1
Candida spp 1 0 0
7 9 15
Resistance pattern of Candida spp isolated from blood
2010 2011 2012
FCA ITR VRC FCA ITR VRC FCA ITR VRC
C albicans 17 115 115 115
C glabrata 19
C tropicalis 215 315 115
INVASIVE ASPERGILLOSIS
bull Incidence varies in ICU 58
bull In ICU risk factors including chronic obstructive pulmonary diseases and liver failure
bull Positive respiratory culture uncertain
bull Low sensitivity and specificity
bull Detection of galactomannan in BAL promising results
LABORATORY DIAGNOSTIC
bull Microscopy
bull Culture
bull Serological tests
bull Molecular diagnostics
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
MICROSCOPY
bull Histopathological examination of tissue biopsies immunohistochemical staining procedure
bull Non-pigmented septate hyphae dichotomous branching
bull Direct examination of sputum may help in diagnosing invasive aspergillosis BAL is better
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
A fumigatus
Richard C Barton Hindawi Publishing Corporation Scienti1048853 ca Volume 2013 Article ID 459405 29 pages httpdxdoiorg1011552013459405
CULTURE
bull Not sensitive interpreted with caution (contamination)
bull Recovered from blood LCS
bull Aspergillus isolated from BALsputum obtained from patients with a pulmonary infiltrates indicative of infection
bull A fumigatus can be isolated from sputum of patients suffer from allergic bronchopulmonary aspergillosis
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
SEROLOGICAL TESTS
bull Antobodies againts Aspergillus often helpful in the diagnosis of aspergillosis in non immunocompromised patients
bull Immunodiffusion indirect haemagglutination ELISA
bull The role is still uncertain
bull Positive ID test is not a proof of infection negative result can not exclude the diagnosis
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
SEROLOGICAL TESTS
bull Antigen detection (galactomannan) in blood and other body fluids rapid diagnosis of aspergilllosis in immunocompromised patients
bull Galactomannan fluctuate during the course of the infection useful if performed in a regular basis (twice per week)
bull Negative not exclude the diagnosis
bull ELISA for the detection of galactomannan has been approved for diagnostic use (the accuracy was moderate)
Christopher D Pfeiffer et al 2006
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
MOLECULAR DIAGNOSTICS
bull PCR for detection of Aspergillus in blood BAL serum etc
bull Appear promising for rapid detection of Aspergillus directly on tissue or body fluids but
ndash False positive
ndash No standardized commercial method
bull Routine use of PCR can not be recommended
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
Mark Reinwald et al
bull ldquoTreatment with mould-active antifungals prior to BAL sampling significantly decreases the performance of the Aspergillus PCR assay in haematological patients if BAL was performed after administration of more than one antifungal agentrdquo
bull ldquoThe effect of antifungal therapy on other surrogate parameters suggests reduced sensitivity rates with antifungal therapy for GM whereas no influence of antifungal therapy on β-D-glucan performance in a (single-centre retrospective) study was describedrdquo
J Antimicrob Chemother 2012 67 2260ndash2267
CONCLUSION
bull Various detection methods of agents responsible for invasive fungal infections have been developed
bull The lack of sensitivity and specificity of the test uncertainty in diagnosing IFI decrease successful treatment
bull Need further studies to improve and develop laboratory diagnostic tool for IFI
bull The Glucatell serum BG detection assay is highly sensitive and specific as a diagnostic adjunct for IFI
Zekaver Odabasi et al
Clinical Infectious Diseases 2004 39199ndash205
LABORATORY DIAGNOSTIC OF IFI
INVASIVE CANDIDOSIS
SPECIES DISTRIBUTION
bull The incidence of Candidosis infection in hospitalized patients in the USA has increased (1980-1989) 8 responsible for all bloodstream infections
bull In the 1990s incidence of Candida bloodstream infections decreased due to a decline of C albicans infections however the incidence rate of C glabrata increased
bull 1992-1993 1998-2000 overall incidence of Candida bloodstream infections increased
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
bull Invasive candidosis
ndash endogenous
ndash transmission from person to person (direct patient-patient transmission healthcare workers medical devices contaminated intravenous infusions)
bull Cross infection in ICU
ndash C albicans via hand of healthcare workers
ndash C parapsilosis via medical devices
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
bull Invasive candidosis
ndash Neutropenic cancer patients
ndash Stem cellorgan transplants recipients
ndash ICU patients (adult surgical neonatal)
Widespread use of fluconazole emergence of less susceptible species
Note Intrinsic resistance (C glabrata and C krusei) to fluconazole factor in their emergence as pathogens
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
LABORATORY DIAGNOSTIC
bull Diagnosis of invasive candidosis is difficult due to a variation and non specific clinical presentation difficulties in intrepreting laboratory examination results
bull Dignosis based on clinical radiological microbiological and histopathological findings
bull Microbiology positive blood culture budding yeast cellspseudohyphae in specimens obtained from normal sterile or abnormal sites (biopsy aspiration)
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
LABORATORY DIAGNOSTIC
bull Investigation included
ndash Microscopy
ndash Culture
ndash Serological test
ndash D-arabinitol detection
ndash Molecular assays
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
MICROSCOPY
bull Typical budding yeast cells pseudohypahetrue hyphae of Candida species found in tissue biopsy normal sterile body fluids
bull C glabrata produces only yeast cells and
C albicans produces true hyphae in tissues
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
CULTURE
bull Isolation of Candida sp from sputum and faecal specimens can not be considered diagnostic of infection
bull Deep-seated infection Candida sp isolated from blood or normally sterile body fluids tissuesaspirates from other closed sites
bull Specimens should be examined as soon as possible after collection to avoid missinterpretation due to other organisms multiplication
bull Different susceptibility among Candida sp completed identification (speciation) before treatment
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
CULTURE
bull Blood cultures are positive in no more than 50 of neutropenic patients with dissemintaed candidosis or 80 of patients with endocarditis
bull Isolation of Candida sp from urine may indicate a serious infection
bull Isolation of Candida sp from LCS evidence for diagnosis of meningitis
bull Candida sp isolated from BAL in a patient with pulmonary infiltrates not sufficient for diagnosing candidosis
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
SEROLOGICAL TESTS
bull Limited usefulness
bull False positive (mucosal colonization superficial infection)
bull False negative (immunocompromised)
bull Antigen detection mannan insensitive
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
DETECTION OF D-ARABINITOL
bull D-arabinitol is produced by most of Candida sp (except C krusei and may be C glabrata)
bull It can be detected in serum or urine
bull Candidemia increased of serum D-arabiniitol-creatinine ratios has still to achieve widespread clinical use
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
MOLECULAR ASSAY
bull PCR detection of Candida DNA in blood serum LCS other specimens
bull Promising tools for rapid diagnosis of Candida sp from tissue or body-fluid
bull False positive
bull Standardized commercial assays were not available
bull Can not be recommended in diagnosis of invasive candidosis
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
The sources of yeast isolated
bull Sputum 49
bull Urine 13
bull Vaginal swab 9
bull Blood 8
bull Bronchial lavage 7
2010
bull Sputum 62
bull Vaginal swab 16
bull Blood 14
bull Urine 9
bull Bronchial lavage 5
2011
bull Sputum 102
bull Urine 18
bull Blood16 bull Bronchial lavage 12 bull Vaginal swab 11
2012
Candida spp isolated from blood
2010 2011 2012
C albicans 1 1 3
C parapsilosis 3 4 1
C pelliculosis 1 0 0
C lusitaniae 1 0 0
C tropicalis 0 2 10
C glabrata 0 1 0
C gulliermondii 0 1 1
Candida spp 1 0 0
7 9 15
Resistance pattern of Candida spp isolated from blood
2010 2011 2012
FCA ITR VRC FCA ITR VRC FCA ITR VRC
C albicans 17 115 115 115
C glabrata 19
C tropicalis 215 315 115
INVASIVE ASPERGILLOSIS
bull Incidence varies in ICU 58
bull In ICU risk factors including chronic obstructive pulmonary diseases and liver failure
bull Positive respiratory culture uncertain
bull Low sensitivity and specificity
bull Detection of galactomannan in BAL promising results
LABORATORY DIAGNOSTIC
bull Microscopy
bull Culture
bull Serological tests
bull Molecular diagnostics
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
MICROSCOPY
bull Histopathological examination of tissue biopsies immunohistochemical staining procedure
bull Non-pigmented septate hyphae dichotomous branching
bull Direct examination of sputum may help in diagnosing invasive aspergillosis BAL is better
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
A fumigatus
Richard C Barton Hindawi Publishing Corporation Scienti1048853 ca Volume 2013 Article ID 459405 29 pages httpdxdoiorg1011552013459405
CULTURE
bull Not sensitive interpreted with caution (contamination)
bull Recovered from blood LCS
bull Aspergillus isolated from BALsputum obtained from patients with a pulmonary infiltrates indicative of infection
bull A fumigatus can be isolated from sputum of patients suffer from allergic bronchopulmonary aspergillosis
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
SEROLOGICAL TESTS
bull Antobodies againts Aspergillus often helpful in the diagnosis of aspergillosis in non immunocompromised patients
bull Immunodiffusion indirect haemagglutination ELISA
bull The role is still uncertain
bull Positive ID test is not a proof of infection negative result can not exclude the diagnosis
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
SEROLOGICAL TESTS
bull Antigen detection (galactomannan) in blood and other body fluids rapid diagnosis of aspergilllosis in immunocompromised patients
bull Galactomannan fluctuate during the course of the infection useful if performed in a regular basis (twice per week)
bull Negative not exclude the diagnosis
bull ELISA for the detection of galactomannan has been approved for diagnostic use (the accuracy was moderate)
Christopher D Pfeiffer et al 2006
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
MOLECULAR DIAGNOSTICS
bull PCR for detection of Aspergillus in blood BAL serum etc
bull Appear promising for rapid detection of Aspergillus directly on tissue or body fluids but
ndash False positive
ndash No standardized commercial method
bull Routine use of PCR can not be recommended
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
Mark Reinwald et al
bull ldquoTreatment with mould-active antifungals prior to BAL sampling significantly decreases the performance of the Aspergillus PCR assay in haematological patients if BAL was performed after administration of more than one antifungal agentrdquo
bull ldquoThe effect of antifungal therapy on other surrogate parameters suggests reduced sensitivity rates with antifungal therapy for GM whereas no influence of antifungal therapy on β-D-glucan performance in a (single-centre retrospective) study was describedrdquo
J Antimicrob Chemother 2012 67 2260ndash2267
CONCLUSION
bull Various detection methods of agents responsible for invasive fungal infections have been developed
bull The lack of sensitivity and specificity of the test uncertainty in diagnosing IFI decrease successful treatment
bull Need further studies to improve and develop laboratory diagnostic tool for IFI
INVASIVE CANDIDOSIS
SPECIES DISTRIBUTION
bull The incidence of Candidosis infection in hospitalized patients in the USA has increased (1980-1989) 8 responsible for all bloodstream infections
bull In the 1990s incidence of Candida bloodstream infections decreased due to a decline of C albicans infections however the incidence rate of C glabrata increased
bull 1992-1993 1998-2000 overall incidence of Candida bloodstream infections increased
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
bull Invasive candidosis
ndash endogenous
ndash transmission from person to person (direct patient-patient transmission healthcare workers medical devices contaminated intravenous infusions)
bull Cross infection in ICU
ndash C albicans via hand of healthcare workers
ndash C parapsilosis via medical devices
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
bull Invasive candidosis
ndash Neutropenic cancer patients
ndash Stem cellorgan transplants recipients
ndash ICU patients (adult surgical neonatal)
Widespread use of fluconazole emergence of less susceptible species
Note Intrinsic resistance (C glabrata and C krusei) to fluconazole factor in their emergence as pathogens
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
LABORATORY DIAGNOSTIC
bull Diagnosis of invasive candidosis is difficult due to a variation and non specific clinical presentation difficulties in intrepreting laboratory examination results
bull Dignosis based on clinical radiological microbiological and histopathological findings
bull Microbiology positive blood culture budding yeast cellspseudohyphae in specimens obtained from normal sterile or abnormal sites (biopsy aspiration)
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
LABORATORY DIAGNOSTIC
bull Investigation included
ndash Microscopy
ndash Culture
ndash Serological test
ndash D-arabinitol detection
ndash Molecular assays
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
MICROSCOPY
bull Typical budding yeast cells pseudohypahetrue hyphae of Candida species found in tissue biopsy normal sterile body fluids
bull C glabrata produces only yeast cells and
C albicans produces true hyphae in tissues
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
CULTURE
bull Isolation of Candida sp from sputum and faecal specimens can not be considered diagnostic of infection
bull Deep-seated infection Candida sp isolated from blood or normally sterile body fluids tissuesaspirates from other closed sites
bull Specimens should be examined as soon as possible after collection to avoid missinterpretation due to other organisms multiplication
bull Different susceptibility among Candida sp completed identification (speciation) before treatment
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
CULTURE
bull Blood cultures are positive in no more than 50 of neutropenic patients with dissemintaed candidosis or 80 of patients with endocarditis
bull Isolation of Candida sp from urine may indicate a serious infection
bull Isolation of Candida sp from LCS evidence for diagnosis of meningitis
bull Candida sp isolated from BAL in a patient with pulmonary infiltrates not sufficient for diagnosing candidosis
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
SEROLOGICAL TESTS
bull Limited usefulness
bull False positive (mucosal colonization superficial infection)
bull False negative (immunocompromised)
bull Antigen detection mannan insensitive
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
DETECTION OF D-ARABINITOL
bull D-arabinitol is produced by most of Candida sp (except C krusei and may be C glabrata)
bull It can be detected in serum or urine
bull Candidemia increased of serum D-arabiniitol-creatinine ratios has still to achieve widespread clinical use
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
MOLECULAR ASSAY
bull PCR detection of Candida DNA in blood serum LCS other specimens
bull Promising tools for rapid diagnosis of Candida sp from tissue or body-fluid
bull False positive
bull Standardized commercial assays were not available
bull Can not be recommended in diagnosis of invasive candidosis
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
The sources of yeast isolated
bull Sputum 49
bull Urine 13
bull Vaginal swab 9
bull Blood 8
bull Bronchial lavage 7
2010
bull Sputum 62
bull Vaginal swab 16
bull Blood 14
bull Urine 9
bull Bronchial lavage 5
2011
bull Sputum 102
bull Urine 18
bull Blood16 bull Bronchial lavage 12 bull Vaginal swab 11
2012
Candida spp isolated from blood
2010 2011 2012
C albicans 1 1 3
C parapsilosis 3 4 1
C pelliculosis 1 0 0
C lusitaniae 1 0 0
C tropicalis 0 2 10
