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Accredited for compliance with ISO/IEC 17043Accreditation Number: 14863
RCPAQAP Molecular Infectious DiseasesGeneric ReportMAF1:2015Molecular Alphavirus / Flavivirus
CopyrightThis material is copyright and may not be used in any form for advertising, sales promotion or publicity. The material maynot be reproduced in whole or in part for any purpose whatsoever (including presentations at meetings and conferences),without the prior written permission of the RCPA Quality Assurance Programs Pty Limited. Permission must be sought inwriting from the Program but will not be unreasonably refused.
ConfidentialityRCPA Quality Assurance Programs Pty Limited keeps all participant details confidential. No information related to any of theparticipants will be divulged to a third party, unless required by legislation, without the express written consent of theparticipant. General information may be discussed at meetings or presented as papers to journals.
RCPA Quality Phone +61 2 9045 6000Assurance Programs Fax +61 2 9356 2003
ABN 32 003 520 072Suite 201, 8 Herbert StreetSt Leonards NSW 2065 www.rcpaqap.com.au
© 2015 RCPA Quality Assurance Programs Pty Ltd. All rights reservedReport Issued Monday, 18 May 2015Page Page 2 of 28
Generic Report MAF1:2015Participant ResultsThe Molecular Alphavirus/Flavivirus Program was distributed to 14 participants.12 participants returned results for this survey.
Clerical ErrorThere was a clerical error included in this survey for Specimen 1B.
Specimen 1B labelled: Rachael SCHULT; U/N 257032Instructions to Participant labelled: Rachael SCHULTZ: U/N 257032
9/12 (75%) participants detected the clerical error for Specimen 1B.
3/12 (25%) participants (1187, 83000 and 89082) did not identify the clerical error.
Specimen DetailsSamples consist of inactivated virus (gamma inactivation radiation (50 kGy)) from human strains of alphavirus orflavivirus suspended in 250uL of tissue culture fluid or molecular grade water.(Ref. 2015 RCPAQAP Molecular Infectious Diseases Program Plan section MAF1:2015)
1A Positive for Alphavirus, Ross River virus.
1B Negative, to assess contamination.
1C Positive for Flavivirus, Dengue 3 virus.
1D Positive for Alphavirus, Chikungunya virus.
1E Positive for Flavivirus, Dengue 1 virus.
1F Positive for Flavivirus, Japanese Encephalitis.
Source: Molecular Diagnostics & Development, Division of Microbiology and Infectious Diseases, PathWest LaboratoryMedicine, WA and RCPAQAP Serology MDM Serum Bank.
Preliminary & Pre-issue testing: Molecular Diagnostics & Development, Division of Microbiology and Infectious Diseases,PathWest Laboratory Medicine, WA and Public Health Laboratory, Queensland Health Forensic and Scientific Services,QLD.
Survey AimsThe detection and identification of Alphavirus (Ross River and Chikungunya) and Flavivirus (Dengue type 1, Dengue type3 and Japanese Encephalitis). A negative is included to assess contamination.
© 2015 RCPA Quality Assurance Programs Pty Ltd. All rights reservedReport Issued Monday, 18 May 2015Page Page 3 of 28
Clinical Notes (written by Dr Ed Raby)The “Clinical Notes” were created so as to highlight particular issues relating to interpretation of results. They do notnecessarily reflect the clinical history of the patient from whom the sample was obtained.
1A 24 year old female with fever, joint pains, rash. Rubella?
1B 42 year old female with polyarthralgia, tired.
1C 18 year old male, headache, rash, returned from Fiji.
1D 64 year old male, fever, hand and ankle pains, recent travel to Bali.
1E 32 year old male, travel to Thailand, fever, thrombocytopaenia.
1F 20 year old female, holiday work on pig farm in Vietnam, polio‐like syndrome.
Pre-issue ResultsThese results were obtained in pre-issue testing of the survey specimens.
Specimen Alphavirus Result Flavivirus Result Virus Identification[RT PCR]
1A positive negative Ross River
1B negative negative not applicable
1C negative positive Dengue 3
1D positive negative Chikungunya
1E negative positive Dengue 1
1F negative positive Japanese Encephalitis
Method PerformanceIt is up to all users to form their own opinion or conclusions on each kit's performance based on the accumulation ofdata and information that is supplied by this survey, and other information available to the participant.
Please note when primer sequence information is returned by participants it is not published to protect confidentiality.
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Result Review
ALPHAVIRUS Results10 participants returned results for Alphavirus molecular analysis.
Specimen 1A is positive for an Alphavirus; Ross River virus
3/4 participants (10003, 40002 and 83000) correctly detected an Alphavirus using Alphavirus group testing.Participant 10003 further commented on the identification of Ross River virus.
Participant 89112 failed to detect an Alphavirus using specific Alphavirus group testing.
All participants (40002, 60003 and 80071) performing specific Ross River virus testing identified the Ross River virus.
Specimen 1B is negative for an Alphavirus; all participants correctly returned a negative / not detected result.
Specimen 1C is negative for an Alphavirus; all participants correctly returned a negative / not detected result.
Specimen 1D is positive for an Alphavirus; Chikungunya virus.
3/4 participants (10003, 40002 and 83000) correctly detected an Alphavirus using Alphavirus group testing.Participants 10003 and 83000 further commented on the identification of Chikungunya virus.
Participant 89112 failed to detect an Alphavirus using specific Alphavirus group testing.
All participants (10003, 40002, 60003, 80011, 80042, 80069, 80071, 83000, 89082 and 89112) performing specificChikungunya virus testing identified the Chikungunya virus.
Specimen 1E is negative for an Alphavirus; all participants correctly returned a negative / not detected result.
Specimen 1F is negative for an Alphavirus; all participants correctly returned a negative / not detected result.
The results reported by Participant 1187 in the Alphavirus section were moved to the Dengue section as a data entryerror had been made.
Table 1 Alphavirus Group5 participants returned results.
Molecular A/F Alphavirus Group
Part1A 1B 1C 1D 1E 1F
Result Comment Result Comment Result Result Comment Result Result
10003 detected typed as ross rivervirus by sequencing
notdetected
notdetected detected
typed as chikungunyavirus by real-time PCR(Ct=20) & confirmed bysequencing
notdetected
notdetected
40002 detected notdetected
notdetected detected not
detectednot
detected
83000 detected A type of Alphavirusdetected
notdetected - not
detected detected
A type of Alphavirusdetected, typed asChikungunya by real-time RT-PCR
notdetected
notdetected
89112 negThis result does notexclude a recentAlpha virus infection
negThis result does notexclude a recentAlpha virus infection
nottested neg
This result does notexclude a recent Alphavirus infection
nottested
nottested
Participant 89112 appears to have used an expired amplification kit.
Participants 40002 and 89112 have not provided inhibition control information.
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ALPHAVIRUS Results (continued)
Table 2 Barmah Forest Virus2 participants returned results.
Molecular A/F Barmah Forest Virus
Part 1A 1B 1C 1D 1E 1F
40002 not detected not detected not tested not detected not tested not tested
60003 not detected not detected not detected not detected not detected not detected
Participant 40002 has not provided inhibition control information.
Table 3 Chikungunya Virus10 participants returned results.
Molecular A/F Chikungunya Virus
Part 1A 1B 1C 1D 1E 1F
10003 not detected not detected not detected detected not detected not detected
40002 not detected not detected not tested detected not tested not tested
60003 not detected not detected not detected detected not detected not detected
80011 not detected not detected not detected detected not detected not detected
80042 not detected not detected not detected detected not detected not detected
80069 not detected not detected not detected detected not detected not detected
80071 neg neg neg pos neg neg
83000 not detected not detected not detected detected not detected not detected
89082 neg neg neg pos neg neg
89112 neg neg not tested pos not tested not tested
Participant 80069 commented ‘Chikungunya not detected’ for Specimens 1A, 1B, 1C, 1E and 1F.
