HUMAN enterovirus AND VIRAL MENINGITIS IN SOUTHERN BRAZIL

1. HUMAN enterovirus AND VIRAL MENINGITIS IN SOUTHERN BRAZIL LuineRosele Renaud Vidal Universidade Federal do Paraná Financial Support: Secretaria de Ciência e Tecnologia do Estado do Paraná- SETI - PR

2. Research Background • NotifiableDisease in theNational Health System • Around 1000 meningitis cases per year • Alletiology • Estimated 60% - viral etiology

3. Research Background DistributionofViral Meningitisin Curitiba City byNeighborhood

4. Research Background 460 CSF Samples July 2005 – December 2006 13% Positive

5. Research Background Vidal et al, 2011

6. Research Background

7. Research Background

8. Research Background • July 2005 – September 2008 • 1010 CSF samples (immunocompromised / immunocompetent) • 4.55% positive bynested PCR (nPCR) • 50% encephalitis • 40% meningoencephalitis • 2.5% mielitis Central NervousSystem (CNS) InfectionsbyHerpesviridaeFamily

9. Research Background Central Nervous System (CNS) Infections by Herpesviridae Family %

10. Molecular characterizationoFHuman enterovirus • July 2005 – December 2006 • 440 CSF samples – meningitispatients • 49 positive samples - 11%

11. MeningitisPatients InclusionCriteria ExclusionCriteria EncephalitisParenquimaEnvolvement BacterialMeningitis Inadequatesample: collection / transport / storage • Acuteonset: < 4 weeks • Fever: > 38 °C • WBC: > 4 cell/mm3 • LymphocytesPredominance • Normal Glucose (≥ 45 mG/dL) • Lactate: < 3.5 mmol/L

12. MeningitisPatients

13. METHODS Casas et al, Journalof Medical Virology,1999 Obersteet al, Virus Research, 1999

14. Seasonality

15. Nucleotidesequencing EV Typing 12 samples

16. Nucleotidesequencing HumanEchovirus 30

17. Nucleotidesequencing ECHOVIRUS 4

18. Nucleotidesequencing HUMAN ECHOVIRUS

19. Conclusions • The molecular methodologywasefectiveandfastto define theetiologyofmeningitis. • CNS infection for EV wasobserved in thesummer (highertemperatures). • The EV typesfoundwereHumanEchovirus • It wasobserved a predominanceof EV infection in the ages of 4 to 17 years old. • It wasnotpossibletoidentifytheetiologicagent in themajorityofthesamples.

20. Currently • Establishing a flowchart for CSF collecting • Introduction of lactate quantification • DNA/RNA Extraction – commercial reagentes • Real Time PCR Changesand Benefits

21. Investigationand Molecular Epidemiology: • Adenoviruses • Flavivirus • Arenavirus • Patientswithsignsandsymptomsofencephalitisandmeningoencephalitis Future Researches

22. Future Researches Human Parechovirus - HPeV Project CSF samplesstoredfrom2007-2014 Objectivetoestablishtheimpactofthevirus

23. General Hospital Universidade Federal do Paraná

24. Team VirologyLaboratory

25. Curitiba City Bothanic Garden