C glabrata 0 1 0
C gulliermondii 0 1 1
Candida spp 1 0 0
7 9 15
Resistance pattern of Candida spp isolated from blood
2010 2011 2012
FCA ITR VRC FCA ITR VRC FCA ITR VRC
C albicans 17 115 115 115
C glabrata 19
C tropicalis 215 315 115
INVASIVE ASPERGILLOSIS
bull Incidence varies in ICU 58
bull In ICU risk factors including chronic obstructive pulmonary diseases and liver failure
bull Positive respiratory culture uncertain
bull Low sensitivity and specificity
bull Detection of galactomannan in BAL promising results
LABORATORY DIAGNOSTIC
bull Microscopy
bull Culture
bull Serological tests
bull Molecular diagnostics
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
MICROSCOPY
bull Histopathological examination of tissue biopsies immunohistochemical staining procedure
bull Non-pigmented septate hyphae dichotomous branching
bull Direct examination of sputum may help in diagnosing invasive aspergillosis BAL is better
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
A fumigatus
Richard C Barton Hindawi Publishing Corporation Scienti1048853 ca Volume 2013 Article ID 459405 29 pages httpdxdoiorg1011552013459405
CULTURE
bull Not sensitive interpreted with caution (contamination)
bull Recovered from blood LCS
bull Aspergillus isolated from BALsputum obtained from patients with a pulmonary infiltrates indicative of infection
bull A fumigatus can be isolated from sputum of patients suffer from allergic bronchopulmonary aspergillosis
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
SEROLOGICAL TESTS
bull Antobodies againts Aspergillus often helpful in the diagnosis of aspergillosis in non immunocompromised patients
bull Immunodiffusion indirect haemagglutination ELISA
bull The role is still uncertain
bull Positive ID test is not a proof of infection negative result can not exclude the diagnosis
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
SEROLOGICAL TESTS
bull Antigen detection (galactomannan) in blood and other body fluids rapid diagnosis of aspergilllosis in immunocompromised patients
bull Galactomannan fluctuate during the course of the infection useful if performed in a regular basis (twice per week)
bull Negative not exclude the diagnosis
bull ELISA for the detection of galactomannan has been approved for diagnostic use (the accuracy was moderate)
Christopher D Pfeiffer et al 2006
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
MOLECULAR DIAGNOSTICS
bull PCR for detection of Aspergillus in blood BAL serum etc
bull Appear promising for rapid detection of Aspergillus directly on tissue or body fluids but
ndash False positive
ndash No standardized commercial method
bull Routine use of PCR can not be recommended
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
Mark Reinwald et al
bull ldquoTreatment with mould-active antifungals prior to BAL sampling significantly decreases the performance of the Aspergillus PCR assay in haematological patients if BAL was performed after administration of more than one antifungal agentrdquo
bull ldquoThe effect of antifungal therapy on other surrogate parameters suggests reduced sensitivity rates with antifungal therapy for GM whereas no influence of antifungal therapy on β-D-glucan performance in a (single-centre retrospective) study was describedrdquo
J Antimicrob Chemother 2012 67 2260ndash2267
CONCLUSION
bull Various detection methods of agents responsible for invasive fungal infections have been developed
bull The lack of sensitivity and specificity of the test uncertainty in diagnosing IFI decrease successful treatment
bull Need further studies to improve and develop laboratory diagnostic tool for IFI
SPECIES DISTRIBUTION
bull The incidence of Candidosis infection in hospitalized patients in the USA has increased (1980-1989) 8 responsible for all bloodstream infections
bull In the 1990s incidence of Candida bloodstream infections decreased due to a decline of C albicans infections however the incidence rate of C glabrata increased
bull 1992-1993 1998-2000 overall incidence of Candida bloodstream infections increased
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
bull Invasive candidosis
ndash endogenous
ndash transmission from person to person (direct patient-patient transmission healthcare workers medical devices contaminated intravenous infusions)
bull Cross infection in ICU
ndash C albicans via hand of healthcare workers
ndash C parapsilosis via medical devices
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
bull Invasive candidosis
ndash Neutropenic cancer patients
ndash Stem cellorgan transplants recipients
ndash ICU patients (adult surgical neonatal)
Widespread use of fluconazole emergence of less susceptible species
Note Intrinsic resistance (C glabrata and C krusei) to fluconazole factor in their emergence as pathogens
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
LABORATORY DIAGNOSTIC
bull Diagnosis of invasive candidosis is difficult due to a variation and non specific clinical presentation difficulties in intrepreting laboratory examination results
bull Dignosis based on clinical radiological microbiological and histopathological findings
bull Microbiology positive blood culture budding yeast cellspseudohyphae in specimens obtained from normal sterile or abnormal sites (biopsy aspiration)
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
LABORATORY DIAGNOSTIC
bull Investigation included
ndash Microscopy
ndash Culture
ndash Serological test
ndash D-arabinitol detection
ndash Molecular assays
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
MICROSCOPY
bull Typical budding yeast cells pseudohypahetrue hyphae of Candida species found in tissue biopsy normal sterile body fluids
bull C glabrata produces only yeast cells and
C albicans produces true hyphae in tissues
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
CULTURE
bull Isolation of Candida sp from sputum and faecal specimens can not be considered diagnostic of infection
bull Deep-seated infection Candida sp isolated from blood or normally sterile body fluids tissuesaspirates from other closed sites
bull Specimens should be examined as soon as possible after collection to avoid missinterpretation due to other organisms multiplication
bull Different susceptibility among Candida sp completed identification (speciation) before treatment
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
CULTURE
bull Blood cultures are positive in no more than 50 of neutropenic patients with dissemintaed candidosis or 80 of patients with endocarditis
bull Isolation of Candida sp from urine may indicate a serious infection
bull Isolation of Candida sp from LCS evidence for diagnosis of meningitis
bull Candida sp isolated from BAL in a patient with pulmonary infiltrates not sufficient for diagnosing candidosis
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
SEROLOGICAL TESTS
bull Limited usefulness
bull False positive (mucosal colonization superficial infection)
bull False negative (immunocompromised)
bull Antigen detection mannan insensitive
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
DETECTION OF D-ARABINITOL
bull D-arabinitol is produced by most of Candida sp (except C krusei and may be C glabrata)
bull It can be detected in serum or urine
bull Candidemia increased of serum D-arabiniitol-creatinine ratios has still to achieve widespread clinical use
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
MOLECULAR ASSAY
bull PCR detection of Candida DNA in blood serum LCS other specimens
bull Promising tools for rapid diagnosis of Candida sp from tissue or body-fluid
bull False positive
bull Standardized commercial assays were not available
bull Can not be recommended in diagnosis of invasive candidosis
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
The sources of yeast isolated
bull Sputum 49
bull Urine 13
bull Vaginal swab 9
bull Blood 8
bull Bronchial lavage 7
2010
bull Sputum 62
bull Vaginal swab 16
bull Blood 14
bull Urine 9
bull Bronchial lavage 5
2011
bull Sputum 102
bull Urine 18
bull Blood16 bull Bronchial lavage 12 bull Vaginal swab 11
2012
Candida spp isolated from blood
2010 2011 2012
C albicans 1 1 3
C parapsilosis 3 4 1
C pelliculosis 1 0 0
C lusitaniae 1 0 0
C tropicalis 0 2 10
C glabrata 0 1 0
C gulliermondii 0 1 1
Candida spp 1 0 0
7 9 15
Resistance pattern of Candida spp isolated from blood
2010 2011 2012
FCA ITR VRC FCA ITR VRC FCA ITR VRC
C albicans 17 115 115 115
C glabrata 19
C tropicalis 215 315 115
INVASIVE ASPERGILLOSIS
bull Incidence varies in ICU 58
bull In ICU risk factors including chronic obstructive pulmonary diseases and liver failure
bull Positive respiratory culture uncertain
bull Low sensitivity and specificity
bull Detection of galactomannan in BAL promising results
LABORATORY DIAGNOSTIC
bull Microscopy
bull Culture
bull Serological tests
bull Molecular diagnostics
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
MICROSCOPY
bull Histopathological examination of tissue biopsies immunohistochemical staining procedure
bull Non-pigmented septate hyphae dichotomous branching
bull Direct examination of sputum may help in diagnosing invasive aspergillosis BAL is better
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
A fumigatus
Richard C Barton Hindawi Publishing Corporation Scienti1048853 ca Volume 2013 Article ID 459405 29 pages httpdxdoiorg1011552013459405
CULTURE
bull Not sensitive interpreted with caution (contamination)
bull Recovered from blood LCS
bull Aspergillus isolated from BALsputum obtained from patients with a pulmonary infiltrates indicative of infection
bull A fumigatus can be isolated from sputum of patients suffer from allergic bronchopulmonary aspergillosis
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
SEROLOGICAL TESTS
bull Antobodies againts Aspergillus often helpful in the diagnosis of aspergillosis in non immunocompromised patients
bull Immunodiffusion indirect haemagglutination ELISA
bull The role is still uncertain
bull Positive ID test is not a proof of infection negative result can not exclude the diagnosis
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
SEROLOGICAL TESTS
bull Antigen detection (galactomannan) in blood and other body fluids rapid diagnosis of aspergilllosis in immunocompromised patients
bull Galactomannan fluctuate during the course of the infection useful if performed in a regular basis (twice per week)
bull Negative not exclude the diagnosis
bull ELISA for the detection of galactomannan has been approved for diagnostic use (the accuracy was moderate)
Christopher D Pfeiffer et al 2006
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
MOLECULAR DIAGNOSTICS
bull PCR for detection of Aspergillus in blood BAL serum etc
bull Appear promising for rapid detection of Aspergillus directly on tissue or body fluids but
ndash False positive
ndash No standardized commercial method
bull Routine use of PCR can not be recommended
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
Mark Reinwald et al
bull ldquoTreatment with mould-active antifungals prior to BAL sampling significantly decreases the performance of the Aspergillus PCR assay in haematological patients if BAL was performed after administration of more than one antifungal agentrdquo
bull ldquoThe effect of antifungal therapy on other surrogate parameters suggests reduced sensitivity rates with antifungal therapy for GM whereas no influence of antifungal therapy on β-D-glucan performance in a (single-centre retrospective) study was describedrdquo
J Antimicrob Chemother 2012 67 2260ndash2267
CONCLUSION
bull Various detection methods of agents responsible for invasive fungal infections have been developed
bull The lack of sensitivity and specificity of the test uncertainty in diagnosing IFI decrease successful treatment
bull Need further studies to improve and develop laboratory diagnostic tool for IFI
bull The incidence of Candidosis infection in hospitalized patients in the USA has increased (1980-1989) 8 responsible for all bloodstream infections
bull In the 1990s incidence of Candida bloodstream infections decreased due to a decline of C albicans infections however the incidence rate of C glabrata increased
bull 1992-1993 1998-2000 overall incidence of Candida bloodstream infections increased
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
bull Invasive candidosis
ndash endogenous
ndash transmission from person to person (direct patient-patient transmission healthcare workers medical devices contaminated intravenous infusions)
bull Cross infection in ICU
ndash C albicans via hand of healthcare workers
ndash C parapsilosis via medical devices
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
bull Invasive candidosis
ndash Neutropenic cancer patients
ndash Stem cellorgan transplants recipients
ndash ICU patients (adult surgical neonatal)
Widespread use of fluconazole emergence of less susceptible species
Note Intrinsic resistance (C glabrata and C krusei) to fluconazole factor in their emergence as pathogens
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
LABORATORY DIAGNOSTIC
bull Diagnosis of invasive candidosis is difficult due to a variation and non specific clinical presentation difficulties in intrepreting laboratory examination results
bull Dignosis based on clinical radiological microbiological and histopathological findings
bull Microbiology positive blood culture budding yeast cellspseudohyphae in specimens obtained from normal sterile or abnormal sites (biopsy aspiration)
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
LABORATORY DIAGNOSTIC
bull Investigation included
ndash Microscopy
ndash Culture
ndash Serological test
ndash D-arabinitol detection
ndash Molecular assays
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
MICROSCOPY
bull Typical budding yeast cells pseudohypahetrue hyphae of Candida species found in tissue biopsy normal sterile body fluids
bull C glabrata produces only yeast cells and
C albicans produces true hyphae in tissues
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
CULTURE
bull Isolation of Candida sp from sputum and faecal specimens can not be considered diagnostic of infection
bull Deep-seated infection Candida sp isolated from blood or normally sterile body fluids tissuesaspirates from other closed sites
bull Specimens should be examined as soon as possible after collection to avoid missinterpretation due to other organisms multiplication
bull Different susceptibility among Candida sp completed identification (speciation) before treatment
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
CULTURE
bull Blood cultures are positive in no more than 50 of neutropenic patients with dissemintaed candidosis or 80 of patients with endocarditis
bull Isolation of Candida sp from urine may indicate a serious infection
bull Isolation of Candida sp from LCS evidence for diagnosis of meningitis
bull Candida sp isolated from BAL in a patient with pulmonary infiltrates not sufficient for diagnosing candidosis
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
SEROLOGICAL TESTS
bull Limited usefulness
bull False positive (mucosal colonization superficial infection)
bull False negative (immunocompromised)
bull Antigen detection mannan insensitive
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
DETECTION OF D-ARABINITOL
bull D-arabinitol is produced by most of Candida sp (except C krusei and may be C glabrata)
bull It can be detected in serum or urine
bull Candidemia increased of serum D-arabiniitol-creatinine ratios has still to achieve widespread clinical use
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
MOLECULAR ASSAY
bull PCR detection of Candida DNA in blood serum LCS other specimens
bull Promising tools for rapid diagnosis of Candida sp from tissue or body-fluid
bull False positive
bull Standardized commercial assays were not available
bull Can not be recommended in diagnosis of invasive candidosis
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
The sources of yeast isolated
bull Sputum 49
bull Urine 13
bull Vaginal swab 9
bull Blood 8
bull Bronchial lavage 7
2010
bull Sputum 62
bull Vaginal swab 16
bull Blood 14
bull Urine 9
bull Bronchial lavage 5
2011
bull Sputum 102
bull Urine 18
bull Blood16 bull Bronchial lavage 12 bull Vaginal swab 11
2012
Candida spp isolated from blood
2010 2011 2012
C albicans 1 1 3
C parapsilosis 3 4 1
C pelliculosis 1 0 0
C lusitaniae 1 0 0
C tropicalis 0 2 10
C glabrata 0 1 0
C gulliermondii 0 1 1
Candida spp 1 0 0
7 9 15
Resistance pattern of Candida spp isolated from blood
2010 2011 2012
FCA ITR VRC FCA ITR VRC FCA ITR VRC
C albicans 17 115 115 115
C glabrata 19