Participant 89112 commented ‘This result does not exclude a recent Chikungunya virus infection’ for Specimens 1A and1B. Participant 89112 commented ‘This result suggest a recent infection with Chikungunya virus’ for Specimen 1D.
Participants 40002, 80069, 80071, 89082 and 89112 have not provided inhibition control information.
Table 4 Ross River Virus3 participants returned results.
Molecular A/F Ross River Virus
Part 1A 1B 1C 1D 1E 1F
40002 detected not detected not tested not detected not tested not tested
60003 detected not detected not detected not detected not detected not detected
80071 pos neg neg neg neg neg
Participants 40002 and 80071 have not provided inhibition control information.
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ALPHAVIRUS Results (continued)
Table 5 Sindbis Virus1 participant returned results.
Molecular A/F Sindbis Virus
Part 1A 1B 1C 1D 1E 1F
40002 not detected not detected not tested not detected not tested not tested
Participant 40002 has not provided inhibition control information.
FLAVIVIRUS Results12 participants returned results for Flavivirus molecular analysis.
Specimen 1A is negative for a Flavivirus; all participants correctly returned a negative / not detected result.
Specimen 1B is negative for a Flavivirus; most participants correctly returned a negative / not detected result.
Participant 10003 incorrectly detected a Flavivirus using Flavivirus group testing and commented ‘typed as japaneseencephalitis virus by sequencing’. This participant reported a correct result of ‘not detected’ for specific JapaneseEncephalitis virus, however commented ‘positive in Flavivirus Group assay and typed as JEV by sequencing’.
Specimen 1C is positive for a Flavivirus; Dengue 3 virus.
All participants correctly detected a Flavivirus using Flavivirus group testing.Participants 10003 and 83000 also correctly commented on the identification of Dengue virus 3.
All participants performing specific Dengue virus testing identified the Dengue virus.Participants 10003, 40002, 60003, 80042, 80069, 80071 and 83000 also correctly identified the virus as Dengue type 3.
Specimen 1D is negative for a Flavivirus; most participants correctly returned a negative / not detected result.
Participant 10003 incorrectly detected a Flavivirus using Flavivirus group testing and commented ‘typed as dengue virustype 3 by real-time PCR (Ct=36) & confirmed by sequencing’. This participant also incorrectly identified Dengue type 3,using specific Dengue testing.
Specimen 1E is positive for a Flavivirus; Dengue 1 virus.
All participants correctly detected a Flavivirus using Flavivirus group testing.Participants 10003 and 83000 also correctly commented on the identification of Dengue virus 1.
All participants performing specific Dengue virus testing identified the Dengue virus.Participants 10003, 40002, 60003, 80042, 80069, 80071 and 83000 also correctly identified the virus as Dengue type 1.
Specimen 1F is positive for a Flavivirus; Japanese Encephalitis virus.
All participants correctly detected a Flavivirus using Flavivirus group testing.Participants 10003 and 83000 also correctly commented on the identification of Japanese Encephalitis virus.
All participants performing specific Japanese Encephalitis virus testing identified the Japanese Encephalitis virus.
Participant 60003 incorrectly reported ‘detected’ for this specimen using specific West Nile Virus assay, furthercommenting ‘WN and JE POS’.
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FLAVIVIRUS Results (continued)
Table 6 Flavivirus Group5 participants returned results.
Molecular A/F Flavivirus Group
Part1A 1B 1C 1D 1E 1F
Result Result Result Comment Result Result Comment Result Comment
10003 notdetected detected detected
typed as dengue virustype 3 by real-timePCR (Ct=17) &confirmed bysequencing
detected detected
typed as dengue virustype 1 by real-timePCR (Ct=18) &confirmed bysequencing
detected
typed as japaneseencephalitis virus byreal-time PCR (Ct=27)& confirmed bysequencing
40002 notdetected
notdetected detected not
detected detected detected
80071 neg neg pos neg pos pos
83000 notdetected
notdetected detected
A type of Flavivirusdetected, typed asDengue 3 by real-timeRT-PCR
notdetected detected
A type of Flavivirusdetected, typed asDengue 1 by real-timeRT-PCR
detectedA type of Flavivirusdetected, typed as JEby real-time RT-PCR
89112 neg neg posThis result suggest arecent infection withFlavivirus
neg posThis result suggest arecent infection withFlavivirus
posThis result suggest arecent infection withFlavivirus
Participant 10003 incorrectly returned ‘detected’ for Specimens 1B and 1D and further commented ‘typed as japaneseencephalitis virus by sequencing’ and ‘typed as dengue virus type 3 by real-time PCR (Ct=36) & confirmed by sequencing’respectively.
Participant 89112 commented ‘This result does not exclude a recent Flavivirus infection’ for Specimens 1A, 1B and 1D.
Participants 40002, 80071 and 89112 have not provided inhibition control information.
Table 7 Dengue Virus12 participants returned results.
Molecular A/F Dengue Virus
Part1A 1B 1C 1D 1E 1F
Result Result Result Comment Result Result Comment Result
1187 notdetected
notdetected detected not
detected detected notdetected
10003 notdetected
notdetected detected dengue virus type 3 detected detected detected dengue virus type 1
detectednot
detected
40002 nottested
notdetected detected Dengue 3 RNA was detected in
this samplenot
tested detected Dengue 1 RNA was detectedin this sample
notdetected
60003 notdetected
notdetected detected Dengue Type 3 POS not
detected detected Dengue Type 1 POS notdetected
80011 notdetected
notdetected detected not
detected detected notdetected
80042 notdetected
notdetected detected Dengue Type 3 not
detected detected Dengue Type 1 notdetected
80052 neg neg pos neg pos neg
80069 notdetected
notdetected detected DEN 3 Detected not
detected detected DEN 1 Detected notdetected
80071 neg neg pos D3 neg pos D1 neg
83000 notdetected
notdetected detected Dengue type 3 detected not
detected detected Dengue type 1 detected notdetected
89082 neg neg pos neg pos neg
89112 nottested
nottested pos This result suggest a recent
infection with Dengue virusnot
tested pos This result suggest a recentinfection with Dengue virus neg
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FLAVIVIRUS Results (continued)
Participant 1187 had their results moved from the Alphavirus section to the Dengue section based on the kit theyreported using.
Participant 10003 incorrectly returned ‘detected’ for Specimen 1D and further commented ‘dengue virus type 3detected’.
Participants 1187, 40002, 80042, 80069, 80071 and 89082 have not provided inhibition control information.
Participant 1187 has not provided negative or positive control information.
Participant 89112 appears to have used an expired amplification and detection kit.
Table 8 Japanese Encephalitis Virus7 participants returned results.
Molecular A/F Japanese Encephalitis Virus
Part1A 1B 1C 1D 1E 1F
Result Result Result Result Result Result Comment
10003 not detected not detected not detected not detected not detected detected
40002 not tested not detected not detected not tested not detected detected
60003 not detected not detected not detected not detected not detected detected
80069 not detected not detected not tested not tested not tested detected Japanese Encephalitisvirus Detected
80071 neg neg neg neg neg pos
83000 not detected not detected not detected not detected not detected detected JE Virus detected
89082 neg neg neg neg neg pos
Participant 10003 has incorrectly commented ‘positive in Flavivirus Group assay and typed as JEV by sequencing’ forSpecimen 1B.
Participant 60003 has not provided master/sample volume for the assay in use.
Participants 40002, 80069, 80071 and 89082 have not provided inhibition control information.
Table 9 Kunjin Virus2 participants returned results.
Molecular A/F Kunjin Virus
Part 1A 1B 1C 1D 1E 1F
40002 not tested not detected not detected not tested not detected not detected
60003 not detected not detected not detected not detected not detected not detected
Participant 40002 has not provided inhibition control information.
