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HUMAN ENTEROVIRUS AND VIRAL MENINGITIS IN SOUTHERN BRAZIL
Luine Rosele Renaud Vidal
Universidade Federal do Paraná
Financial Support: Secretaria de Ciência e Tecnologia do Estado do Paraná- SETI - PR
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Research Background
• Notifiable Disease in the National Health System
• Around 1000 meningitis cases per year
• All etiology• Estimated 60% - viral etiology
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Research Background
Distribution of Viral Meningitis in Curitiba City by Neighborhood
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Research Background
460 CSF Samples July 2005 – December 200613% Positive
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Research Background
Vidal et al, 2011
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Research Background
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Research Background
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Research Background
• July 2005 – September 2008• 1010 CSF samples (immunocompromised / immunocompetent)
• 4.55% positive by nested PCR (nPCR)
• 50% encephalitis• 40% meningoencephalitis• 2.5% mielitis
Central Nervous
System (CNS) Infections by
Herpesviridae Family
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Research Background
Central Nervous
System (CNS) Infections by
Herpesviridae Family
%
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MOLECULAR CHARACTERIZATION OF HUMAN ENTEROVIRUS
• July 2005 – December 2006• 440 CSF samples – meningitis patients• 49 positive samples - 11%
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Meningitis Patients
Inclusion Criteria
• Acute onset: < 4 weeks• Fever: > 38 °C• WBC: > 4 cell/mm3
• Lymphocytes Predominance• Normal Glucose (≥ 45 mG/dL) • Lactate: < 3.5 mmol/L
Exclusion Criteria
• Encephalitis Parenquima Envolvement
• Bacterial Meningitis• Inadequate sample:
collection / transport / storage
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Meningitis Patients
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METHODS
Casas et al, Journal of Medical Virology,1999Oberste et al, Virus Research, 1999
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Seasonality
Ju
l-05
Au
g-0
5
Se
p-0
5
Oc
t-05
No
v-0
5
De
c-0
5
Ja
n-0
6
Fe
b-0
6
Ma
r-06
Ap
r-06
Ma
y-0
6
Ju
n-0
6
Ju
l-06
Au
g-0
6
Se
p-0
6
Oc
t-06
No
v-0
6
De
c-0
6
0
2
4
6
8
10
12
14
16
0
5
10
15
20
25
Positive Samples Average Temp.
Months
Sam
ples
Ave
rage
T
empe
ratu
re
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Nucleotide sequencing
EV Typing
12 samples
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Nucleotide sequencing
Human Echovirus 30
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Nucleotide sequencing
BR2006.767.4
BR2006.03.4
AB268167.1 echo 4 CHINA
AF081643.1 echo 4 USA
AF233373.1 echo 4 ALEMANHA
73
69
0.05
ECHOVIRUS 4
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Nucleotide sequencing
HUMAN ECHOVIRUS
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Conclusions
• The molecular methodology was efective and fast to define the etiology of meningitis.
• CNS infection for EV was observed in the summer (higher temperatures).
• The EV types found were Human Echovirus• It was observed a predominance of EV infection in the ages of 4 to 17 years old.
• It was not possible to identify the etiologic agent in the majority of the samples.
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Currently
• Establishing a flowchart for CSF collecting
• Introduction of lactate quantification • DNA/RNA Extraction – commercial reagentes
• Real Time PCR
Changes and
Benefits
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• Investigation and Molecular Epidemiology:
•Adenoviruses•Flavivirus•Arenavirus
•Patients with signs and symptoms of encephalitis and meningoencephalitis
Future Researches
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Future Researche
s
Human Parechovirus - HPeV Project- CSF samples stored from 2007-2014- Objective to establish the impact of the virus
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General Hospital Universidade Federal
do Paraná
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Team Virology Laboratory
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Curitiba CityBothanic Garden