C tropicalis 215 315 115
INVASIVE ASPERGILLOSIS
bull Incidence varies in ICU 58
bull In ICU risk factors including chronic obstructive pulmonary diseases and liver failure
bull Positive respiratory culture uncertain
bull Low sensitivity and specificity
bull Detection of galactomannan in BAL promising results
LABORATORY DIAGNOSTIC
bull Microscopy
bull Culture
bull Serological tests
bull Molecular diagnostics
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
MICROSCOPY
bull Histopathological examination of tissue biopsies immunohistochemical staining procedure
bull Non-pigmented septate hyphae dichotomous branching
bull Direct examination of sputum may help in diagnosing invasive aspergillosis BAL is better
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
A fumigatus
Richard C Barton Hindawi Publishing Corporation Scienti1048853 ca Volume 2013 Article ID 459405 29 pages httpdxdoiorg1011552013459405
CULTURE
bull Not sensitive interpreted with caution (contamination)
bull Recovered from blood LCS
bull Aspergillus isolated from BALsputum obtained from patients with a pulmonary infiltrates indicative of infection
bull A fumigatus can be isolated from sputum of patients suffer from allergic bronchopulmonary aspergillosis
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
SEROLOGICAL TESTS
bull Antobodies againts Aspergillus often helpful in the diagnosis of aspergillosis in non immunocompromised patients
bull Immunodiffusion indirect haemagglutination ELISA
bull The role is still uncertain
bull Positive ID test is not a proof of infection negative result can not exclude the diagnosis
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
SEROLOGICAL TESTS
bull Antigen detection (galactomannan) in blood and other body fluids rapid diagnosis of aspergilllosis in immunocompromised patients
bull Galactomannan fluctuate during the course of the infection useful if performed in a regular basis (twice per week)
bull Negative not exclude the diagnosis
bull ELISA for the detection of galactomannan has been approved for diagnostic use (the accuracy was moderate)
Christopher D Pfeiffer et al 2006
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
MOLECULAR DIAGNOSTICS
bull PCR for detection of Aspergillus in blood BAL serum etc
bull Appear promising for rapid detection of Aspergillus directly on tissue or body fluids but
ndash False positive
ndash No standardized commercial method
bull Routine use of PCR can not be recommended
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
Mark Reinwald et al
bull ldquoTreatment with mould-active antifungals prior to BAL sampling significantly decreases the performance of the Aspergillus PCR assay in haematological patients if BAL was performed after administration of more than one antifungal agentrdquo
bull ldquoThe effect of antifungal therapy on other surrogate parameters suggests reduced sensitivity rates with antifungal therapy for GM whereas no influence of antifungal therapy on β-D-glucan performance in a (single-centre retrospective) study was describedrdquo
J Antimicrob Chemother 2012 67 2260ndash2267
CONCLUSION
bull Various detection methods of agents responsible for invasive fungal infections have been developed
bull The lack of sensitivity and specificity of the test uncertainty in diagnosing IFI decrease successful treatment
bull Need further studies to improve and develop laboratory diagnostic tool for IFI
bull Invasive candidosis
ndash endogenous
ndash transmission from person to person (direct patient-patient transmission healthcare workers medical devices contaminated intravenous infusions)
bull Cross infection in ICU
ndash C albicans via hand of healthcare workers
ndash C parapsilosis via medical devices
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
bull Invasive candidosis
ndash Neutropenic cancer patients
ndash Stem cellorgan transplants recipients
ndash ICU patients (adult surgical neonatal)
Widespread use of fluconazole emergence of less susceptible species
Note Intrinsic resistance (C glabrata and C krusei) to fluconazole factor in their emergence as pathogens
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
LABORATORY DIAGNOSTIC
bull Diagnosis of invasive candidosis is difficult due to a variation and non specific clinical presentation difficulties in intrepreting laboratory examination results
bull Dignosis based on clinical radiological microbiological and histopathological findings
bull Microbiology positive blood culture budding yeast cellspseudohyphae in specimens obtained from normal sterile or abnormal sites (biopsy aspiration)
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
LABORATORY DIAGNOSTIC
bull Investigation included
ndash Microscopy
ndash Culture
ndash Serological test
ndash D-arabinitol detection
ndash Molecular assays
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
MICROSCOPY
bull Typical budding yeast cells pseudohypahetrue hyphae of Candida species found in tissue biopsy normal sterile body fluids
bull C glabrata produces only yeast cells and
C albicans produces true hyphae in tissues
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
CULTURE
bull Isolation of Candida sp from sputum and faecal specimens can not be considered diagnostic of infection
bull Deep-seated infection Candida sp isolated from blood or normally sterile body fluids tissuesaspirates from other closed sites
bull Specimens should be examined as soon as possible after collection to avoid missinterpretation due to other organisms multiplication
bull Different susceptibility among Candida sp completed identification (speciation) before treatment
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
CULTURE
bull Blood cultures are positive in no more than 50 of neutropenic patients with dissemintaed candidosis or 80 of patients with endocarditis
bull Isolation of Candida sp from urine may indicate a serious infection
bull Isolation of Candida sp from LCS evidence for diagnosis of meningitis
bull Candida sp isolated from BAL in a patient with pulmonary infiltrates not sufficient for diagnosing candidosis
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
SEROLOGICAL TESTS
bull Limited usefulness
bull False positive (mucosal colonization superficial infection)
bull False negative (immunocompromised)
bull Antigen detection mannan insensitive
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
DETECTION OF D-ARABINITOL
bull D-arabinitol is produced by most of Candida sp (except C krusei and may be C glabrata)
bull It can be detected in serum or urine
bull Candidemia increased of serum D-arabiniitol-creatinine ratios has still to achieve widespread clinical use
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
MOLECULAR ASSAY
bull PCR detection of Candida DNA in blood serum LCS other specimens
bull Promising tools for rapid diagnosis of Candida sp from tissue or body-fluid
bull False positive
bull Standardized commercial assays were not available
bull Can not be recommended in diagnosis of invasive candidosis
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
The sources of yeast isolated
bull Sputum 49
bull Urine 13
bull Vaginal swab 9
bull Blood 8
bull Bronchial lavage 7
2010
bull Sputum 62
bull Vaginal swab 16
bull Blood 14
bull Urine 9
bull Bronchial lavage 5
2011
bull Sputum 102
bull Urine 18
bull Blood16 bull Bronchial lavage 12 bull Vaginal swab 11
2012
Candida spp isolated from blood
2010 2011 2012
C albicans 1 1 3
C parapsilosis 3 4 1
C pelliculosis 1 0 0
C lusitaniae 1 0 0
C tropicalis 0 2 10
C glabrata 0 1 0
C gulliermondii 0 1 1
Candida spp 1 0 0
7 9 15
Resistance pattern of Candida spp isolated from blood
2010 2011 2012
FCA ITR VRC FCA ITR VRC FCA ITR VRC
C albicans 17 115 115 115
C glabrata 19
C tropicalis 215 315 115
INVASIVE ASPERGILLOSIS
bull Incidence varies in ICU 58
bull In ICU risk factors including chronic obstructive pulmonary diseases and liver failure
bull Positive respiratory culture uncertain
bull Low sensitivity and specificity
bull Detection of galactomannan in BAL promising results
LABORATORY DIAGNOSTIC
bull Microscopy
bull Culture
bull Serological tests
bull Molecular diagnostics
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
MICROSCOPY
bull Histopathological examination of tissue biopsies immunohistochemical staining procedure
bull Non-pigmented septate hyphae dichotomous branching
bull Direct examination of sputum may help in diagnosing invasive aspergillosis BAL is better
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
A fumigatus
Richard C Barton Hindawi Publishing Corporation Scienti1048853 ca Volume 2013 Article ID 459405 29 pages httpdxdoiorg1011552013459405
CULTURE
bull Not sensitive interpreted with caution (contamination)
bull Recovered from blood LCS
bull Aspergillus isolated from BALsputum obtained from patients with a pulmonary infiltrates indicative of infection
bull A fumigatus can be isolated from sputum of patients suffer from allergic bronchopulmonary aspergillosis
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
SEROLOGICAL TESTS
bull Antobodies againts Aspergillus often helpful in the diagnosis of aspergillosis in non immunocompromised patients
bull Immunodiffusion indirect haemagglutination ELISA
bull The role is still uncertain
bull Positive ID test is not a proof of infection negative result can not exclude the diagnosis
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
SEROLOGICAL TESTS
bull Antigen detection (galactomannan) in blood and other body fluids rapid diagnosis of aspergilllosis in immunocompromised patients
bull Galactomannan fluctuate during the course of the infection useful if performed in a regular basis (twice per week)
bull Negative not exclude the diagnosis
bull ELISA for the detection of galactomannan has been approved for diagnostic use (the accuracy was moderate)
Christopher D Pfeiffer et al 2006
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
MOLECULAR DIAGNOSTICS
bull PCR for detection of Aspergillus in blood BAL serum etc
bull Appear promising for rapid detection of Aspergillus directly on tissue or body fluids but
ndash False positive
ndash No standardized commercial method
bull Routine use of PCR can not be recommended
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
Mark Reinwald et al
bull ldquoTreatment with mould-active antifungals prior to BAL sampling significantly decreases the performance of the Aspergillus PCR assay in haematological patients if BAL was performed after administration of more than one antifungal agentrdquo
bull ldquoThe effect of antifungal therapy on other surrogate parameters suggests reduced sensitivity rates with antifungal therapy for GM whereas no influence of antifungal therapy on β-D-glucan performance in a (single-centre retrospective) study was describedrdquo
J Antimicrob Chemother 2012 67 2260ndash2267
CONCLUSION
bull Various detection methods of agents responsible for invasive fungal infections have been developed
bull The lack of sensitivity and specificity of the test uncertainty in diagnosing IFI decrease successful treatment
bull Need further studies to improve and develop laboratory diagnostic tool for IFI
bull Invasive candidosis
ndash Neutropenic cancer patients
ndash Stem cellorgan transplants recipients
ndash ICU patients (adult surgical neonatal)
Widespread use of fluconazole emergence of less susceptible species
Note Intrinsic resistance (C glabrata and C krusei) to fluconazole factor in their emergence as pathogens
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
LABORATORY DIAGNOSTIC
bull Diagnosis of invasive candidosis is difficult due to a variation and non specific clinical presentation difficulties in intrepreting laboratory examination results
bull Dignosis based on clinical radiological microbiological and histopathological findings
bull Microbiology positive blood culture budding yeast cellspseudohyphae in specimens obtained from normal sterile or abnormal sites (biopsy aspiration)
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
LABORATORY DIAGNOSTIC
bull Investigation included
ndash Microscopy
ndash Culture
ndash Serological test
ndash D-arabinitol detection
ndash Molecular assays
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
MICROSCOPY
bull Typical budding yeast cells pseudohypahetrue hyphae of Candida species found in tissue biopsy normal sterile body fluids
bull C glabrata produces only yeast cells and
C albicans produces true hyphae in tissues
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
CULTURE
bull Isolation of Candida sp from sputum and faecal specimens can not be considered diagnostic of infection
bull Deep-seated infection Candida sp isolated from blood or normally sterile body fluids tissuesaspirates from other closed sites
bull Specimens should be examined as soon as possible after collection to avoid missinterpretation due to other organisms multiplication
bull Different susceptibility among Candida sp completed identification (speciation) before treatment
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
CULTURE
bull Blood cultures are positive in no more than 50 of neutropenic patients with dissemintaed candidosis or 80 of patients with endocarditis
bull Isolation of Candida sp from urine may indicate a serious infection
bull Isolation of Candida sp from LCS evidence for diagnosis of meningitis
bull Candida sp isolated from BAL in a patient with pulmonary infiltrates not sufficient for diagnosing candidosis
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
SEROLOGICAL TESTS
bull Limited usefulness
bull False positive (mucosal colonization superficial infection)
bull False negative (immunocompromised)
bull Antigen detection mannan insensitive
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
DETECTION OF D-ARABINITOL
bull D-arabinitol is produced by most of Candida sp (except C krusei and may be C glabrata)
bull It can be detected in serum or urine
bull Candidemia increased of serum D-arabiniitol-creatinine ratios has still to achieve widespread clinical use
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
MOLECULAR ASSAY
bull PCR detection of Candida DNA in blood serum LCS other specimens
bull Promising tools for rapid diagnosis of Candida sp from tissue or body-fluid
bull False positive
bull Standardized commercial assays were not available
bull Can not be recommended in diagnosis of invasive candidosis
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
The sources of yeast isolated
bull Sputum 49
bull Urine 13
bull Vaginal swab 9
bull Blood 8
bull Bronchial lavage 7
2010
bull Sputum 62
bull Vaginal swab 16
bull Blood 14
bull Urine 9
bull Bronchial lavage 5
2011
bull Sputum 102
bull Urine 18
bull Blood16 bull Bronchial lavage 12 bull Vaginal swab 11
2012
Candida spp isolated from blood
2010 2011 2012
C albicans 1 1 3
C parapsilosis 3 4 1
C pelliculosis 1 0 0
C lusitaniae 1 0 0
C tropicalis 0 2 10
C glabrata 0 1 0
C gulliermondii 0 1 1
Candida spp 1 0 0
7 9 15
Resistance pattern of Candida spp isolated from blood
2010 2011 2012
FCA ITR VRC FCA ITR VRC FCA ITR VRC
C albicans 17 115 115 115
C glabrata 19
C tropicalis 215 315 115
INVASIVE ASPERGILLOSIS
bull Incidence varies in ICU 58
bull In ICU risk factors including chronic obstructive pulmonary diseases and liver failure
bull Positive respiratory culture uncertain
bull Low sensitivity and specificity
bull Detection of galactomannan in BAL promising results
LABORATORY DIAGNOSTIC
bull Microscopy
bull Culture
bull Serological tests
bull Molecular diagnostics
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
MICROSCOPY
bull Histopathological examination of tissue biopsies immunohistochemical staining procedure
bull Non-pigmented septate hyphae dichotomous branching
bull Direct examination of sputum may help in diagnosing invasive aspergillosis BAL is better
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
A fumigatus
Richard C Barton Hindawi Publishing Corporation Scienti1048853 ca Volume 2013 Article ID 459405 29 pages httpdxdoiorg1011552013459405
CULTURE
bull Not sensitive interpreted with caution (contamination)
bull Recovered from blood LCS
bull Aspergillus isolated from BALsputum obtained from patients with a pulmonary infiltrates indicative of infection
bull A fumigatus can be isolated from sputum of patients suffer from allergic bronchopulmonary aspergillosis
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
SEROLOGICAL TESTS
bull Antobodies againts Aspergillus often helpful in the diagnosis of aspergillosis in non immunocompromised patients
bull Immunodiffusion indirect haemagglutination ELISA
bull The role is still uncertain