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FLAVIVIRUS Results (continued)
Table 10 Murray Valley Encephalitis Virus3 participants returned results.
Molecular A/F Murray Valley Encephalitis Virus
Part 1A 1B 1C 1D 1E 1F
40002 not tested not detected not detected not tested not detected not detected
60003 not detected not detected not detected not detected not detected not detected
80069 not detected not detected not tested not tested not tested not detected
Participants 40002 and 80069 have not provided inhibition control information.
Table 11 West Nile Virus5 participants returned results.
Molecular A/F West Nile Virus
Part 1A 1B 1C 1D 1E 1F
40002 not tested not detected not detected not tested not detected not detected
60003 not detected not detected not detected not detected not detected detected
80069 not detected not detected not tested not tested not tested not detected
89082 neg neg neg neg neg neg
89112 neg neg neg neg neg neg
Participant 60003 incorrectly returned ‘detected’ for Specimen 1F further commented ‘WN and JE POS’.
Participant 60003 has not provided master/sample volumes for the assay in use.
Participants 40002, 80069, 89082 and 89112 have not provided inhibition control information.
Participant 40002 has not provided negative control information.
Table 12 Yellow Fever Virus5 participants returned results.
Molecular A/F Yellow Fever Virus
Part 1A 1B 1C 1D 1E 1F
40002 not tested not detected not detected not tested not detected not detected
80069 not detected not detected not tested not tested not tested not detected
80071 neg neg neg neg neg neg
89082 neg neg neg neg neg neg
89112 neg neg neg neg neg neg
Participants 40002, 80069, 80071, 89082 and 89112 have not provided inhibition control information.
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Review
Clinical Notes [Discussion by Dr Ed Raby, PathWest, WA]The “Clinical Notes” were created so as to highlight particular issues relating to interpretation of results. They do notnecessarily reflect the clinical history of the patient from whom the sample was obtained.
1A 24 year old female with fever, joint pains, rash. Rubella?Typical symptoms of RRV include a rash in about 50% of patients that may be clinically indistinguishable fromthe rash of rubella.
1B 42 year old female with polyarthralgia, tired.
1C 18 year old male, headache, rash, returned from Fiji.Fiji has had more than 25,000 suspected cases of Dengue with 15 deaths and remains an important source ofimported cases to Queensland.
1D 64 year old male, fever, hand and ankle pains, recent travel to Bali.Chikungunya is now endemic in Bali and may be difficult to distinguish clinically from Dengue Fever. The jointpain/arthralgia of CHIK infection typically affects the distal joints, similar to RRV/BFV and in contrast to Denguewhich is typically more central.
1E 32 year old male, travel to Thailand, fever, thrombocytopaenia.Low platelet count (thrombocytopaenia) is a common finding in Dengue Fever although significant haemorrhageremains very rare in adult travellers.
1F 20 year old female, holiday work on pig farm in Vietnam, polio‐like syndrome.Japanese Encephalitis Virus is present throughout Asia and causes epidemics and seasonal disease. It is rare intravellers without prolonged rural exposure. Polio‐like flaccid paralysis as well as more generalised encephalitisoccur in a minority of patients with most infections remaining asymptomatic.
AlphavirusSpecimens 1A and 1D were positive for an Alphavirus.
Four participants submitted group results. Three participants correctly detected the presence of an alphavirus in boththe above specimens. Participant 89112 failed to detected alphavirus in either Specimen 1A or 1D despite successfuldetection of both Ross River virus and Chikungunya virus in previous surveys using a pan-alpha PCR.
Three participants returning alphavirus results identified Ross River virus in Specimen 1A using a specific PCR.
Ten participants returning alphavirus results identified Chikungunya virus in Specimen 1D using a specific PCR.
There were no false positive results.
FlavivirusSpecimens 1C, 1E and 1F were positive for a Flavivirus.
Five participants submitted group results. All participants correctly detected the presence of a flavivirus in the abovespecimens. False positive results reported in Specimens 1B and 1D should be further investigated by Participant 10003.This may represent clerical error, however appears most likely to be due to cross-contamination of specimens duringprocessing.
Twelve participants returned Dengue virus specific assay results and although all were correctly detected for Specimens1C and 1E, only seven were able to go on and provide typing results.
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Japanese encephalitis virus (Specimens 1F) was detected by all seven participants using a specific PCR. The mixeddetection reported by Participant 60003 (West Nile virus) has been reported in previous surveys as ‘cross reactivity’ andmost likely relates to primer specificity.
General CommentsIt is noted that the terminology used by several participants is incorrect, inconsistent and/or incomplete. A standardisedapproach to data returned to the RCPAQAP is encouraged. Generally in-house manufactured controls, amplification anddetection systems are poorly defined. Data entry errors are also noted. The general use of a Ct value of 0 to define anegative is not informative and is technically incorrect. Some indication of the upper and lower thresholds for each assaywould be more useful information. i.e. NEG = Ct >45 ; POS = Ct <35.
Participant 60003 reported using 160uL of specimen in the QIAamp Viral RNA Mini Kit. This is a non-standard specimenvolume for this purification method. The same participant reported the use of a 12uL volume of nucleic acid with 8uL ofreaction mix. These results are contradictory to previous submissions and may be clerical errors.
Participant 89112 used an expired Dengue virus kit. Although this did not seem to affect detection of these surveyspecimens, it may not perform as efficiently on specimens nearer to the lower limit of detection of the assay.
All participants are encouraged to incorporate an inhibition control into each assay and consider the development of apan-alpha / pan-flavi approach where possible.
There has been a large increase in the number of RRV cases reported to the Centre for Communicable DiseaseIntelligence from both NSW and QLD in the year to date and Dengue virus detections in returned traveller’s remainshigh. The use of nucleic acid amplification detection techniques in the detection of arboviruses should be limited to thetransient acute phase of infection and typing for epidemiology. All molecular results should be considered in conjunctionwith IgM and IgG serological results
Comments prepared for and on behalf of RCPAQAP by Ms Joanne Montgomery, PathWest Laboratory Medicine,Western Australia.