bull Positive ID test is not a proof of infection negative result can not exclude the diagnosis
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
SEROLOGICAL TESTS
bull Antigen detection (galactomannan) in blood and other body fluids rapid diagnosis of aspergilllosis in immunocompromised patients
bull Galactomannan fluctuate during the course of the infection useful if performed in a regular basis (twice per week)
bull Negative not exclude the diagnosis
bull ELISA for the detection of galactomannan has been approved for diagnostic use (the accuracy was moderate)
Christopher D Pfeiffer et al 2006
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
MOLECULAR DIAGNOSTICS
bull PCR for detection of Aspergillus in blood BAL serum etc
bull Appear promising for rapid detection of Aspergillus directly on tissue or body fluids but
ndash False positive
ndash No standardized commercial method
bull Routine use of PCR can not be recommended
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
Mark Reinwald et al
bull ldquoTreatment with mould-active antifungals prior to BAL sampling significantly decreases the performance of the Aspergillus PCR assay in haematological patients if BAL was performed after administration of more than one antifungal agentrdquo
bull ldquoThe effect of antifungal therapy on other surrogate parameters suggests reduced sensitivity rates with antifungal therapy for GM whereas no influence of antifungal therapy on β-D-glucan performance in a (single-centre retrospective) study was describedrdquo
J Antimicrob Chemother 2012 67 2260ndash2267
CONCLUSION
bull Various detection methods of agents responsible for invasive fungal infections have been developed
bull The lack of sensitivity and specificity of the test uncertainty in diagnosing IFI decrease successful treatment
bull Need further studies to improve and develop laboratory diagnostic tool for IFI
LABORATORY DIAGNOSTIC
bull Diagnosis of invasive candidosis is difficult due to a variation and non specific clinical presentation difficulties in intrepreting laboratory examination results
bull Dignosis based on clinical radiological microbiological and histopathological findings
bull Microbiology positive blood culture budding yeast cellspseudohyphae in specimens obtained from normal sterile or abnormal sites (biopsy aspiration)
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
LABORATORY DIAGNOSTIC
bull Investigation included
ndash Microscopy
ndash Culture
ndash Serological test
ndash D-arabinitol detection
ndash Molecular assays
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
MICROSCOPY
bull Typical budding yeast cells pseudohypahetrue hyphae of Candida species found in tissue biopsy normal sterile body fluids
bull C glabrata produces only yeast cells and
C albicans produces true hyphae in tissues
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
CULTURE
bull Isolation of Candida sp from sputum and faecal specimens can not be considered diagnostic of infection
bull Deep-seated infection Candida sp isolated from blood or normally sterile body fluids tissuesaspirates from other closed sites
bull Specimens should be examined as soon as possible after collection to avoid missinterpretation due to other organisms multiplication
bull Different susceptibility among Candida sp completed identification (speciation) before treatment
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
CULTURE
bull Blood cultures are positive in no more than 50 of neutropenic patients with dissemintaed candidosis or 80 of patients with endocarditis
bull Isolation of Candida sp from urine may indicate a serious infection
bull Isolation of Candida sp from LCS evidence for diagnosis of meningitis
bull Candida sp isolated from BAL in a patient with pulmonary infiltrates not sufficient for diagnosing candidosis
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
SEROLOGICAL TESTS
bull Limited usefulness
bull False positive (mucosal colonization superficial infection)
bull False negative (immunocompromised)
bull Antigen detection mannan insensitive
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
DETECTION OF D-ARABINITOL
bull D-arabinitol is produced by most of Candida sp (except C krusei and may be C glabrata)
bull It can be detected in serum or urine
bull Candidemia increased of serum D-arabiniitol-creatinine ratios has still to achieve widespread clinical use
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
MOLECULAR ASSAY
bull PCR detection of Candida DNA in blood serum LCS other specimens
bull Promising tools for rapid diagnosis of Candida sp from tissue or body-fluid
bull False positive
bull Standardized commercial assays were not available
bull Can not be recommended in diagnosis of invasive candidosis
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
The sources of yeast isolated
bull Sputum 49
bull Urine 13
bull Vaginal swab 9
bull Blood 8
bull Bronchial lavage 7
2010
bull Sputum 62
bull Vaginal swab 16
bull Blood 14
bull Urine 9
bull Bronchial lavage 5
2011
bull Sputum 102
bull Urine 18
bull Blood16 bull Bronchial lavage 12 bull Vaginal swab 11
2012
Candida spp isolated from blood
2010 2011 2012
C albicans 1 1 3
C parapsilosis 3 4 1
C pelliculosis 1 0 0
C lusitaniae 1 0 0
C tropicalis 0 2 10
C glabrata 0 1 0
C gulliermondii 0 1 1
Candida spp 1 0 0
7 9 15
Resistance pattern of Candida spp isolated from blood
2010 2011 2012
FCA ITR VRC FCA ITR VRC FCA ITR VRC
C albicans 17 115 115 115
C glabrata 19
C tropicalis 215 315 115
INVASIVE ASPERGILLOSIS
bull Incidence varies in ICU 58
bull In ICU risk factors including chronic obstructive pulmonary diseases and liver failure
bull Positive respiratory culture uncertain
bull Low sensitivity and specificity
bull Detection of galactomannan in BAL promising results
LABORATORY DIAGNOSTIC
bull Microscopy
bull Culture
bull Serological tests
bull Molecular diagnostics
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
MICROSCOPY
bull Histopathological examination of tissue biopsies immunohistochemical staining procedure
bull Non-pigmented septate hyphae dichotomous branching
bull Direct examination of sputum may help in diagnosing invasive aspergillosis BAL is better
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
A fumigatus
Richard C Barton Hindawi Publishing Corporation Scienti1048853 ca Volume 2013 Article ID 459405 29 pages httpdxdoiorg1011552013459405
CULTURE
bull Not sensitive interpreted with caution (contamination)
bull Recovered from blood LCS
bull Aspergillus isolated from BALsputum obtained from patients with a pulmonary infiltrates indicative of infection
bull A fumigatus can be isolated from sputum of patients suffer from allergic bronchopulmonary aspergillosis
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
SEROLOGICAL TESTS
bull Antobodies againts Aspergillus often helpful in the diagnosis of aspergillosis in non immunocompromised patients
bull Immunodiffusion indirect haemagglutination ELISA
bull The role is still uncertain
bull Positive ID test is not a proof of infection negative result can not exclude the diagnosis
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
SEROLOGICAL TESTS
bull Antigen detection (galactomannan) in blood and other body fluids rapid diagnosis of aspergilllosis in immunocompromised patients
bull Galactomannan fluctuate during the course of the infection useful if performed in a regular basis (twice per week)
bull Negative not exclude the diagnosis
bull ELISA for the detection of galactomannan has been approved for diagnostic use (the accuracy was moderate)
Christopher D Pfeiffer et al 2006
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
MOLECULAR DIAGNOSTICS
bull PCR for detection of Aspergillus in blood BAL serum etc
bull Appear promising for rapid detection of Aspergillus directly on tissue or body fluids but
ndash False positive
ndash No standardized commercial method
bull Routine use of PCR can not be recommended
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
Mark Reinwald et al
bull ldquoTreatment with mould-active antifungals prior to BAL sampling significantly decreases the performance of the Aspergillus PCR assay in haematological patients if BAL was performed after administration of more than one antifungal agentrdquo
bull ldquoThe effect of antifungal therapy on other surrogate parameters suggests reduced sensitivity rates with antifungal therapy for GM whereas no influence of antifungal therapy on β-D-glucan performance in a (single-centre retrospective) study was describedrdquo
J Antimicrob Chemother 2012 67 2260ndash2267
CONCLUSION
bull Various detection methods of agents responsible for invasive fungal infections have been developed
bull The lack of sensitivity and specificity of the test uncertainty in diagnosing IFI decrease successful treatment
bull Need further studies to improve and develop laboratory diagnostic tool for IFI
LABORATORY DIAGNOSTIC
bull Investigation included
ndash Microscopy
ndash Culture
ndash Serological test
ndash D-arabinitol detection
ndash Molecular assays
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
MICROSCOPY
bull Typical budding yeast cells pseudohypahetrue hyphae of Candida species found in tissue biopsy normal sterile body fluids
bull C glabrata produces only yeast cells and
C albicans produces true hyphae in tissues
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
CULTURE
bull Isolation of Candida sp from sputum and faecal specimens can not be considered diagnostic of infection
bull Deep-seated infection Candida sp isolated from blood or normally sterile body fluids tissuesaspirates from other closed sites
bull Specimens should be examined as soon as possible after collection to avoid missinterpretation due to other organisms multiplication
bull Different susceptibility among Candida sp completed identification (speciation) before treatment
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
CULTURE
bull Blood cultures are positive in no more than 50 of neutropenic patients with dissemintaed candidosis or 80 of patients with endocarditis
bull Isolation of Candida sp from urine may indicate a serious infection
bull Isolation of Candida sp from LCS evidence for diagnosis of meningitis
bull Candida sp isolated from BAL in a patient with pulmonary infiltrates not sufficient for diagnosing candidosis
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
SEROLOGICAL TESTS
bull Limited usefulness
bull False positive (mucosal colonization superficial infection)
bull False negative (immunocompromised)
bull Antigen detection mannan insensitive
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
DETECTION OF D-ARABINITOL
bull D-arabinitol is produced by most of Candida sp (except C krusei and may be C glabrata)
bull It can be detected in serum or urine
bull Candidemia increased of serum D-arabiniitol-creatinine ratios has still to achieve widespread clinical use
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
MOLECULAR ASSAY
bull PCR detection of Candida DNA in blood serum LCS other specimens
bull Promising tools for rapid diagnosis of Candida sp from tissue or body-fluid
bull False positive
bull Standardized commercial assays were not available
bull Can not be recommended in diagnosis of invasive candidosis
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
The sources of yeast isolated
bull Sputum 49
bull Urine 13
bull Vaginal swab 9
bull Blood 8
bull Bronchial lavage 7
2010
bull Sputum 62
bull Vaginal swab 16
bull Blood 14
bull Urine 9
bull Bronchial lavage 5
2011
bull Sputum 102
bull Urine 18
bull Blood16 bull Bronchial lavage 12 bull Vaginal swab 11
2012
Candida spp isolated from blood
2010 2011 2012
C albicans 1 1 3
C parapsilosis 3 4 1
C pelliculosis 1 0 0
C lusitaniae 1 0 0
C tropicalis 0 2 10
C glabrata 0 1 0
C gulliermondii 0 1 1
Candida spp 1 0 0
7 9 15
Resistance pattern of Candida spp isolated from blood
2010 2011 2012
FCA ITR VRC FCA ITR VRC FCA ITR VRC
C albicans 17 115 115 115
C glabrata 19
C tropicalis 215 315 115
INVASIVE ASPERGILLOSIS
bull Incidence varies in ICU 58
bull In ICU risk factors including chronic obstructive pulmonary diseases and liver failure
bull Positive respiratory culture uncertain
bull Low sensitivity and specificity
bull Detection of galactomannan in BAL promising results
LABORATORY DIAGNOSTIC
bull Microscopy
bull Culture
bull Serological tests
bull Molecular diagnostics
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
MICROSCOPY
bull Histopathological examination of tissue biopsies immunohistochemical staining procedure
bull Non-pigmented septate hyphae dichotomous branching
bull Direct examination of sputum may help in diagnosing invasive aspergillosis BAL is better
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
A fumigatus
Richard C Barton Hindawi Publishing Corporation Scienti1048853 ca Volume 2013 Article ID 459405 29 pages httpdxdoiorg1011552013459405
CULTURE
bull Not sensitive interpreted with caution (contamination)
bull Recovered from blood LCS
bull Aspergillus isolated from BALsputum obtained from patients with a pulmonary infiltrates indicative of infection
bull A fumigatus can be isolated from sputum of patients suffer from allergic bronchopulmonary aspergillosis
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
SEROLOGICAL TESTS
bull Antobodies againts Aspergillus often helpful in the diagnosis of aspergillosis in non immunocompromised patients
bull Immunodiffusion indirect haemagglutination ELISA
bull The role is still uncertain
bull Positive ID test is not a proof of infection negative result can not exclude the diagnosis
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
SEROLOGICAL TESTS
bull Antigen detection (galactomannan) in blood and other body fluids rapid diagnosis of aspergilllosis in immunocompromised patients
bull Galactomannan fluctuate during the course of the infection useful if performed in a regular basis (twice per week)
bull Negative not exclude the diagnosis
bull ELISA for the detection of galactomannan has been approved for diagnostic use (the accuracy was moderate)
Christopher D Pfeiffer et al 2006
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
MOLECULAR DIAGNOSTICS
bull PCR for detection of Aspergillus in blood BAL serum etc
bull Appear promising for rapid detection of Aspergillus directly on tissue or body fluids but
ndash False positive
ndash No standardized commercial method
bull Routine use of PCR can not be recommended
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
Mark Reinwald et al
bull ldquoTreatment with mould-active antifungals prior to BAL sampling significantly decreases the performance of the Aspergillus PCR assay in haematological patients if BAL was performed after administration of more than one antifungal agentrdquo
bull ldquoThe effect of antifungal therapy on other surrogate parameters suggests reduced sensitivity rates with antifungal therapy for GM whereas no influence of antifungal therapy on β-D-glucan performance in a (single-centre retrospective) study was describedrdquo
J Antimicrob Chemother 2012 67 2260ndash2267
CONCLUSION
bull Various detection methods of agents responsible for invasive fungal infections have been developed
bull The lack of sensitivity and specificity of the test uncertainty in diagnosing IFI decrease successful treatment
bull Need further studies to improve and develop laboratory diagnostic tool for IFI
MICROSCOPY
bull Typical budding yeast cells pseudohypahetrue hyphae of Candida species found in tissue biopsy normal sterile body fluids
bull C glabrata produces only yeast cells and
C albicans produces true hyphae in tissues
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
CULTURE
bull Isolation of Candida sp from sputum and faecal specimens can not be considered diagnostic of infection
bull Deep-seated infection Candida sp isolated from blood or normally sterile body fluids tissuesaspirates from other closed sites
bull Specimens should be examined as soon as possible after collection to avoid missinterpretation due to other organisms multiplication
bull Different susceptibility among Candida sp completed identification (speciation) before treatment
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
CULTURE
bull Blood cultures are positive in no more than 50 of neutropenic patients with dissemintaed candidosis or 80 of patients with endocarditis
bull Isolation of Candida sp from urine may indicate a serious infection
bull Isolation of Candida sp from LCS evidence for diagnosis of meningitis
bull Candida sp isolated from BAL in a patient with pulmonary infiltrates not sufficient for diagnosing candidosis