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Alphavirus Controls & In house Methods for Survey MAF1:2015:1A, 1B, 1C, 1D, 1E & 1F
Alphavirus Group - Controls & In house Methods
PartInhib Control Neg Control Pos Control In-house Methods
Manufacturer Lot No. Expiry Manufacturer Lot No. Expiry Manufacturer Lot No. Expiry In-house Method Detection Method
10003 In house - BVDVspiked into sample NFW Chikungunya virus cDNA
heminestedAlphavirus-genericassay
gel electrophoresis
40002 N/A In House NA In House synthetic 26022015 NA
83000 In-house Non Template Control Positive ChikungunyaIsolate
Selected humangene as inhibitorcontrol for PCR
89112 - - NA Promega (nucleasefree water) 14748722 Jan
2016In house (ChikungunyaRNA as Positive Control) NA NA Conventional Nested
PCRAgarose gelelectrophoresis and
Barmah Forest Virus - Controls & In house Methods
PartInhib Control Neg Control Pos Control In-house Methods
Manufacturer Manufacturer Expiry Manufacturer Lot No. Expiry In-house Method Detection Method
40002 N/A in house NA Primer control, Probe control 05022015,05022015 NA Taqman RT PCR Probe fluorescence
60003 In-house In-house In-house Yes Realtime PCR
Sindbis Virus - Controls & In house Methods
Part
Inhib Control Neg Control Pos Control In-house Methods
Manufacturer Manufacturer Expiry Manufacturer Lot No. Expiry In-house Method Detection Method
40002 N/A in house NA Primer Control, Probe Control 21052013,21052013 NA Taqman RT-PCR Probe Fluorescence
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Alphavirus Controls & In house Methods for Survey MAF1:2015:1A, 1B, 1C, 1D, 1E & 1F (continued)
Chikungunya Virus - Controls & In house Methods
PartInhib Control Neg Control Pos Control In-house Methods
Manufacturer Lot No. Expiry Manufacturer Lot No. Expiry Manufacturer Lot No. Expiry In-house Method Detection Method
10003 In house - BVDVspiked into sample NFW Chikungunya virus cDNA real-time Chikungunya
specific TaqMan assay Real-time PCR
40002 N/A in house NA Primer Control, ProbeControl
23122014 ,05022015 NA Taqman RT-PCR Probe Fluorescence
60003 In-house In-house Reference Strain Yes Realtime PCR80011 In-house NA NA In-house NA NA In-house NA NA
80042 In-house In-house In-house Adapted frompublished method Real-time PCR
80069 NA NA NA In-house NA NA In-house NA NA - -
80071 NA NA NA Water NA NAIn house (CHIKV celllysate RNA as PositiveControl)
NA NA Adapted from Hasebeet al., 2002
LightCycler RNA MasterSYBR Green I
83000 In-house - NA Non TemplateControl - NA Positive Chikungunya
Isolate - NASelected human geneas inhibitor control forPCR
-
89082 In house Mar 15 Mar 16 In house Mar 15 Mar 16
89112 NA Promega (nucleasefree water) 14748722 Jan 2016 In house (CHIK RNA as
Positive Control NA NA Taqman RT-PCR LightCycler
Fluorescence/LightCycler
Ross River Virus - Controls & In house Methods
PartInhib Control Neg Control Pos Control In-house Methods
Manufacturer Lot No. Expiry Manufacturer Lot No. Expiry Manufacturer Lot No. Expiry In-house Method Detection Method
40002 N/A in house NA Primer control, Probe Control 10032015,10032015 NA Taqman RT-PCR Probe Fluorescence
60003 In-house In-house Reference strain Yes Realtime PCR
80071 NA NA NA Water NA NA In house (Ross river virus celllysate RNA as Positive Control) NA NA Adapted from
Studdert et al., 2003LightCycler® RNAMaster SYBR Green I
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Alphavirus Units and Cut-Off Values for Survey MAF1:2015:1A, 1B, 1C, 1D, 1E & 1F
Part UnitsAlphavirus Group - Units & Cut Off
Neg Equ Pos
10003 PCR product size no specific electrophoresis sizeband/IC OK
specific electrophoresis sizeband/IC OK
40002 band intesity83000 MELT CURVE NO MELT CURVE - MELT CURVE DETECTED89112 No units available
Part Units
Barmah Forest Virus - Units & Cut Off
Neg Equ Pos
40002 Ct 40 37-39 3660003 Ct 0 39-40 1-39
Part Units
Chikungunya Virus - Units & Cut Off
Neg Equ Pos
10003 Ct no specific amplification/IC OK specific amplification/IC OK40002 Ct 40 37-39 3660003 Ct 0 39-40 1-3980011 Ct80042 Cp 50 45-50 4580069 Ct 0.00 & above 40.00 - Below 40.0
80071 Cp No product formation at meltingcurve analysis
Product formation at meltingcurve analysis
83000 Ct Below Ct value - Above Ct value89082 Ct 45 40 2989112 No units available
Part UnitsRoss River Virus - Units & Cut Off
Neg Equ Pos
40002 Ct 40 37-39 3660003 Ct 0 39-40 1-39
80071 Cp No product formation atmelting curve analysis
Product formation at meltingcurve analysis
Part Units
Sindbis Virus - Units & Cut Off
Neg Equ Pos
40002 Ct 40 37-39 36
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Alphavirus Extraction Kit Information for Survey MAF1:2015:1A, 1B, 1C, 1D, 1E & 1F
PartAlphavirus Group - Extraction
Manufacturer Kit Name Lot No. Expiry Date Extraction Vol Elution Vol
10003 Qiagen QIAamp Viral RNA Mini Kit 148026903 Apr 16 140 6040002 Qiagen EZ1 Virus Mini Kit v2.0 148038536 Mar 16 200 9083000 Qiagen QIAamp Viral RNA Mini Kit 148017918 Jan 16 140 5089112 Qiagen QIAamp Viral RNA Mini Kit 148013001 Nov 15 140 60
Part
Barmah Forest Virus - Extraction
Manufacturer Kit Name Lot No. Expiry Date Extraction Vol Elution Vol
40002 Qiagen EZ1 Virus Mini Kit v2.0 148038536 Mar 16 200 9060003 Qiagen QIAamp Viral RNA Mini Kit 148044563 Aug 16 160 100
Part
Chikungunya Virus - Extraction
Manufacturer Kit Name Automation Lot No. Expiry Date ExtractionVol
ElutionVol
10003 Qiagen QIAamp Viral RNA Mini Kit - 148026903 Apr 16 140 6040002 Qiagen EZ1 Virus Mini Kit v2.0 - 148038536 Mar 16 200 9060003 Qiagen QIAamp Viral RNA Mini Kit - 148044563 Aug 16 160 10080011 bioMerieux NucliSens EasyMag Z015NA Dec 15 200 2580042 Qiagen EZ1 Virus Mini Kit v2.0 - 148046999 Nov 15 200 6080069 Qiagen QIAamp Viral RNA Mini Kit - 148045591 Sep 16 140 6080071 Qiagen QIAamp Viral RNA Mini Kit - 148052920 Oct 16 140 6083000 Qiagen QIAamp Viral RNA Mini Kit - 148017918 Jan 16 140 50
89082 Roche Diagnostics MagNA Pure Compact NucleicAcid Isolation Kit I MagNA Pure 10771500 Dec 15 200 50
89112 Qiagen QIAamp Viral RNA Mini Kit - 148013001 Nov 15 140 60
Part
Ross River Virus - Extraction
Manufacturer Kit Name Lot No. Expiry Date Extraction Vol Elution Vol
40002 Qiagen EZ1 Virus Mini Kit v2.0 148038536 Mar 16 200 9060003 Qiagen QIAamp Viral RNA Mini Kit 148044563 Aug 16 160 10080071 Qiagen QIAamp Viral RNA Mini Kit 148052920 Oct 16 140 60