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
SEROLOGICAL TESTS
bull Limited usefulness
bull False positive (mucosal colonization superficial infection)
bull False negative (immunocompromised)
bull Antigen detection mannan insensitive
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
DETECTION OF D-ARABINITOL
bull D-arabinitol is produced by most of Candida sp (except C krusei and may be C glabrata)
bull It can be detected in serum or urine
bull Candidemia increased of serum D-arabiniitol-creatinine ratios has still to achieve widespread clinical use
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
MOLECULAR ASSAY
bull PCR detection of Candida DNA in blood serum LCS other specimens
bull Promising tools for rapid diagnosis of Candida sp from tissue or body-fluid
bull False positive
bull Standardized commercial assays were not available
bull Can not be recommended in diagnosis of invasive candidosis
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
The sources of yeast isolated
bull Sputum 49
bull Urine 13
bull Vaginal swab 9
bull Blood 8
bull Bronchial lavage 7
2010
bull Sputum 62
bull Vaginal swab 16
bull Blood 14
bull Urine 9
bull Bronchial lavage 5
2011
bull Sputum 102
bull Urine 18
bull Blood16 bull Bronchial lavage 12 bull Vaginal swab 11
2012
Candida spp isolated from blood
2010 2011 2012
C albicans 1 1 3
C parapsilosis 3 4 1
C pelliculosis 1 0 0
C lusitaniae 1 0 0
C tropicalis 0 2 10
C glabrata 0 1 0
C gulliermondii 0 1 1
Candida spp 1 0 0
7 9 15
Resistance pattern of Candida spp isolated from blood
2010 2011 2012
FCA ITR VRC FCA ITR VRC FCA ITR VRC
C albicans 17 115 115 115
C glabrata 19
C tropicalis 215 315 115
INVASIVE ASPERGILLOSIS
bull Incidence varies in ICU 58
bull In ICU risk factors including chronic obstructive pulmonary diseases and liver failure
bull Positive respiratory culture uncertain
bull Low sensitivity and specificity
bull Detection of galactomannan in BAL promising results
LABORATORY DIAGNOSTIC
bull Microscopy
bull Culture
bull Serological tests
bull Molecular diagnostics
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
MICROSCOPY
bull Histopathological examination of tissue biopsies immunohistochemical staining procedure
bull Non-pigmented septate hyphae dichotomous branching
bull Direct examination of sputum may help in diagnosing invasive aspergillosis BAL is better
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
A fumigatus
Richard C Barton Hindawi Publishing Corporation Scienti1048853 ca Volume 2013 Article ID 459405 29 pages httpdxdoiorg1011552013459405
CULTURE
bull Not sensitive interpreted with caution (contamination)
bull Recovered from blood LCS
bull Aspergillus isolated from BALsputum obtained from patients with a pulmonary infiltrates indicative of infection
bull A fumigatus can be isolated from sputum of patients suffer from allergic bronchopulmonary aspergillosis
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
SEROLOGICAL TESTS
bull Antobodies againts Aspergillus often helpful in the diagnosis of aspergillosis in non immunocompromised patients
bull Immunodiffusion indirect haemagglutination ELISA
bull The role is still uncertain
bull Positive ID test is not a proof of infection negative result can not exclude the diagnosis
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
SEROLOGICAL TESTS
bull Antigen detection (galactomannan) in blood and other body fluids rapid diagnosis of aspergilllosis in immunocompromised patients
bull Galactomannan fluctuate during the course of the infection useful if performed in a regular basis (twice per week)
bull Negative not exclude the diagnosis
bull ELISA for the detection of galactomannan has been approved for diagnostic use (the accuracy was moderate)
Christopher D Pfeiffer et al 2006
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
MOLECULAR DIAGNOSTICS
bull PCR for detection of Aspergillus in blood BAL serum etc
bull Appear promising for rapid detection of Aspergillus directly on tissue or body fluids but
ndash False positive
ndash No standardized commercial method
bull Routine use of PCR can not be recommended
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
Mark Reinwald et al
bull ldquoTreatment with mould-active antifungals prior to BAL sampling significantly decreases the performance of the Aspergillus PCR assay in haematological patients if BAL was performed after administration of more than one antifungal agentrdquo
bull ldquoThe effect of antifungal therapy on other surrogate parameters suggests reduced sensitivity rates with antifungal therapy for GM whereas no influence of antifungal therapy on β-D-glucan performance in a (single-centre retrospective) study was describedrdquo
J Antimicrob Chemother 2012 67 2260ndash2267
CONCLUSION
bull Various detection methods of agents responsible for invasive fungal infections have been developed
bull The lack of sensitivity and specificity of the test uncertainty in diagnosing IFI decrease successful treatment
bull Need further studies to improve and develop laboratory diagnostic tool for IFI
CULTURE
bull Isolation of Candida sp from sputum and faecal specimens can not be considered diagnostic of infection
bull Deep-seated infection Candida sp isolated from blood or normally sterile body fluids tissuesaspirates from other closed sites
bull Specimens should be examined as soon as possible after collection to avoid missinterpretation due to other organisms multiplication
bull Different susceptibility among Candida sp completed identification (speciation) before treatment
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
CULTURE
bull Blood cultures are positive in no more than 50 of neutropenic patients with dissemintaed candidosis or 80 of patients with endocarditis
bull Isolation of Candida sp from urine may indicate a serious infection
bull Isolation of Candida sp from LCS evidence for diagnosis of meningitis
bull Candida sp isolated from BAL in a patient with pulmonary infiltrates not sufficient for diagnosing candidosis
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
SEROLOGICAL TESTS
bull Limited usefulness
bull False positive (mucosal colonization superficial infection)
bull False negative (immunocompromised)
bull Antigen detection mannan insensitive
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
DETECTION OF D-ARABINITOL
bull D-arabinitol is produced by most of Candida sp (except C krusei and may be C glabrata)
bull It can be detected in serum or urine
bull Candidemia increased of serum D-arabiniitol-creatinine ratios has still to achieve widespread clinical use
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
MOLECULAR ASSAY
bull PCR detection of Candida DNA in blood serum LCS other specimens
bull Promising tools for rapid diagnosis of Candida sp from tissue or body-fluid
bull False positive
bull Standardized commercial assays were not available
bull Can not be recommended in diagnosis of invasive candidosis
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
The sources of yeast isolated
bull Sputum 49
bull Urine 13
bull Vaginal swab 9
bull Blood 8
bull Bronchial lavage 7
2010
bull Sputum 62
bull Vaginal swab 16
bull Blood 14
bull Urine 9
bull Bronchial lavage 5
2011
bull Sputum 102
bull Urine 18
bull Blood16 bull Bronchial lavage 12 bull Vaginal swab 11
2012
Candida spp isolated from blood
2010 2011 2012
C albicans 1 1 3
C parapsilosis 3 4 1
C pelliculosis 1 0 0
C lusitaniae 1 0 0
C tropicalis 0 2 10
C glabrata 0 1 0
C gulliermondii 0 1 1
Candida spp 1 0 0
7 9 15
Resistance pattern of Candida spp isolated from blood
2010 2011 2012
FCA ITR VRC FCA ITR VRC FCA ITR VRC
C albicans 17 115 115 115
C glabrata 19
C tropicalis 215 315 115
INVASIVE ASPERGILLOSIS
bull Incidence varies in ICU 58
bull In ICU risk factors including chronic obstructive pulmonary diseases and liver failure
bull Positive respiratory culture uncertain
bull Low sensitivity and specificity
bull Detection of galactomannan in BAL promising results
LABORATORY DIAGNOSTIC
bull Microscopy
bull Culture
bull Serological tests
bull Molecular diagnostics
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
MICROSCOPY
bull Histopathological examination of tissue biopsies immunohistochemical staining procedure
bull Non-pigmented septate hyphae dichotomous branching
bull Direct examination of sputum may help in diagnosing invasive aspergillosis BAL is better
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
A fumigatus
Richard C Barton Hindawi Publishing Corporation Scienti1048853 ca Volume 2013 Article ID 459405 29 pages httpdxdoiorg1011552013459405
CULTURE
bull Not sensitive interpreted with caution (contamination)
bull Recovered from blood LCS
bull Aspergillus isolated from BALsputum obtained from patients with a pulmonary infiltrates indicative of infection
bull A fumigatus can be isolated from sputum of patients suffer from allergic bronchopulmonary aspergillosis
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
SEROLOGICAL TESTS
bull Antobodies againts Aspergillus often helpful in the diagnosis of aspergillosis in non immunocompromised patients
bull Immunodiffusion indirect haemagglutination ELISA
bull The role is still uncertain
bull Positive ID test is not a proof of infection negative result can not exclude the diagnosis
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
SEROLOGICAL TESTS
bull Antigen detection (galactomannan) in blood and other body fluids rapid diagnosis of aspergilllosis in immunocompromised patients
bull Galactomannan fluctuate during the course of the infection useful if performed in a regular basis (twice per week)
bull Negative not exclude the diagnosis
bull ELISA for the detection of galactomannan has been approved for diagnostic use (the accuracy was moderate)
Christopher D Pfeiffer et al 2006
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
MOLECULAR DIAGNOSTICS
bull PCR for detection of Aspergillus in blood BAL serum etc
bull Appear promising for rapid detection of Aspergillus directly on tissue or body fluids but
ndash False positive
ndash No standardized commercial method
bull Routine use of PCR can not be recommended
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
Mark Reinwald et al
bull ldquoTreatment with mould-active antifungals prior to BAL sampling significantly decreases the performance of the Aspergillus PCR assay in haematological patients if BAL was performed after administration of more than one antifungal agentrdquo
bull ldquoThe effect of antifungal therapy on other surrogate parameters suggests reduced sensitivity rates with antifungal therapy for GM whereas no influence of antifungal therapy on β-D-glucan performance in a (single-centre retrospective) study was describedrdquo
J Antimicrob Chemother 2012 67 2260ndash2267
CONCLUSION
bull Various detection methods of agents responsible for invasive fungal infections have been developed
bull The lack of sensitivity and specificity of the test uncertainty in diagnosing IFI decrease successful treatment
bull Need further studies to improve and develop laboratory diagnostic tool for IFI
CULTURE
bull Blood cultures are positive in no more than 50 of neutropenic patients with dissemintaed candidosis or 80 of patients with endocarditis
bull Isolation of Candida sp from urine may indicate a serious infection
bull Isolation of Candida sp from LCS evidence for diagnosis of meningitis
bull Candida sp isolated from BAL in a patient with pulmonary infiltrates not sufficient for diagnosing candidosis
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
SEROLOGICAL TESTS
bull Limited usefulness
bull False positive (mucosal colonization superficial infection)
bull False negative (immunocompromised)
bull Antigen detection mannan insensitive
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
DETECTION OF D-ARABINITOL
bull D-arabinitol is produced by most of Candida sp (except C krusei and may be C glabrata)
bull It can be detected in serum or urine
bull Candidemia increased of serum D-arabiniitol-creatinine ratios has still to achieve widespread clinical use
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
MOLECULAR ASSAY
bull PCR detection of Candida DNA in blood serum LCS other specimens
bull Promising tools for rapid diagnosis of Candida sp from tissue or body-fluid
bull False positive
bull Standardized commercial assays were not available
bull Can not be recommended in diagnosis of invasive candidosis
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
The sources of yeast isolated
bull Sputum 49
bull Urine 13
bull Vaginal swab 9
bull Blood 8
bull Bronchial lavage 7
2010
bull Sputum 62
bull Vaginal swab 16
bull Blood 14
bull Urine 9
bull Bronchial lavage 5
2011
bull Sputum 102
bull Urine 18
bull Blood16 bull Bronchial lavage 12 bull Vaginal swab 11
2012
Candida spp isolated from blood
2010 2011 2012
C albicans 1 1 3
C parapsilosis 3 4 1
C pelliculosis 1 0 0
C lusitaniae 1 0 0
C tropicalis 0 2 10
C glabrata 0 1 0
C gulliermondii 0 1 1
Candida spp 1 0 0
7 9 15
Resistance pattern of Candida spp isolated from blood
2010 2011 2012
FCA ITR VRC FCA ITR VRC FCA ITR VRC
C albicans 17 115 115 115
C glabrata 19
C tropicalis 215 315 115
INVASIVE ASPERGILLOSIS
bull Incidence varies in ICU 58
bull In ICU risk factors including chronic obstructive pulmonary diseases and liver failure
bull Positive respiratory culture uncertain
bull Low sensitivity and specificity
bull Detection of galactomannan in BAL promising results
LABORATORY DIAGNOSTIC
bull Microscopy
bull Culture
bull Serological tests
bull Molecular diagnostics
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
MICROSCOPY
bull Histopathological examination of tissue biopsies immunohistochemical staining procedure
bull Non-pigmented septate hyphae dichotomous branching
bull Direct examination of sputum may help in diagnosing invasive aspergillosis BAL is better
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
A fumigatus
Richard C Barton Hindawi Publishing Corporation Scienti1048853 ca Volume 2013 Article ID 459405 29 pages httpdxdoiorg1011552013459405
CULTURE
bull Not sensitive interpreted with caution (contamination)
bull Recovered from blood LCS
bull Aspergillus isolated from BALsputum obtained from patients with a pulmonary infiltrates indicative of infection
bull A fumigatus can be isolated from sputum of patients suffer from allergic bronchopulmonary aspergillosis
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
SEROLOGICAL TESTS
bull Antobodies againts Aspergillus often helpful in the diagnosis of aspergillosis in non immunocompromised patients
bull Immunodiffusion indirect haemagglutination ELISA
bull The role is still uncertain
bull Positive ID test is not a proof of infection negative result can not exclude the diagnosis
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
SEROLOGICAL TESTS
bull Antigen detection (galactomannan) in blood and other body fluids rapid diagnosis of aspergilllosis in immunocompromised patients
bull Galactomannan fluctuate during the course of the infection useful if performed in a regular basis (twice per week)
bull Negative not exclude the diagnosis
bull ELISA for the detection of galactomannan has been approved for diagnostic use (the accuracy was moderate)
Christopher D Pfeiffer et al 2006
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
MOLECULAR DIAGNOSTICS
bull PCR for detection of Aspergillus in blood BAL serum etc
bull Appear promising for rapid detection of Aspergillus directly on tissue or body fluids but
ndash False positive
ndash No standardized commercial method
bull Routine use of PCR can not be recommended
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
Mark Reinwald et al
bull ldquoTreatment with mould-active antifungals prior to BAL sampling significantly decreases the performance of the Aspergillus PCR assay in haematological patients if BAL was performed after administration of more than one antifungal agentrdquo
bull ldquoThe effect of antifungal therapy on other surrogate parameters suggests reduced sensitivity rates with antifungal therapy for GM whereas no influence of antifungal therapy on β-D-glucan performance in a (single-centre retrospective) study was describedrdquo
J Antimicrob Chemother 2012 67 2260ndash2267
CONCLUSION
bull Various detection methods of agents responsible for invasive fungal infections have been developed
bull The lack of sensitivity and specificity of the test uncertainty in diagnosing IFI decrease successful treatment
bull Need further studies to improve and develop laboratory diagnostic tool for IFI
SEROLOGICAL TESTS
bull Limited usefulness
bull False positive (mucosal colonization superficial infection)
bull False negative (immunocompromised)
bull Antigen detection mannan insensitive
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
DETECTION OF D-ARABINITOL
bull D-arabinitol is produced by most of Candida sp (except C krusei and may be C glabrata)
bull It can be detected in serum or urine
bull Candidemia increased of serum D-arabiniitol-creatinine ratios has still to achieve widespread clinical use
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
MOLECULAR ASSAY
bull PCR detection of Candida DNA in blood serum LCS other specimens
bull Promising tools for rapid diagnosis of Candida sp from tissue or body-fluid
bull False positive
bull Standardized commercial assays were not available
bull Can not be recommended in diagnosis of invasive candidosis
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
The sources of yeast isolated
bull Sputum 49
bull Urine 13
bull Vaginal swab 9
bull Blood 8
bull Bronchial lavage 7
2010
bull Sputum 62
bull Vaginal swab 16
bull Blood 14
bull Urine 9
bull Bronchial lavage 5
2011
bull Sputum 102
bull Urine 18
bull Blood16 bull Bronchial lavage 12 bull Vaginal swab 11
2012
Candida spp isolated from blood
2010 2011 2012
C albicans 1 1 3
C parapsilosis 3 4 1
C pelliculosis 1 0 0
C lusitaniae 1 0 0
C tropicalis 0 2 10
C glabrata 0 1 0
C gulliermondii 0 1 1
Candida spp 1 0 0
7 9 15
Resistance pattern of Candida spp isolated from blood
2010 2011 2012
FCA ITR VRC FCA ITR VRC FCA ITR VRC
C albicans 17 115 115 115
C glabrata 19
C tropicalis 215 315 115
INVASIVE ASPERGILLOSIS
bull Incidence varies in ICU 58
bull In ICU risk factors including chronic obstructive pulmonary diseases and liver failure
bull Positive respiratory culture uncertain
bull Low sensitivity and specificity
bull Detection of galactomannan in BAL promising results
LABORATORY DIAGNOSTIC
bull Microscopy
bull Culture
bull Serological tests
bull Molecular diagnostics
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
MICROSCOPY
bull Histopathological examination of tissue biopsies immunohistochemical staining procedure
bull Non-pigmented septate hyphae dichotomous branching
bull Direct examination of sputum may help in diagnosing invasive aspergillosis BAL is better
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
A fumigatus
Richard C Barton Hindawi Publishing Corporation Scienti1048853 ca Volume 2013 Article ID 459405 29 pages httpdxdoiorg1011552013459405
CULTURE
bull Not sensitive interpreted with caution (contamination)
bull Recovered from blood LCS
bull Aspergillus isolated from BALsputum obtained from patients with a pulmonary infiltrates indicative of infection
bull A fumigatus can be isolated from sputum of patients suffer from allergic bronchopulmonary aspergillosis
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
SEROLOGICAL TESTS
bull Antobodies againts Aspergillus often helpful in the diagnosis of aspergillosis in non immunocompromised patients
bull Immunodiffusion indirect haemagglutination ELISA
bull The role is still uncertain
bull Positive ID test is not a proof of infection negative result can not exclude the diagnosis
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
SEROLOGICAL TESTS
bull Antigen detection (galactomannan) in blood and other body fluids rapid diagnosis of aspergilllosis in immunocompromised patients
bull Galactomannan fluctuate during the course of the infection useful if performed in a regular basis (twice per week)
bull Negative not exclude the diagnosis
bull ELISA for the detection of galactomannan has been approved for diagnostic use (the accuracy was moderate)
Christopher D Pfeiffer et al 2006
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
MOLECULAR DIAGNOSTICS
bull PCR for detection of Aspergillus in blood BAL serum etc
bull Appear promising for rapid detection of Aspergillus directly on tissue or body fluids but
ndash False positive
ndash No standardized commercial method
bull Routine use of PCR can not be recommended
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
Mark Reinwald et al
bull ldquoTreatment with mould-active antifungals prior to BAL sampling significantly decreases the performance of the Aspergillus PCR assay in haematological patients if BAL was performed after administration of more than one antifungal agentrdquo
bull ldquoThe effect of antifungal therapy on other surrogate parameters suggests reduced sensitivity rates with antifungal therapy for GM whereas no influence of antifungal therapy on β-D-glucan performance in a (single-centre retrospective) study was describedrdquo
J Antimicrob Chemother 2012 67 2260ndash2267
CONCLUSION
bull Various detection methods of agents responsible for invasive fungal infections have been developed
bull The lack of sensitivity and specificity of the test uncertainty in diagnosing IFI decrease successful treatment
bull Need further studies to improve and develop laboratory diagnostic tool for IFI
DETECTION OF D-ARABINITOL
bull D-arabinitol is produced by most of Candida sp (except C krusei and may be C glabrata)
bull It can be detected in serum or urine
bull Candidemia increased of serum D-arabiniitol-creatinine ratios has still to achieve widespread clinical use
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
MOLECULAR ASSAY
bull PCR detection of Candida DNA in blood serum LCS other specimens
bull Promising tools for rapid diagnosis of Candida sp from tissue or body-fluid
bull False positive
bull Standardized commercial assays were not available
bull Can not be recommended in diagnosis of invasive candidosis
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
The sources of yeast isolated
bull Sputum 49
bull Urine 13
bull Vaginal swab 9
bull Blood 8
bull Bronchial lavage 7
2010
bull Sputum 62
bull Vaginal swab 16
bull Blood 14
bull Urine 9
bull Bronchial lavage 5
2011
bull Sputum 102
bull Urine 18
bull Blood16 bull Bronchial lavage 12 bull Vaginal swab 11
2012
Candida spp isolated from blood
2010 2011 2012
C albicans 1 1 3
C parapsilosis 3 4 1
C pelliculosis 1 0 0
C lusitaniae 1 0 0
C tropicalis 0 2 10
C glabrata 0 1 0
C gulliermondii 0 1 1
Candida spp 1 0 0
7 9 15
Resistance pattern of Candida spp isolated from blood
2010 2011 2012
FCA ITR VRC FCA ITR VRC FCA ITR VRC
C albicans 17 115 115 115
C glabrata 19
C tropicalis 215 315 115
INVASIVE ASPERGILLOSIS
bull Incidence varies in ICU 58
bull In ICU risk factors including chronic obstructive pulmonary diseases and liver failure
bull Positive respiratory culture uncertain
bull Low sensitivity and specificity
bull Detection of galactomannan in BAL promising results
LABORATORY DIAGNOSTIC
bull Microscopy
bull Culture
bull Serological tests
bull Molecular diagnostics
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
MICROSCOPY
bull Histopathological examination of tissue biopsies immunohistochemical staining procedure
bull Non-pigmented septate hyphae dichotomous branching
bull Direct examination of sputum may help in diagnosing invasive aspergillosis BAL is better
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
A fumigatus
Richard C Barton Hindawi Publishing Corporation Scienti1048853 ca Volume 2013 Article ID 459405 29 pages httpdxdoiorg1011552013459405
CULTURE
bull Not sensitive interpreted with caution (contamination)
bull Recovered from blood LCS
bull Aspergillus isolated from BALsputum obtained from patients with a pulmonary infiltrates indicative of infection
bull A fumigatus can be isolated from sputum of patients suffer from allergic bronchopulmonary aspergillosis
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
SEROLOGICAL TESTS
bull Antobodies againts Aspergillus often helpful in the diagnosis of aspergillosis in non immunocompromised patients
bull Immunodiffusion indirect haemagglutination ELISA
bull The role is still uncertain
bull Positive ID test is not a proof of infection negative result can not exclude the diagnosis
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
SEROLOGICAL TESTS
bull Antigen detection (galactomannan) in blood and other body fluids rapid diagnosis of aspergilllosis in immunocompromised patients
bull Galactomannan fluctuate during the course of the infection useful if performed in a regular basis (twice per week)
bull Negative not exclude the diagnosis
bull ELISA for the detection of galactomannan has been approved for diagnostic use (the accuracy was moderate)
Christopher D Pfeiffer et al 2006
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
MOLECULAR DIAGNOSTICS
bull PCR for detection of Aspergillus in blood BAL serum etc
bull Appear promising for rapid detection of Aspergillus directly on tissue or body fluids but
ndash False positive
ndash No standardized commercial method
bull Routine use of PCR can not be recommended
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
Mark Reinwald et al
bull ldquoTreatment with mould-active antifungals prior to BAL sampling significantly decreases the performance of the Aspergillus PCR assay in haematological patients if BAL was performed after administration of more than one antifungal agentrdquo
bull ldquoThe effect of antifungal therapy on other surrogate parameters suggests reduced sensitivity rates with antifungal therapy for GM whereas no influence of antifungal therapy on β-D-glucan performance in a (single-centre retrospective) study was describedrdquo
J Antimicrob Chemother 2012 67 2260ndash2267
CONCLUSION
bull Various detection methods of agents responsible for invasive fungal infections have been developed
bull The lack of sensitivity and specificity of the test uncertainty in diagnosing IFI decrease successful treatment
bull Need further studies to improve and develop laboratory diagnostic tool for IFI
MOLECULAR ASSAY
bull PCR detection of Candida DNA in blood serum LCS other specimens
bull Promising tools for rapid diagnosis of Candida sp from tissue or body-fluid
bull False positive
bull Standardized commercial assays were not available
bull Can not be recommended in diagnosis of invasive candidosis
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
The sources of yeast isolated
bull Sputum 49
bull Urine 13
bull Vaginal swab 9
bull Blood 8
bull Bronchial lavage 7
2010
bull Sputum 62
bull Vaginal swab 16
bull Blood 14
bull Urine 9
bull Bronchial lavage 5
2011
bull Sputum 102
bull Urine 18
bull Blood16 bull Bronchial lavage 12 bull Vaginal swab 11
2012
Candida spp isolated from blood
2010 2011 2012
C albicans 1 1 3
C parapsilosis 3 4 1
C pelliculosis 1 0 0
C lusitaniae 1 0 0
C tropicalis 0 2 10
C glabrata 0 1 0
C gulliermondii 0 1 1
Candida spp 1 0 0
7 9 15
Resistance pattern of Candida spp isolated from blood
2010 2011 2012
FCA ITR VRC FCA ITR VRC FCA ITR VRC
C albicans 17 115 115 115
C glabrata 19
C tropicalis 215 315 115
INVASIVE ASPERGILLOSIS
bull Incidence varies in ICU 58
bull In ICU risk factors including chronic obstructive pulmonary diseases and liver failure
bull Positive respiratory culture uncertain
bull Low sensitivity and specificity
bull Detection of galactomannan in BAL promising results
LABORATORY DIAGNOSTIC
bull Microscopy
bull Culture
bull Serological tests
bull Molecular diagnostics
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
MICROSCOPY
bull Histopathological examination of tissue biopsies immunohistochemical staining procedure
bull Non-pigmented septate hyphae dichotomous branching
bull Direct examination of sputum may help in diagnosing invasive aspergillosis BAL is better
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
A fumigatus
Richard C Barton Hindawi Publishing Corporation Scienti1048853 ca Volume 2013 Article ID 459405 29 pages httpdxdoiorg1011552013459405
CULTURE
bull Not sensitive interpreted with caution (contamination)
bull Recovered from blood LCS
bull Aspergillus isolated from BALsputum obtained from patients with a pulmonary infiltrates indicative of infection
bull A fumigatus can be isolated from sputum of patients suffer from allergic bronchopulmonary aspergillosis
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
SEROLOGICAL TESTS
bull Antobodies againts Aspergillus often helpful in the diagnosis of aspergillosis in non immunocompromised patients
bull Immunodiffusion indirect haemagglutination ELISA
bull The role is still uncertain
bull Positive ID test is not a proof of infection negative result can not exclude the diagnosis
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
SEROLOGICAL TESTS
bull Antigen detection (galactomannan) in blood and other body fluids rapid diagnosis of aspergilllosis in immunocompromised patients
bull Galactomannan fluctuate during the course of the infection useful if performed in a regular basis (twice per week)
bull Negative not exclude the diagnosis
bull ELISA for the detection of galactomannan has been approved for diagnostic use (the accuracy was moderate)
Christopher D Pfeiffer et al 2006
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
MOLECULAR DIAGNOSTICS
bull PCR for detection of Aspergillus in blood BAL serum etc
bull Appear promising for rapid detection of Aspergillus directly on tissue or body fluids but
ndash False positive
ndash No standardized commercial method
bull Routine use of PCR can not be recommended
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
Mark Reinwald et al
bull ldquoTreatment with mould-active antifungals prior to BAL sampling significantly decreases the performance of the Aspergillus PCR assay in haematological patients if BAL was performed after administration of more than one antifungal agentrdquo
bull ldquoThe effect of antifungal therapy on other surrogate parameters suggests reduced sensitivity rates with antifungal therapy for GM whereas no influence of antifungal therapy on β-D-glucan performance in a (single-centre retrospective) study was describedrdquo
J Antimicrob Chemother 2012 67 2260ndash2267
CONCLUSION
bull Various detection methods of agents responsible for invasive fungal infections have been developed
bull The lack of sensitivity and specificity of the test uncertainty in diagnosing IFI decrease successful treatment
bull Need further studies to improve and develop laboratory diagnostic tool for IFI
The sources of yeast isolated
bull Sputum 49
bull Urine 13
bull Vaginal swab 9
bull Blood 8
bull Bronchial lavage 7
2010
bull Sputum 62
bull Vaginal swab 16
bull Blood 14
bull Urine 9
bull Bronchial lavage 5
2011
bull Sputum 102
bull Urine 18
bull Blood16 bull Bronchial lavage 12 bull Vaginal swab 11
2012
Candida spp isolated from blood
2010 2011 2012
C albicans 1 1 3
C parapsilosis 3 4 1
C pelliculosis 1 0 0
C lusitaniae 1 0 0
C tropicalis 0 2 10
C glabrata 0 1 0
C gulliermondii 0 1 1
Candida spp 1 0 0
7 9 15
Resistance pattern of Candida spp isolated from blood
2010 2011 2012
FCA ITR VRC FCA ITR VRC FCA ITR VRC
C albicans 17 115 115 115
C glabrata 19
C tropicalis 215 315 115
INVASIVE ASPERGILLOSIS
bull Incidence varies in ICU 58
bull In ICU risk factors including chronic obstructive pulmonary diseases and liver failure
bull Positive respiratory culture uncertain
bull Low sensitivity and specificity
bull Detection of galactomannan in BAL promising results
LABORATORY DIAGNOSTIC
bull Microscopy
bull Culture
bull Serological tests
bull Molecular diagnostics
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
MICROSCOPY
bull Histopathological examination of tissue biopsies immunohistochemical staining procedure
bull Non-pigmented septate hyphae dichotomous branching
bull Direct examination of sputum may help in diagnosing invasive aspergillosis BAL is better
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
A fumigatus
Richard C Barton Hindawi Publishing Corporation Scienti1048853 ca Volume 2013 Article ID 459405 29 pages httpdxdoiorg1011552013459405
CULTURE
bull Not sensitive interpreted with caution (contamination)
bull Recovered from blood LCS
bull Aspergillus isolated from BALsputum obtained from patients with a pulmonary infiltrates indicative of infection
bull A fumigatus can be isolated from sputum of patients suffer from allergic bronchopulmonary aspergillosis
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
SEROLOGICAL TESTS
bull Antobodies againts Aspergillus often helpful in the diagnosis of aspergillosis in non immunocompromised patients
bull Immunodiffusion indirect haemagglutination ELISA
bull The role is still uncertain
bull Positive ID test is not a proof of infection negative result can not exclude the diagnosis
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
SEROLOGICAL TESTS
bull Antigen detection (galactomannan) in blood and other body fluids rapid diagnosis of aspergilllosis in immunocompromised patients