Part
Sindbis Virus - Extraction
Manufacturer Kit Name Lot No. Expiry Date Extraction Vol Elution Vol
40002 Qiagen EZ1 Virus Mini Kit v2.0 148038536 Mar 16 200 90
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Alphavirus Amplification Kit Information for Survey MAF1:2015:1A, 1B, 1C, 1D, 1E & 1F
Part
Alphavirus - Amplification
Manufacturer Kit Name Method Automation Lot No. ExpiryDate
MasterVol
SampleVol
Single/Nested
10003 In house - RT PCR Eppendorf GradientS Thermalcycler NA NA 37 3 Nested
40002 In house - RT PCR Eppendorf GradientS Thermalcycler 19032015 Mar 16 15 5 Nested
83000 BioRad iScript One-StepRT-PCR iQ5 170-8893 Dec 15 20 5 Single round
89112 Roche AppliedScience
Expand HighFidelity PLUS
AB 2720 ThermalCycler 13434025 Mar 14 48.8 1 Nested
Part
Barmah Forest Virus - Amplification
Manufacturer Kit Name Method Automation Lot No. ExpiryDate
MasterVol
SampleVol Single/Nested
40002 In house - TaqMan RT-PCR Rotorgene 20052014 May 15 15 5 Single round60003 In house - TaqMan RT-PCR Rotorgene NA NA 8 12 Single round
Part
Chikungunya Virus - Amplification
Manufacturer Kit Name Method Automation Lot No. ExpiryDate
MasterVol
SampleVol
Single/Nested
10003 In house - TaqMan RT-PCR ABI Prism NA NA 17 3 Single round40002 In house - TaqMan RT-PCR Rotorgene 04032015 Mar 16 15 5 Single round60003 In house - TaqMan RT-PCR Rotorgene NA NA 12 8 Single round
80011 Invitrogen SuperScript III qRT-PCR RT PCR Light Cycler 1565698 Apr 16 15 5 Single round
80042 In house - Real-time PCR Light Cycler NA NA 20 5 Single round80069 AITbiotech abTES DEN 5 qPCR RT-PCR CFX96 C151-24 May 15 20 5 Single round80071 In house - Real-time PCR Light Cycler 10035000 Sep 15 9 1 Single round83000 Qiagen OneStep RT-PCR Kit RT-PCR - 148030425 Jul 15 20 5 Single round89082 In house - TaqMan RT-PCR LightCycler 110315 Mar 16 15 5 Single round
89112 RocheDiagnostics
LightCycler RNAAmplification kitHypro
RT-PCR Light Cycler 14748722 Jan 16 15 5 Single round
Part
Ross River Virus - Amplification
Manufacturer Kit Name Method Automation Lot No. ExpiryDate
MasterVol
SampleVol Single/Nested
40002 In house - TaqMan RT-PCR Rotorgene 05022015 Feb 16 15 5 Single round60003 In house - TaqMan RT-PCR Rotorgene NA NA 12 8 Single round80071 In house - Real-time PCR Light Cycler 10035000 Sep 15 9 1 Single round
Part
Sindbis Virus - Amplification
Manufacturer Kit Name Method Automation Lot No. ExpiryDate
MasterVol
SampleVol Single/Nested
40002 In house - TaqMan RT-PCR Rotorgene 21012015 Jan 16 15 5 Single round
© 2015 RCPA Quality Assurance Programs Pty Ltd. All rights reservedReport Issued Monday, 18 May 2015Page Page 17 of 28
Alphavirus Detection Kit Information for Survey MAF1:2015:1A, 1B, 1C, 1D, 1E & 1F
Part
Alphavirus Group - Detection
Manufacturer Kit Name Method Automation Lot No. ExpiryDate Single/Nested
10003 In house - Gelelectrophoresis - NA NA Nested
40002 Qiagen QIAxcel DNA HighResolution Kit QIAxcel C131031033 May 15 Nested
83000 BioRad iScript One-Step RT-PCR iQ5 170-8893 Dec 15 Single round
89112 UV Products UV Transilluminator GeneSnap fromSynGene IN HOUSE Oct 15 Nested
Part
Barmah Forest Virus - Detection
Manufacturer Kit Name Method Automation Lot No. ExpiryDate Single/Nested
40002 In house - TaqMan RT-PCR Rotorgene 20052014 May 15 Single round60003 In house - TaqMan RT-PCR Rotorgene NA NA Single round
Part
Chikungunya Virus - Detection
Manufacturer Kit Name Method Automation Lot No. ExpiryDate Single/Nested
10003 In house - TaqMan RT-PCR ABI Prism NA NA Single round40002 In house - TaqMan RT-PCR Rotorgene 04032015 Mar 16 Single round60003 In house - TaqMan RT-PCR Rotorgene NA NA Nested80011 In house - TaqMan RT-PCR LightCycler 1565698 Apr 16 Single round80042 In house - Real-time PCR Light Cycler NA NA Single round80069 AITbiotech abTES DEN 5 qPCR RT-PCR CFX96 C151-24 May 15 Single round80071 In house - Real-time PCR Light Cycler 10035000 Sep 15 Single round
83000 Qiagen QuantiTect Probe RT-PCR Kit
Fluorescent(FAM channel) - 148030425 Jul 15 Single round
89082 In house - TaqMan RT-PCR LightCycler 110315 Mar 16 Single round89112 In house - TaqMan RT-PCR LightCycler IN HOUSE Jan 16 Single round
Part
Ross River Virus - Detection
Manufacturer Kit Name Method Automation Lot No. ExpiryDate Single/Nested
40002 In house - TaqMan RT-PCR Rotorgene 05022015 Feb 16 Single round60003 In house - TaqMan RT-PCR Rotorgene NA NA Single round80071 In house - Real-time PCR Light Cycler 10035000 Sep 15 Single round
PartSindbis Virus - Detection
Manufacturer Kit Name Method Automation Lot No. Expiry Date Single/Nested
40002 In house - TaqMan RT-PCR Rotorgene 21012015 Jan 16 Single round
© 2015 RCPA Quality Assurance Programs Pty Ltd. All rights reservedReport Issued Monday, 18 May 2015Page Page 18 of 28
Flavivirus Controls & In house Methods for Survey MAF1:2015:1A, 1B, 1C, 1D, 1E & 1F
Flavivirus Group - Controls & In house Methods
PartInhib Control Neg Control Pos Control In-house Methods
Manufacturer Lot No. Expiry Manufacturer Lot No. Expiry Manufacturer Lot No. Expiry In-house Method Detection Method
10003 In house - BVDVspiked into sample NFW MVE cDNA Flavivirus generic
PCR gel electrophoresis
40002 N/A in house NA in house synthetic 28082014 NA Nested RT-PCR band on ethidiumbromide gel
80071 NA NA NA Water NA NA In house (JEV cell lysateRNA as Positive Control) NA NA Adapted from
Moureau et al, 2007QuantiFast SYBRgreen RT-PCR
83000 In-house - NA Non TemplateControl - NA Dengue & JE Virus
Isolates - NASelected humangene as inhibitorcontrol for PCR
-
89112 - - NA Promega (nucleasefree water) 14748722 Jan 2016 In house YF RNA as
pos control NA Conventional RT-PCR
Agarose gelelectrophoresis
Japanese Encephalitis Virus - Controls & In house Methods
PartInhib Control Neg Control Pos Control In-house Methods
Manufacturer Lot No. Expiry Manufacturer Lot No. Expiry Manufacturer Lot No. Expiry In-house Method Detection Method
10003 In house - BVDVspiked into sample NFW Japanese encephalitis
virus cDNAreal-time JEV specificTaqMan assay Real-time PCR
40002 N/A in house NA Primer Control, ProbeControl
16012015,16012015 NA Taqman RT-PCR Probe Fluorescence
60003 In-house In-house Reference strain Yes Realtime PCR80069 NA NA NA In house NA NA In house NA NA Conventional RT-PCR Gel Electrophoresis