bull Galactomannan fluctuate during the course of the infection useful if performed in a regular basis (twice per week)
bull Negative not exclude the diagnosis
bull ELISA for the detection of galactomannan has been approved for diagnostic use (the accuracy was moderate)
Christopher D Pfeiffer et al 2006
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
MOLECULAR DIAGNOSTICS
bull PCR for detection of Aspergillus in blood BAL serum etc
bull Appear promising for rapid detection of Aspergillus directly on tissue or body fluids but
ndash False positive
ndash No standardized commercial method
bull Routine use of PCR can not be recommended
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
Mark Reinwald et al
bull ldquoTreatment with mould-active antifungals prior to BAL sampling significantly decreases the performance of the Aspergillus PCR assay in haematological patients if BAL was performed after administration of more than one antifungal agentrdquo
bull ldquoThe effect of antifungal therapy on other surrogate parameters suggests reduced sensitivity rates with antifungal therapy for GM whereas no influence of antifungal therapy on β-D-glucan performance in a (single-centre retrospective) study was describedrdquo
J Antimicrob Chemother 2012 67 2260ndash2267
CONCLUSION
bull Various detection methods of agents responsible for invasive fungal infections have been developed
bull The lack of sensitivity and specificity of the test uncertainty in diagnosing IFI decrease successful treatment
bull Need further studies to improve and develop laboratory diagnostic tool for IFI
Candida spp isolated from blood
2010 2011 2012
C albicans 1 1 3
C parapsilosis 3 4 1
C pelliculosis 1 0 0
C lusitaniae 1 0 0
C tropicalis 0 2 10
C glabrata 0 1 0
C gulliermondii 0 1 1
Candida spp 1 0 0
7 9 15
Resistance pattern of Candida spp isolated from blood
2010 2011 2012
FCA ITR VRC FCA ITR VRC FCA ITR VRC
C albicans 17 115 115 115
C glabrata 19
C tropicalis 215 315 115
INVASIVE ASPERGILLOSIS
bull Incidence varies in ICU 58
bull In ICU risk factors including chronic obstructive pulmonary diseases and liver failure
bull Positive respiratory culture uncertain
bull Low sensitivity and specificity
bull Detection of galactomannan in BAL promising results
LABORATORY DIAGNOSTIC
bull Microscopy
bull Culture
bull Serological tests
bull Molecular diagnostics
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
MICROSCOPY
bull Histopathological examination of tissue biopsies immunohistochemical staining procedure
bull Non-pigmented septate hyphae dichotomous branching
bull Direct examination of sputum may help in diagnosing invasive aspergillosis BAL is better
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
A fumigatus
Richard C Barton Hindawi Publishing Corporation Scienti1048853 ca Volume 2013 Article ID 459405 29 pages httpdxdoiorg1011552013459405
CULTURE
bull Not sensitive interpreted with caution (contamination)
bull Recovered from blood LCS
bull Aspergillus isolated from BALsputum obtained from patients with a pulmonary infiltrates indicative of infection
bull A fumigatus can be isolated from sputum of patients suffer from allergic bronchopulmonary aspergillosis
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
SEROLOGICAL TESTS
bull Antobodies againts Aspergillus often helpful in the diagnosis of aspergillosis in non immunocompromised patients
bull Immunodiffusion indirect haemagglutination ELISA
bull The role is still uncertain
bull Positive ID test is not a proof of infection negative result can not exclude the diagnosis
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
SEROLOGICAL TESTS
bull Antigen detection (galactomannan) in blood and other body fluids rapid diagnosis of aspergilllosis in immunocompromised patients
bull Galactomannan fluctuate during the course of the infection useful if performed in a regular basis (twice per week)
bull Negative not exclude the diagnosis
bull ELISA for the detection of galactomannan has been approved for diagnostic use (the accuracy was moderate)
Christopher D Pfeiffer et al 2006
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
MOLECULAR DIAGNOSTICS
bull PCR for detection of Aspergillus in blood BAL serum etc
bull Appear promising for rapid detection of Aspergillus directly on tissue or body fluids but
ndash False positive
ndash No standardized commercial method
bull Routine use of PCR can not be recommended
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
Mark Reinwald et al
bull ldquoTreatment with mould-active antifungals prior to BAL sampling significantly decreases the performance of the Aspergillus PCR assay in haematological patients if BAL was performed after administration of more than one antifungal agentrdquo
bull ldquoThe effect of antifungal therapy on other surrogate parameters suggests reduced sensitivity rates with antifungal therapy for GM whereas no influence of antifungal therapy on β-D-glucan performance in a (single-centre retrospective) study was describedrdquo
J Antimicrob Chemother 2012 67 2260ndash2267
CONCLUSION
bull Various detection methods of agents responsible for invasive fungal infections have been developed
bull The lack of sensitivity and specificity of the test uncertainty in diagnosing IFI decrease successful treatment
bull Need further studies to improve and develop laboratory diagnostic tool for IFI
Resistance pattern of Candida spp isolated from blood
2010 2011 2012
FCA ITR VRC FCA ITR VRC FCA ITR VRC
C albicans 17 115 115 115
C glabrata 19
C tropicalis 215 315 115
INVASIVE ASPERGILLOSIS
bull Incidence varies in ICU 58
bull In ICU risk factors including chronic obstructive pulmonary diseases and liver failure
bull Positive respiratory culture uncertain
bull Low sensitivity and specificity
bull Detection of galactomannan in BAL promising results
LABORATORY DIAGNOSTIC
bull Microscopy
bull Culture
bull Serological tests
bull Molecular diagnostics
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
MICROSCOPY
bull Histopathological examination of tissue biopsies immunohistochemical staining procedure
bull Non-pigmented septate hyphae dichotomous branching
bull Direct examination of sputum may help in diagnosing invasive aspergillosis BAL is better
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
A fumigatus
Richard C Barton Hindawi Publishing Corporation Scienti1048853 ca Volume 2013 Article ID 459405 29 pages httpdxdoiorg1011552013459405
CULTURE
bull Not sensitive interpreted with caution (contamination)
bull Recovered from blood LCS
bull Aspergillus isolated from BALsputum obtained from patients with a pulmonary infiltrates indicative of infection
bull A fumigatus can be isolated from sputum of patients suffer from allergic bronchopulmonary aspergillosis
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
SEROLOGICAL TESTS
bull Antobodies againts Aspergillus often helpful in the diagnosis of aspergillosis in non immunocompromised patients
bull Immunodiffusion indirect haemagglutination ELISA
bull The role is still uncertain
bull Positive ID test is not a proof of infection negative result can not exclude the diagnosis
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
SEROLOGICAL TESTS
bull Antigen detection (galactomannan) in blood and other body fluids rapid diagnosis of aspergilllosis in immunocompromised patients
bull Galactomannan fluctuate during the course of the infection useful if performed in a regular basis (twice per week)
bull Negative not exclude the diagnosis
bull ELISA for the detection of galactomannan has been approved for diagnostic use (the accuracy was moderate)
Christopher D Pfeiffer et al 2006
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
MOLECULAR DIAGNOSTICS
bull PCR for detection of Aspergillus in blood BAL serum etc
bull Appear promising for rapid detection of Aspergillus directly on tissue or body fluids but
ndash False positive
ndash No standardized commercial method
bull Routine use of PCR can not be recommended
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
Mark Reinwald et al
bull ldquoTreatment with mould-active antifungals prior to BAL sampling significantly decreases the performance of the Aspergillus PCR assay in haematological patients if BAL was performed after administration of more than one antifungal agentrdquo
bull ldquoThe effect of antifungal therapy on other surrogate parameters suggests reduced sensitivity rates with antifungal therapy for GM whereas no influence of antifungal therapy on β-D-glucan performance in a (single-centre retrospective) study was describedrdquo
J Antimicrob Chemother 2012 67 2260ndash2267
CONCLUSION
bull Various detection methods of agents responsible for invasive fungal infections have been developed
bull The lack of sensitivity and specificity of the test uncertainty in diagnosing IFI decrease successful treatment
bull Need further studies to improve and develop laboratory diagnostic tool for IFI
INVASIVE ASPERGILLOSIS
bull Incidence varies in ICU 58
bull In ICU risk factors including chronic obstructive pulmonary diseases and liver failure
bull Positive respiratory culture uncertain
bull Low sensitivity and specificity
bull Detection of galactomannan in BAL promising results
LABORATORY DIAGNOSTIC
bull Microscopy
bull Culture
bull Serological tests
bull Molecular diagnostics
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
MICROSCOPY
bull Histopathological examination of tissue biopsies immunohistochemical staining procedure
bull Non-pigmented septate hyphae dichotomous branching
bull Direct examination of sputum may help in diagnosing invasive aspergillosis BAL is better
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
A fumigatus
Richard C Barton Hindawi Publishing Corporation Scienti1048853 ca Volume 2013 Article ID 459405 29 pages httpdxdoiorg1011552013459405
CULTURE
bull Not sensitive interpreted with caution (contamination)
bull Recovered from blood LCS
bull Aspergillus isolated from BALsputum obtained from patients with a pulmonary infiltrates indicative of infection
bull A fumigatus can be isolated from sputum of patients suffer from allergic bronchopulmonary aspergillosis
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
SEROLOGICAL TESTS
bull Antobodies againts Aspergillus often helpful in the diagnosis of aspergillosis in non immunocompromised patients
bull Immunodiffusion indirect haemagglutination ELISA
bull The role is still uncertain
bull Positive ID test is not a proof of infection negative result can not exclude the diagnosis
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
SEROLOGICAL TESTS
bull Antigen detection (galactomannan) in blood and other body fluids rapid diagnosis of aspergilllosis in immunocompromised patients
bull Galactomannan fluctuate during the course of the infection useful if performed in a regular basis (twice per week)
bull Negative not exclude the diagnosis
bull ELISA for the detection of galactomannan has been approved for diagnostic use (the accuracy was moderate)
Christopher D Pfeiffer et al 2006
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
MOLECULAR DIAGNOSTICS
bull PCR for detection of Aspergillus in blood BAL serum etc
bull Appear promising for rapid detection of Aspergillus directly on tissue or body fluids but
ndash False positive
ndash No standardized commercial method
bull Routine use of PCR can not be recommended
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
Mark Reinwald et al
bull ldquoTreatment with mould-active antifungals prior to BAL sampling significantly decreases the performance of the Aspergillus PCR assay in haematological patients if BAL was performed after administration of more than one antifungal agentrdquo
bull ldquoThe effect of antifungal therapy on other surrogate parameters suggests reduced sensitivity rates with antifungal therapy for GM whereas no influence of antifungal therapy on β-D-glucan performance in a (single-centre retrospective) study was describedrdquo
J Antimicrob Chemother 2012 67 2260ndash2267
CONCLUSION
bull Various detection methods of agents responsible for invasive fungal infections have been developed
bull The lack of sensitivity and specificity of the test uncertainty in diagnosing IFI decrease successful treatment
bull Need further studies to improve and develop laboratory diagnostic tool for IFI
bull Incidence varies in ICU 58
bull In ICU risk factors including chronic obstructive pulmonary diseases and liver failure
bull Positive respiratory culture uncertain
bull Low sensitivity and specificity
bull Detection of galactomannan in BAL promising results
LABORATORY DIAGNOSTIC
bull Microscopy
bull Culture
bull Serological tests
bull Molecular diagnostics
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
MICROSCOPY
bull Histopathological examination of tissue biopsies immunohistochemical staining procedure
bull Non-pigmented septate hyphae dichotomous branching
bull Direct examination of sputum may help in diagnosing invasive aspergillosis BAL is better
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
A fumigatus
Richard C Barton Hindawi Publishing Corporation Scienti1048853 ca Volume 2013 Article ID 459405 29 pages httpdxdoiorg1011552013459405
CULTURE
bull Not sensitive interpreted with caution (contamination)
bull Recovered from blood LCS
bull Aspergillus isolated from BALsputum obtained from patients with a pulmonary infiltrates indicative of infection
bull A fumigatus can be isolated from sputum of patients suffer from allergic bronchopulmonary aspergillosis
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
SEROLOGICAL TESTS
bull Antobodies againts Aspergillus often helpful in the diagnosis of aspergillosis in non immunocompromised patients
bull Immunodiffusion indirect haemagglutination ELISA
bull The role is still uncertain
bull Positive ID test is not a proof of infection negative result can not exclude the diagnosis
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
SEROLOGICAL TESTS
bull Antigen detection (galactomannan) in blood and other body fluids rapid diagnosis of aspergilllosis in immunocompromised patients
bull Galactomannan fluctuate during the course of the infection useful if performed in a regular basis (twice per week)
bull Negative not exclude the diagnosis
bull ELISA for the detection of galactomannan has been approved for diagnostic use (the accuracy was moderate)
Christopher D Pfeiffer et al 2006
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
MOLECULAR DIAGNOSTICS
bull PCR for detection of Aspergillus in blood BAL serum etc
bull Appear promising for rapid detection of Aspergillus directly on tissue or body fluids but
ndash False positive
ndash No standardized commercial method
bull Routine use of PCR can not be recommended
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
Mark Reinwald et al
bull ldquoTreatment with mould-active antifungals prior to BAL sampling significantly decreases the performance of the Aspergillus PCR assay in haematological patients if BAL was performed after administration of more than one antifungal agentrdquo
bull ldquoThe effect of antifungal therapy on other surrogate parameters suggests reduced sensitivity rates with antifungal therapy for GM whereas no influence of antifungal therapy on β-D-glucan performance in a (single-centre retrospective) study was describedrdquo
J Antimicrob Chemother 2012 67 2260ndash2267
CONCLUSION
bull Various detection methods of agents responsible for invasive fungal infections have been developed
bull The lack of sensitivity and specificity of the test uncertainty in diagnosing IFI decrease successful treatment
bull Need further studies to improve and develop laboratory diagnostic tool for IFI
LABORATORY DIAGNOSTIC
bull Microscopy
bull Culture
bull Serological tests
bull Molecular diagnostics
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
MICROSCOPY
bull Histopathological examination of tissue biopsies immunohistochemical staining procedure
bull Non-pigmented septate hyphae dichotomous branching
bull Direct examination of sputum may help in diagnosing invasive aspergillosis BAL is better
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
A fumigatus
Richard C Barton Hindawi Publishing Corporation Scienti1048853 ca Volume 2013 Article ID 459405 29 pages httpdxdoiorg1011552013459405
CULTURE
bull Not sensitive interpreted with caution (contamination)
bull Recovered from blood LCS
bull Aspergillus isolated from BALsputum obtained from patients with a pulmonary infiltrates indicative of infection
bull A fumigatus can be isolated from sputum of patients suffer from allergic bronchopulmonary aspergillosis
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
SEROLOGICAL TESTS
bull Antobodies againts Aspergillus often helpful in the diagnosis of aspergillosis in non immunocompromised patients
bull Immunodiffusion indirect haemagglutination ELISA
bull The role is still uncertain
bull Positive ID test is not a proof of infection negative result can not exclude the diagnosis
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
SEROLOGICAL TESTS
bull Antigen detection (galactomannan) in blood and other body fluids rapid diagnosis of aspergilllosis in immunocompromised patients
bull Galactomannan fluctuate during the course of the infection useful if performed in a regular basis (twice per week)
bull Negative not exclude the diagnosis
bull ELISA for the detection of galactomannan has been approved for diagnostic use (the accuracy was moderate)
Christopher D Pfeiffer et al 2006
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
MOLECULAR DIAGNOSTICS
bull PCR for detection of Aspergillus in blood BAL serum etc
bull Appear promising for rapid detection of Aspergillus directly on tissue or body fluids but
ndash False positive
ndash No standardized commercial method
bull Routine use of PCR can not be recommended
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
Mark Reinwald et al
bull ldquoTreatment with mould-active antifungals prior to BAL sampling significantly decreases the performance of the Aspergillus PCR assay in haematological patients if BAL was performed after administration of more than one antifungal agentrdquo
bull ldquoThe effect of antifungal therapy on other surrogate parameters suggests reduced sensitivity rates with antifungal therapy for GM whereas no influence of antifungal therapy on β-D-glucan performance in a (single-centre retrospective) study was describedrdquo
J Antimicrob Chemother 2012 67 2260ndash2267
CONCLUSION
bull Various detection methods of agents responsible for invasive fungal infections have been developed
bull The lack of sensitivity and specificity of the test uncertainty in diagnosing IFI decrease successful treatment
bull Need further studies to improve and develop laboratory diagnostic tool for IFI
MICROSCOPY
bull Histopathological examination of tissue biopsies immunohistochemical staining procedure
bull Non-pigmented septate hyphae dichotomous branching
bull Direct examination of sputum may help in diagnosing invasive aspergillosis BAL is better
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
A fumigatus
Richard C Barton Hindawi Publishing Corporation Scienti1048853 ca Volume 2013 Article ID 459405 29 pages httpdxdoiorg1011552013459405
CULTURE
bull Not sensitive interpreted with caution (contamination)
bull Recovered from blood LCS
bull Aspergillus isolated from BALsputum obtained from patients with a pulmonary infiltrates indicative of infection
bull A fumigatus can be isolated from sputum of patients suffer from allergic bronchopulmonary aspergillosis
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
SEROLOGICAL TESTS
bull Antobodies againts Aspergillus often helpful in the diagnosis of aspergillosis in non immunocompromised patients
bull Immunodiffusion indirect haemagglutination ELISA
bull The role is still uncertain
bull Positive ID test is not a proof of infection negative result can not exclude the diagnosis
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
SEROLOGICAL TESTS
bull Antigen detection (galactomannan) in blood and other body fluids rapid diagnosis of aspergilllosis in immunocompromised patients
bull Galactomannan fluctuate during the course of the infection useful if performed in a regular basis (twice per week)
bull Negative not exclude the diagnosis
bull ELISA for the detection of galactomannan has been approved for diagnostic use (the accuracy was moderate)
Christopher D Pfeiffer et al 2006
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
MOLECULAR DIAGNOSTICS
bull PCR for detection of Aspergillus in blood BAL serum etc
bull Appear promising for rapid detection of Aspergillus directly on tissue or body fluids but
ndash False positive
ndash No standardized commercial method
bull Routine use of PCR can not be recommended
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
Mark Reinwald et al
bull ldquoTreatment with mould-active antifungals prior to BAL sampling significantly decreases the performance of the Aspergillus PCR assay in haematological patients if BAL was performed after administration of more than one antifungal agentrdquo
bull ldquoThe effect of antifungal therapy on other surrogate parameters suggests reduced sensitivity rates with antifungal therapy for GM whereas no influence of antifungal therapy on β-D-glucan performance in a (single-centre retrospective) study was describedrdquo
J Antimicrob Chemother 2012 67 2260ndash2267
CONCLUSION
bull Various detection methods of agents responsible for invasive fungal infections have been developed
bull The lack of sensitivity and specificity of the test uncertainty in diagnosing IFI decrease successful treatment
bull Need further studies to improve and develop laboratory diagnostic tool for IFI
A fumigatus
Richard C Barton Hindawi Publishing Corporation Scienti1048853 ca Volume 2013 Article ID 459405 29 pages httpdxdoiorg1011552013459405
CULTURE
bull Not sensitive interpreted with caution (contamination)
bull Recovered from blood LCS
bull Aspergillus isolated from BALsputum obtained from patients with a pulmonary infiltrates indicative of infection
bull A fumigatus can be isolated from sputum of patients suffer from allergic bronchopulmonary aspergillosis
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
SEROLOGICAL TESTS
bull Antobodies againts Aspergillus often helpful in the diagnosis of aspergillosis in non immunocompromised patients
bull Immunodiffusion indirect haemagglutination ELISA
bull The role is still uncertain
bull Positive ID test is not a proof of infection negative result can not exclude the diagnosis
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
SEROLOGICAL TESTS
bull Antigen detection (galactomannan) in blood and other body fluids rapid diagnosis of aspergilllosis in immunocompromised patients
bull Galactomannan fluctuate during the course of the infection useful if performed in a regular basis (twice per week)
bull Negative not exclude the diagnosis
bull ELISA for the detection of galactomannan has been approved for diagnostic use (the accuracy was moderate)
Christopher D Pfeiffer et al 2006
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
MOLECULAR DIAGNOSTICS
bull PCR for detection of Aspergillus in blood BAL serum etc
bull Appear promising for rapid detection of Aspergillus directly on tissue or body fluids but
ndash False positive
ndash No standardized commercial method
bull Routine use of PCR can not be recommended
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
Mark Reinwald et al
bull ldquoTreatment with mould-active antifungals prior to BAL sampling significantly decreases the performance of the Aspergillus PCR assay in haematological patients if BAL was performed after administration of more than one antifungal agentrdquo
bull ldquoThe effect of antifungal therapy on other surrogate parameters suggests reduced sensitivity rates with antifungal therapy for GM whereas no influence of antifungal therapy on β-D-glucan performance in a (single-centre retrospective) study was describedrdquo
J Antimicrob Chemother 2012 67 2260ndash2267
CONCLUSION
bull Various detection methods of agents responsible for invasive fungal infections have been developed
bull The lack of sensitivity and specificity of the test uncertainty in diagnosing IFI decrease successful treatment
bull Need further studies to improve and develop laboratory diagnostic tool for IFI
CULTURE
bull Not sensitive interpreted with caution (contamination)
bull Recovered from blood LCS
bull Aspergillus isolated from BALsputum obtained from patients with a pulmonary infiltrates indicative of infection
bull A fumigatus can be isolated from sputum of patients suffer from allergic bronchopulmonary aspergillosis
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
SEROLOGICAL TESTS
bull Antobodies againts Aspergillus often helpful in the diagnosis of aspergillosis in non immunocompromised patients
bull Immunodiffusion indirect haemagglutination ELISA
bull The role is still uncertain
bull Positive ID test is not a proof of infection negative result can not exclude the diagnosis
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
SEROLOGICAL TESTS
bull Antigen detection (galactomannan) in blood and other body fluids rapid diagnosis of aspergilllosis in immunocompromised patients
bull Galactomannan fluctuate during the course of the infection useful if performed in a regular basis (twice per week)
bull Negative not exclude the diagnosis
bull ELISA for the detection of galactomannan has been approved for diagnostic use (the accuracy was moderate)
Christopher D Pfeiffer et al 2006
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
MOLECULAR DIAGNOSTICS
bull PCR for detection of Aspergillus in blood BAL serum etc
bull Appear promising for rapid detection of Aspergillus directly on tissue or body fluids but
ndash False positive
ndash No standardized commercial method
bull Routine use of PCR can not be recommended
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
Mark Reinwald et al
bull ldquoTreatment with mould-active antifungals prior to BAL sampling significantly decreases the performance of the Aspergillus PCR assay in haematological patients if BAL was performed after administration of more than one antifungal agentrdquo
bull ldquoThe effect of antifungal therapy on other surrogate parameters suggests reduced sensitivity rates with antifungal therapy for GM whereas no influence of antifungal therapy on β-D-glucan performance in a (single-centre retrospective) study was describedrdquo
J Antimicrob Chemother 2012 67 2260ndash2267
CONCLUSION
bull Various detection methods of agents responsible for invasive fungal infections have been developed
bull The lack of sensitivity and specificity of the test uncertainty in diagnosing IFI decrease successful treatment
bull Need further studies to improve and develop laboratory diagnostic tool for IFI
SEROLOGICAL TESTS
bull Antobodies againts Aspergillus often helpful in the diagnosis of aspergillosis in non immunocompromised patients
bull Immunodiffusion indirect haemagglutination ELISA
bull The role is still uncertain
bull Positive ID test is not a proof of infection negative result can not exclude the diagnosis
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
SEROLOGICAL TESTS
bull Antigen detection (galactomannan) in blood and other body fluids rapid diagnosis of aspergilllosis in immunocompromised patients
bull Galactomannan fluctuate during the course of the infection useful if performed in a regular basis (twice per week)
bull Negative not exclude the diagnosis
bull ELISA for the detection of galactomannan has been approved for diagnostic use (the accuracy was moderate)
Christopher D Pfeiffer et al 2006
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
MOLECULAR DIAGNOSTICS
bull PCR for detection of Aspergillus in blood BAL serum etc
bull Appear promising for rapid detection of Aspergillus directly on tissue or body fluids but
ndash False positive
ndash No standardized commercial method
bull Routine use of PCR can not be recommended
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
Mark Reinwald et al
bull ldquoTreatment with mould-active antifungals prior to BAL sampling significantly decreases the performance of the Aspergillus PCR assay in haematological patients if BAL was performed after administration of more than one antifungal agentrdquo
bull ldquoThe effect of antifungal therapy on other surrogate parameters suggests reduced sensitivity rates with antifungal therapy for GM whereas no influence of antifungal therapy on β-D-glucan performance in a (single-centre retrospective) study was describedrdquo
J Antimicrob Chemother 2012 67 2260ndash2267
CONCLUSION
bull Various detection methods of agents responsible for invasive fungal infections have been developed
bull The lack of sensitivity and specificity of the test uncertainty in diagnosing IFI decrease successful treatment
bull Need further studies to improve and develop laboratory diagnostic tool for IFI
SEROLOGICAL TESTS
bull Antigen detection (galactomannan) in blood and other body fluids rapid diagnosis of aspergilllosis in immunocompromised patients
bull Galactomannan fluctuate during the course of the infection useful if performed in a regular basis (twice per week)
bull Negative not exclude the diagnosis
bull ELISA for the detection of galactomannan has been approved for diagnostic use (the accuracy was moderate)
Christopher D Pfeiffer et al 2006
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
MOLECULAR DIAGNOSTICS
bull PCR for detection of Aspergillus in blood BAL serum etc
bull Appear promising for rapid detection of Aspergillus directly on tissue or body fluids but
ndash False positive
ndash No standardized commercial method
bull Routine use of PCR can not be recommended
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
Mark Reinwald et al
bull ldquoTreatment with mould-active antifungals prior to BAL sampling significantly decreases the performance of the Aspergillus PCR assay in haematological patients if BAL was performed after administration of more than one antifungal agentrdquo
bull ldquoThe effect of antifungal therapy on other surrogate parameters suggests reduced sensitivity rates with antifungal therapy for GM whereas no influence of antifungal therapy on β-D-glucan performance in a (single-centre retrospective) study was describedrdquo
J Antimicrob Chemother 2012 67 2260ndash2267
CONCLUSION
bull Various detection methods of agents responsible for invasive fungal infections have been developed
bull The lack of sensitivity and specificity of the test uncertainty in diagnosing IFI decrease successful treatment
bull Need further studies to improve and develop laboratory diagnostic tool for IFI
MOLECULAR DIAGNOSTICS
bull PCR for detection of Aspergillus in blood BAL serum etc
bull Appear promising for rapid detection of Aspergillus directly on tissue or body fluids but
ndash False positive
ndash No standardized commercial method
bull Routine use of PCR can not be recommended
Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management
Mark Reinwald et al
bull ldquoTreatment with mould-active antifungals prior to BAL sampling significantly decreases the performance of the Aspergillus PCR assay in haematological patients if BAL was performed after administration of more than one antifungal agentrdquo
bull ldquoThe effect of antifungal therapy on other surrogate parameters suggests reduced sensitivity rates with antifungal therapy for GM whereas no influence of antifungal therapy on β-D-glucan performance in a (single-centre retrospective) study was describedrdquo
J Antimicrob Chemother 2012 67 2260ndash2267
CONCLUSION
bull Various detection methods of agents responsible for invasive fungal infections have been developed
bull The lack of sensitivity and specificity of the test uncertainty in diagnosing IFI decrease successful treatment
bull Need further studies to improve and develop laboratory diagnostic tool for IFI
Mark Reinwald et al
bull ldquoTreatment with mould-active antifungals prior to BAL sampling significantly decreases the performance of the Aspergillus PCR assay in haematological patients if BAL was performed after administration of more than one antifungal agentrdquo
bull ldquoThe effect of antifungal therapy on other surrogate parameters suggests reduced sensitivity rates with antifungal therapy for GM whereas no influence of antifungal therapy on β-D-glucan performance in a (single-centre retrospective) study was describedrdquo
J Antimicrob Chemother 2012 67 2260ndash2267
CONCLUSION
bull Various detection methods of agents responsible for invasive fungal infections have been developed
bull The lack of sensitivity and specificity of the test uncertainty in diagnosing IFI decrease successful treatment
bull Need further studies to improve and develop laboratory diagnostic tool for IFI
CONCLUSION
bull Various detection methods of agents responsible for invasive fungal infections have been developed
bull The lack of sensitivity and specificity of the test uncertainty in diagnosing IFI decrease successful treatment
bull Need further studies to improve and develop laboratory diagnostic tool for IFI