80071 NA NA NA WATER NA NA In house (JEV cell lysateRNA as Positive Control) NA NA Adapted from S. R.
Santhosh et al. 2007LightCycler® RNAMaster SYBR Green I
83000 In-house - NA Non TemplateControl - NA JE Synthetic
Oligonucleotide - NASelected humangene as inhibitorcontrol for PCR
-
89082 In house Mar 15 Mar 16 In house Mar 15 Mar 16
© 2015 RCPA Quality Assurance Programs Pty Ltd. All rights reservedReport Issued Monday, 18 May 2015Page Page 19 of 28
Flavivirus Controls & In house Methods for Survey MAF1:2015:1A, 1B, 1C, 1D, 1E & 1F (continued)
Dengue Virus - Controls & In house Methods
PartInhib Control Neg Control Pos Control In-house Methods
Manufacturer Lot No. Expiry Manufacturer Lot No. Expiry Manufacturer Lot No. Expiry In-house Method Detection Method
1187
10003 In house - BVDVspiked into sample NFW Dengue virus types 1-4
cDNAIndividual Dengue1-4 TaqMan assays Real-time PCR
40002 N/A in house NA Primer Control, ProbeControl
02032015,02032015 NA Taqman RT-PCR Probe
Fluorescence60003 In-house In-house Reference strain Yes Realtime PCR80011 In-house NA NA In-house NA NA In-house NA NA
80042 N/A In-house In-house Adapted frompublished method Real time PCR
80052 In-house NA In-house NA In-house DDCtrl-18 NA TaqMan Real TimeMultiplex RT-PCR
TaqMan Real TimeMultiplex RT-PCR
80069 NA NA NA In-house NA NA In-house NA NA - -
80071 NA NA NA Water NA NAIn house (Dengue virusserotypes 1-4 cell lysateRNA as Positive Control)
NA NA Lai et al 2007 LightCycler RNAMaster Hyb Probe
83000 In-house - NA Non TemplateControl - NA Positive DEN 1,2,3,4
Isolates - NASelected humangene as inhibitorcontrol for PCR
-
89082 In-house Mar 15 Mar 16 In-house Mar 15 Mar 16
89112 Altona IC022031401 May 16 Promega (nucleasefree water) 14748722 Jan 2016 Altona C1410 Jan 15
Murray Valley Encephalitis Virus - Controls & In house Methods
PartInhib Control Neg Control Pos Control In-house Methods
Manufacturer Lot No. Expiry Manufacturer Lot No. Expiry Manufacturer Lot No. Expiry In-house Method Detection Method
40002 N/A in house NA Probe Control, Primer Control 16012015,16012015 NA Taqman RT-PCR Probe fluorescence
60003 In-house In-house Reference strain Yes Realtime PCR80069 NA NA NA In house NA NA In house NA NA Conventional RT-PCR Gel electrophoresis
© 2015 RCPA Quality Assurance Programs Pty Ltd. All rights reservedReport Issued Monday, 18 May 2015Page Page 20 of 28
Flavivirus Controls & In house Methods for Survey MAF1:2015:1A, 1B, 1C, 1D, 1E & 1F (continued)
Kunjin Virus - Controls & In house Methods
PartInhib Control Neg Control Pos Control In-house Methods
Manufacturer Manufacturer Expiry Manufacturer Lot No. Expiry In-house Method Detection Method
40002 N/A in house NA Probe Control, Primer Control 23072014 ,16012015 NA Taqman RT-PCR Probe Fluorescence60003 In-house In-house Reference strain Yes Realtime PCR
West Nile Virus - Controls & In house Methods
PartInhib Control Neg Control Pos Control In-house Methods
Manufacturer Lot No. Expiry Manufacturer Lot No. Expiry Manufacturer Lot No. Expiry In-house Method Detection Method
40002 Probe Control; Primer Control 22072010,12032013 NA TaqMan RT-PCR Probe Flourescence
60003 In-house In-house Reference strain Yes Realtime PCR80069 NA NA NA In house NA NA In house NA NA Conventional RT-PCR Gel electrophoresis89082 In house Mar 15 Mar 16 In house Mar 15 Mar 16
89112 NA Promega (nucleasefree water) 14748722 Jan 2016 In house (WNV RNA as Positive
Control) WN 2651 TC1 NA NA Conventional PCR(Roche Titan one step)
Agarose gelelectrophoresis
Yellow Fever Virus - Controls & In house Methods
PartInhib Control Neg Control Pos Control In-house Methods
Manufacturer Lot No. Expiry Manufacturer Lot No. Expiry Manufacturer Lot No. Expiry In-house Method Detection Method
40002 N/A Inhouse Neg Control NA Primer Control, Probe Control 12122013,12122013 NA Taqman RT-PCR Probe Flourescence
80069 NA NA NA In house NA NA In house NA NA Conventional RT-PCR Gel Electrophoresis
80071 NA NA NA Water NA NA In house (Yellow fever virus celllysate RNA as Positive Control) NA NA Designed in-house
RT-PCR protocolLightCycler® RNAMaster SYBR Green I
89082 In house Mar 15 Mar 16 In house Mar 15 Mar 16
89112 NA Promega (nucleasefree water) 14748722 Jan 2016 In house YFV (RNA) as positive
control YF20/11/121 NA NA Conventional RT-PCR(Roche Titan one step)
Agarose gelelectrophoresis
© 2015 RCPA Quality Assurance Programs Pty Ltd. All rights reservedReport Issued Monday, 18 May 2015Page Page 21 of 28
Flavivirus Units and Cut-Off Values for Survey MAF1:2015:1A, 1B, 1C, 1D, 1E & 1F
Part UnitsFlavivirus Group - Units & Cut Off Values
Neg Equ Pos
10003 PCR product size no specific electrophoresis sizeband/IC OK
specific electrophoresis sizeband/IC OK
40002 band intensity
80071 Cp No product formation at meltingcurve analysis
Product formation at meltingcurve analysis
83000 Ct No band seen - Band seen89112 No units available
Part UnitsDengue Virus - Units & Cut Off Values
Neg Equ Pos
1187 Ct 40 and below - Less than 4010003 Ct no specific amplification/IC OK specific amplification/IC OK40002 Ct 40 37-39 3660003 Ct 0 39-40 1-3980011 Ct80042 Ct 45 40-45 1-4080052 Ct 39 37-39 3780069 Ct 0.00 & above 40.00 - Below 40.0
80071 Cp No amplification is detected atabs.quantification
Amplification is detected atabs.quantification
83000 Ct Below Ct value - Above Ct value89082 Ct 45 40 2989112 No units available
Part UnitsJapanese Encephalitis Virus - Units & Cut Off Values
Neg Equ Pos
10003 Ct no specific amplification/IC OK specific amplification/IC OK40002 Ct 40 37-39 3660003 Ct 0 39-40 1-3980069 - No specific band seen - Specific band seen
80071 Cp No product formation at meltingcurve analysis
Product formation at meltingcurve analysis
83000 Ct Below Ct value - Above Ct value89082 Ct 45 40 29
Part UnitsKunjin Virus - Units & Cut Off Values
Neg Equ Pos
40002 Ct 40 37-39 3660003 Ct 0 39-40 1-39
© 2015 RCPA Quality Assurance Programs Pty Ltd. All rights reservedReport Issued Monday, 18 May 2015Page Page 22 of 28
Flavivirus Units and Cut-Off Values for Survey MAF1:2015:1A, 1B, 1C, 1D, 1E & 1F (continued)
Part UnitsMurray Valley Encephalitis Virus - Units & Cut Off Values
Neg Equ Pos
40002 Ct 40 37-39 3660003 Ct 0 39-40 1-3980069 - No specific band seen - Specific band seen
Part UnitsWest Nile Virus - Units & Cut Off Values
Neg Equ Pos
40002 Ct 40 37-39 3660003 Ct 0 39-40 1-3980069 - No specific band seen - Specific band seen89082 Ct 45 40 2989112 No units available
Part UnitsYellow Fever Virus - Units & Cut Off Values
Neg Equ Pos
40002 Ct 40 37-39 3680069 - No specific band seen - Specific band seen
80071 Cp No product formation at meltingcurve analysis
Product formation at meltingcurve analysis
89082 Ct 45 40 2989112 No units available
© 2015 RCPA Quality Assurance Programs Pty Ltd. All rights reservedReport Issued Monday, 18 May 2015Page Page 23 of 28
Flavivirus Extraction Information for Survey MAF1:2015:1A, 1B, 1C, 1D, 1E & 1F
PartFlavivirus Group - Extraction
Manufacturer Kit Name Lot No. Expiry Date Extraction Vol Elution Vol
10003 Qiagen QIAamp Viral RNA Mini Kit 148026903 Apr 16 140 6040002 Qiagen EZ1 Virus Mini Kit v2.0 148038536 Mar 16 200 9080071 Qiagen QIAamp Viral RNA Mini Kit 148052920 Oct 16 140 6083000 Qiagen QIAamp Viral RNA Mini Kit 148017918 Jan 16 140 5089112 Qiagen QIAamp Viral RNA Mini Kit 148013001 Nov 15 140 60
Part
Dengue Virus - Extraction
Manufacturer Kit Name Automation Lot No. ExpiryDate
ExtractionVol
ElutionVol
1187 Qiagen QIAamp Viral RNA Mini Kit - 148035479 May 16 140 6010003 Qiagen QIAamp Viral RNA Mini Kit - 148026903 Apr 16 140 6040002 Qiagen EZ1 Virus Mini Kit v2.0 - 148038536 Mar 16 200 9060003 Qiagen QIAamp Viral RNA Mini Kit - 148044563 Aug 16 160 10080011 bioMerieux NucliSens EasyMag Z015NA Dec 15 200 2580042 Qiagen EZ1 Virus Mini Kit v2.0 - 148046999 Nov 15 200 6080052 Qiagen EZ1 Virus Mini Kit v2.0 EZ1 BioRobot 148046732 Nov 15 400 6080069 Qiagen QIAamp Viral RNA Mini Kit - 148045591 Sep 16 140 6080071 Qiagen QIAamp Viral RNA Mini Kit - 148052920 Oct 16 140 6083000 Qiagen QIAamp Viral RNA Mini Kit - 148017918 Jan 16 140 50
89082 RocheDiagnostics
MagNA Pure Compact Nucleic AcidIsolation Kit I MagNA Pure 10771500 Dec 15 200 50
89112 Qiagen QIAamp Viral RNA Mini Kit - 148013001 Nov 15 140 60
Part
Japanese Encephalitis Virus - Extraction
Manufacturer Kit Name Automation Lot No. ExpiryDate
ExtractionVol
ElutionVol
10003 Qiagen QIAamp Viral RNA Mini Kit - 148026903 Apr 16 140 6040002 Qiagen EZ1 Virus Mini Kit v2.0 - 148038536 Mar 16 200 9060003 Qiagen QIAamp Viral RNA Mini Kit - 148044563 Aug 16 160 10080069 Qiagen QIAamp Viral RNA Mini Kit - 148045591 Sep 16 140 6080071 Qiagen QIAamp Viral RNA Mini Kit - 148052920 Oct 16 140 6083000 Qiagen QIAamp Viral RNA Mini Kit - 148017918 Jan 16 140 50
89082 RocheDiagnostics
MagNA Pure Compact Nucleic AcidIsolation Kit I MagNA Pure 10771500 Dec 15 200 50
PartKunjin Virus - Extraction
Manufacturer Kit Name Lot No. Expiry Date Extraction Vol Elution Vol
40002 Qiagen EZ1 Virus Mini Kit v2.0 148038536 Mar 16 200 9060003 Qiagen QIAamp Viral RNA Mini Kit 148044563 Aug 16 160 100
© 2015 RCPA Quality Assurance Programs Pty Ltd. All rights reservedReport Issued Monday, 18 May 2015Page Page 24 of 28
Flavivirus Extraction Information for Survey MAF1:2015:1A, 1B, 1C, 1D, 1E & 1F (continued)
PartMurray Valley Encephalitis Virus - Extraction
Manufacturer Kit Name Lot No. Expiry Date Extraction Vol Elution Vol
40002 Qiagen EZ1 Virus Mini Kit v2.0 148038536 Mar 16 200 9060003 Qiagen QIAamp Viral RNA Mini Kit 148044563 Aug 16 160 10080069 Qiagen QIAamp Viral RNA Mini Kit 148045591 Sep 16 140 60
Part
West Nile Virus - Extraction
Manufacturer Kit Name Automation Lot No. Expiry Date ExtractionVol
ElutionVol
40002 Qiagen EZ1 Virus Mini Kit v2.0 - 148038536 Mar 16 200 9060003 Qiagen QIAamp Viral RNA Mini Kit - 148044563 Aug 16 160 10080069 Qiagen QIAamp Viral RNA Mini Kit - 148045591 Sep 16 140 60
89082 RocheDiagnostics
MagNA Pure Compact Nucleic AcidIsolation Kit I MagNA Pure 10771500 Dec 15 200 50
89112 Qiagen QIAamp Viral RNA Mini Kit - 148013001 Nov 15 140 60
Part
Yellow Fever Virus - Extraction
Manufacturer Kit Name Automation Lot No. Expiry Date ExtractionVol
ElutionVol
40002 Qiagen EZ1 Virus Mini Kit v2.0 - 148038536 Mar 16 200 9080069 Qiagen QIAamp Viral RNA Mini Kit - 148045591 Sep 16 140 6080071 Qiagen QIAamp Viral RNA Mini Kit - 148052920 Oct 16 140 60
89082 RocheDiagnostics
MagNA Pure Compact Nucleic AcidIsolation Kit I MagNA Pure 10771500 Dec 15 200 50
89112 Qiagen QIAamp Viral RNA Mini Kit - 148013001 Nov 15 140 60
© 2015 RCPA Quality Assurance Programs Pty Ltd. All rights reservedReport Issued Monday, 18 May 2015Page Page 25 of 28
Flavivirus Amplification Information for Survey MAF1:2015:1A, 1B, 1C, 1D, 1E & 1F
Part
Flavivirus Group - Amplification
Manufacturer Kit Name Method Automation Lot No. ExpiryDate
MasterVol
SampleVol
Single/Nested
10003 In house - RT PCR Eppendorf GradientS Thermalcycler NA NA 37 3 Nested
40002 In house - RT PCR Eppendorf GradientS Thermalcycler 11022015 Feb 16 15 5 Nested
80071 Qiagen QuantiFastSybrGreen RT-PCR LightCycler 2.0 148034258 Sep 15 20 5 Single round
83000 Bioline UK MyTaq One-StepRT-PCR - ABI Veriti MTOS10-
112C May 15 20 5 Nested
89112 Roche AppliedScience
Titan One tubeRT-PCR RT-PCR AB 2720 Thermal
Cycler 10774220 Oct 15 40 10 Single round
Part
Dengue Virus - Amplification
Manufacturer Kit Name Method Automation Lot No. ExpiryDate
MasterVol
SampleVol
Single/Nested
1187 AIT BiotechPte Ltd
abTes TM DEN 4qPCR 1 Kit Real Time PCR Biorad CFX96 C155-26 Aug 15 20 5 Single round
10003 In house - TaqMan RT-PCR ABI Prism NA NA 17 3 Single round40002 In house - TaqMan RT-PCR Rotorgene 18022015 Feb 16 15 5 Single round60003 In house - TaqMan RT-PCR Rotorgene NA NA 12 8 Single round
80011 Invitrogen SuperScript IIIqRT-PCR RT PCR - 1565698 Apr 16 17 3 Single round
80042 In house - Real-time PCR ABI 7500 N/A NA 15 5 Single round
80052 Invitrogen SuperScript IIIqRT-PCR RT PCR - 1507789 Jan 16 20 10 Single round
80069 AITbiotech abTES DEN 5qPCR RT-PCR CFX96 C151-24 May 15 20 5 Single round
80071 RocheDiagnostics
LightCycler RNAAmplification kitHypro
RT-PCR Light Cycler 10772000 Aug 16 9 1 Single round
83000 BioRad iScript One-StepRT-PCR iQ5 172-5141 Jul 16 20 5 Single round
89082 In house - TaqMan RT-PCR LightCycler 110315 Mar 16 15 5 Single round
89112 AltonaDiagnostics
RealStar DengueRT PCR kit Real-time PCR Light cycler
Roche 96 2810-K-1410 Jan 15 25 25 Single round
Part
Japanese Encephalitis Virus - Amplification
Manufacturer Kit Name Method Automation Lot No. ExpiryDate
MasterVol
SampleVol
Single/Nested
10003 In house - TaqMan RT-PCR ABI Prism NA NA 17 3 Single round40002 In house - TaqMan RT-PCR Rotorgene 11022015 Feb 16 15 5 Single round60003 In house - TaqMan RT-PCR Rotorgene NA NA Single round
80069 Qiagen OneStep RT-PCR Kit RT-PCR - 148045112 Sep 15 20 5 Nested
80071 In house - Real-time Light Cycler 10035000 Sep 15 15 5 Single round
83000 Qiagen OneStep RT-PCR Kit RT-PCR - 148030425 Jul 15 20 5 Single round
89082 In house - TaqMan RT-PCR LightCycler 030215 Feb 16 15 5 Single round
© 2015 RCPA Quality Assurance Programs Pty Ltd. All rights reservedReport Issued Monday, 18 May 2015Page Page 26 of 28
Flavivirus Amplification Information for Survey MAF1:2015:1A, 1B, 1C, 1D, 1E & 1F (continued)
Part
Kunjin Virus - Amplification
Manufacturer Kit Name Method Automation Lot No. ExpiryDate
MasterVol
SampleVol
Single/Nested
40002 In house - TaqMan RT-PCR ABI Prism 29012015 Jan 16 15 5 Single round60003 In house - TaqMan RT-PCR Rotorgene NA NA 12 8 Single round
Part
Murray Valley Encephalitis Virus - Amplification
Manufacturer Kit Name Method Automation Lot No. ExpiryDate
MasterVol
SampleVol
Single/Nested
40002 In house - TaqMan RT-PCR Rotorgene 07012015 Jan 16 15 5 Single round60003 In house - TaqMan RT-PCR Rotorgene NA NA 12 8 Single round
80069 Qiagen OneStep RT-PCR Kit RT-PCR - 148045112 Sep 15 20 5 Single round
Part
West Nile Virus - Amplification
Manufacturer Kit Name Method Automation Lot No. ExpiryDate
MasterVol
SampleVol
Single/Nested
40002 In house - TaqMan RT-PCR Rotorgene 05022015 Feb 16 15 5 Single round60003 In house - TaqMan RT-PCR Rotorgene NA NA Single round
80069 Qiagen OneStep RT-PCR Kit RT-PCR - 148045112 Sep 15 20 5 Single round
89082 In house - TaqMan RT-PCR LightCycler 300414 Apr 15 15 5 Single round
89112 Roche AppliedScience
Titan Onetube RT-PCR RT-PCR AB 2720
Thermal Cycler 10774220 Oct 15 40 10 Single round
Part
Yellow Fever Virus - Amplification
Manufacturer Kit Name Method Automation Lot No. ExpiryDate
MasterVol
SampleVol
Single/Nested
40002 In house - TaqMan RT-PCR Rotorgene 10122014 Dec 15 15 5 Single round
80069 Qiagen OneStep RT-PCR Kit RT-PCR - 148045112 Sep 15 20 5 Single round
80071 In house - Real-time PCR Light Cycler 10035000 Sep 15 9 1 Single round89082 In house - TaqMan RT-PCR LightCycler 110315 Mar 16 15 5 Single round
89112 Roche AppliedScience
Titan Onetube RT-PCR RT-PCR AB 2720
Thermal Cycler 10774220 Oct 15 40 10 Single round
© 2015 RCPA Quality Assurance Programs Pty Ltd. All rights reservedReport Issued Monday, 18 May 2015Page Page 27 of 28
Flavivirus Detection Information for Survey MAF1:2015:1A, 1B, 1C, 1D, 1E & 1F
PartFlavivirus Group - Detection
Manufacturer Kit Name Method Automation Lot No. Expiry Date Single/Nested
10003 In house - Gel electrophoresis - NA NA Nested
40002 Qiagen QIAxcel DNA HighResolution Kit QIAxcel C131031033 May 15 Nested
80071 Qiagen QuantiFastSybrGreen RT-PCR LightCycler 2.0 148034258 Sep 15 Single round
83000 In house - Gel electrophoresis - 0000398806 Nov 15 Single round
89112 UV Products UV Transilluminator GeneSnapfrom SynGene IN HOUSE Oct 15 Single round
Part
Dengue Virus - Detection
Manufacturer Kit Name Method Automation Lot No. ExpiryDate Single/Nested
1187 AIT BiotechPte Ltd
abTes TM DEN 4qPCR 1 Kit Real Time PCR Biorad CFX96 C155-26 Aug 15 Single round
10003 In house - TaqMan RT-PCR ABI Prism NA NA Single round40002 In house - TaqMan RT-PCR Rotorgene 18022015 Feb 16 Single round60003 In house - TaqMan RT-PCR Rotorgene NA NA Nested80011 In house - TaqMan RT-PCR LightCycler 1565698 Apr 16 Single round80042 In house - Real-time PCR ABI 7500 N/A NA Single round80052 In house - TaqMan RT-PCR Rotorgene 6089093 NA Single round80069 AITbiotech abTES DEN 5 qPCR RT-PCR CFX96 C151-24 May 15 Single round80071 In house - Real-time PCR Light Cycler 10772000 Aug 16 Single round
83000 BioRad iScript One-Step RT-PCR iQ5 172-5141 Jul 16 Single round
89082 In house - TaqMan RT-PCR LightCycler 110315 Mar 16 Single round
89112 AltonaDiagnostics
RealStar Dengue RT-PCR kit Real-time PCR Light Cycler
Roche 96 2810-K-1410 Jan 15 Single round
Part
Japanese Encephalitis Virus - Detection
Manufacturer Kit Name Method Automation Lot No. ExpiryDate Single/Nested
10003 In house - TaqMan RT-PCR ABI Prism NA NA Single round40002 In house - TaqMan RT-PCR Rotorgene 11022015 Feb 16 Single round60003 In house - TaqMan RT-PCR Rotorgene NA NA Single round80069 In house - Gel electrophoresis - - NA80071 In house - Real-time Light Cycler 10035000 Sep 15 Single round
83000 Qiagen Quantitect ProbeRT-PCR kit
Fluorescent (FAMchannel) BioRad CFX 96 148030425 Jul 15 Single round
89082 In house - TaqMan RT-PCR LightCycler 030215 Feb 16 Single round
© 2015 RCPA Quality Assurance Programs Pty Ltd. All rights reservedReport Issued Monday, 18 May 2015Page Page 28 of 28
Flavivirus Detection Information for Survey MAF1:2015:1A, 1B, 1C, 1D, 1E & 1F (continued)
Part
Kunjin Virus - Detection
Manufacturer Kit Name Method Automation Lot No. ExpiryDate Single/Nested
40002 In house - TaqMan RT-PCR Rotorgene 29012015 Jan 16 Single round60003 In house - TaqMan RT-PCR Rotorgene NA NA Single round
Part
Murray Valley Encephalitis Virus - Detection
Manufacturer Kit Name Method Automation Lot No. ExpiryDate Single/Nested
40002 In house - TaqMan RT-PCR Rotorgene 07012015 Jan 16 Single round60003 In house - TaqMan RT-PCR Rotorgene NA NA Single round80069 In house - Gel electrophoresis - NA NA
Part
West Nile Virus - Detection
Manufacturer Kit Name Method Automation Lot No. ExpiryDate Single/Nested
40002 In house - TaqMan RT-PCR Rotorgene 05022015 Feb 16 Single round60003 In house - TaqMan RT-PCR Rotorgene NA NA Single round80069 In house - Gel electrophoresis - NA NA89082 In house - TaqMan RT-PCR LightCycler 300414 Apr 15 Single round
89112 UV Products UVTransilluminator
GeneSnapfrom SynGene IN HOUSE Oct 15 Single round
Part
Yellow Fever Virus - Detection
Manufacturer Kit Name Method Automation Lot No. ExpiryDate Single/Nested
40002 In house - TaqMan RT-PCR Rotorgene 10122014 Dec 15 Single round80069 In house - Gel electrophoresis - NA NA80071 In house - Real-time PCR Light Cycler 10035000 Sep 15 Single round89082 In house - TaqMan RT-PCR LightCycler 110315 Mar 16 Single round
89112 UV Products UVTransilluminator
GeneSnapfrom SynGene IN HOUSE Oct 15 Single round