rologie i - European Congress of Virology

5 th European Congress of Virology24. TYPING OF VIRUSES (MOLECULAREPIDEMIOLOGY, TAXONOMY)Posters: REF 492 to REF 507REF 492HPV 16 variants isolated from general population in finistere, WestBrittany, FranceChristopher PAYAN 1 , Aureliane MICHAUD 1 , Stephanie GOURIOU 1 ,Sylvain ROSEC 2 , Sophie VALLET 1 , Adissa TRAN 1 , Yvon FOLL 3 ,Francoise BOMMELAERT 3 , Francoise CHARLES 4 , Edith POSTEC 5 ,Michel COLLET 51 Virology Lubem Chru Ufr Medecine, Brest, FRANCE; 2 Cic Inserm0502, Brest, FRANCE; 3 Adec29, Brest, FRANCE; 4 Cytologie Chru, Brest,FRANCE; 5 Gynecology Chru, Brest, FRANCEObjectives: From a HPV DNA test in urine (PapU29 study; Payan, 2007),we typed all HPV 16 positive samples to determine variants (European,African 1 and 2, Asian, North American and Asian American). In ourstudy, 50% of CIN2+ and 24% of CIN2 cases had HPV 16. Our aim wasto find out if some variants were more present in our region and if some ofthem were more virulent than others. Methods: Urine samples were obtainedfrom 25 65 years old women (n=3115) and HPV 16 positive (n=88),we performed PCR with SiHa HPV 16+ cells as control. Since the concentrationrange was fluctuent (from 1 to 7 log copies per mL), several PCRin the E6 gene and adjustments were necessary. We established that threePCR were needed including two nested. They were followed by electrophoresisand, when positive, DNA samples were sequenced and analyzedusing bioinformatic tools (Staaden Package and BioEdit). Results: Twogroups have been characterized. First, samples with more than 3 log DNAcopies per mL (n=42), 83% of them have been sequenced successfully,and second, samples below 3 log, only 15.2% were sequenced (n=46).Most of the samples sequenced presented an European variant, 57% werethe mutant 350G and with some minor differencies; only two were an Af2 variant located in the same city.Conclusion: It does not appear that there is a link between variant andCIN2+ cases, we demonstrated that the European variant is more presentthan any other in West Brittany. Nevertheless, more HPV 16 patients shouldbe done to confirm those results. Grants from the French Ligue contre leCancer.REF 493Genotypic prediction of HIV 1 isolates tropism in MoroccoHicham EL ANNAZ 1 , Patricia RECORDON PINSON 2 , RidaTAGAJDID 1 , Bouchra BELEFQUIH 1 , Safae EL KOCHRI 1 , RachidABBI 1 , Siham OUMAKHIR 1 , Herve FLEURY 2 , Saad MRANI 11 Université Mohamed V Souissi, Faculté de Médecine et de Pharmaciede Rabat. Laboratoire de virologie, Hôpital Militaire d’instru, Rabat,MOROCCO; 2 Laboratoire de Virologie, CNRS UMR5234, WHO Accredited(HIV Resistance), Universite Victor Segalen, Bordeaux, FRANCEDetermination of human immunodeficiency virus tropism plays a crucialrole in AIDS progression and is necessary prior to the use of CCR5 antagonists.The aim of this study is to investigate predicted HIV 1 co receptorusage in HIV 1isolates in Morocco. Methods: A total of 45 MoroccanHIV 1 infected patients followed up at the Mohammed V Military TeachingHospital in Rabat were included in this study. All patients were drugNaive to treatment. The viral RNA was used for reverse transcription polymerasechain reaction (RT PCR) followed by a nested PCR of V3 region ofgp120 env gene. The obtained fragments were sequenced on both strands.HIV 1 subtype analysis was determined by phylogenetic analysis of RTand gp120 sequences. The genotypic interpretation of HIV 1 tropism wasperformed using the Geno2phenocoreceptor approach. The Geno2phenointerpretation was made with the false positive rates (FPRs) for predictionof X4 variants FPRs of 10% and with the clinical parameters data (the viralload and the CD4 value). Results: Among the 45 HIV 1 infected treatmentnaive patients, 38 (84%) are males. Their mean age is 41 [35 48] years.The WHO clinical classification indicated that 13% of these patients are atstage 1, 25% at stage 2, 30% at stage 3 and 32% at stage 4. The median HIV1 viral load and CD4 cell count HIV 1 infected patients is 45700 [10140151000] copies/ml and 228 [107 419] cells/mm3 respectively. The viralsubtypes are subtype B (71%), CRF02 AG (20%) and C (9%). All subtypeC appear to use CCR5 for cell entry (R5 strains), while 87% of subtypeB and 90% of CRF02 AG are also R5, indicating a higher prevalence ofR5 (89%) than X4 (2%) or R5X4 (9%). The co receptor usage of HIV 1seems to be not associated with the different subtypes. Conclusion: Thehigher prevalence of R5 suggest that CCR5 antagonists will be promisingdrugs for future AIDS treatment in Morocco.REF 494Recent Emergence and Spread of a Phylogenetic Lineage of RabiesVirus in NepalLaurent DACHEUX 2 , Ganesh R. PANT 1 , Rachel LAVENIR 2 , Frank Y.K. WONG 3 , Andrea CERTOMA 3 , Florence LARROUS 2 , Dwij R.BHATTA 4 , Hervé BOURHY 2 , Vittoria STEVENS 31 Rabies Vaccine Production Laboratory, Kathmandu, NEPAL 2 InstitutPasteur, Unit Lyssavirus Dynamics and Host Adaptation, National ReferenceCentre for Rabies, Paris, FRANCE; 3 Australian Animal HealthLaboratory, CSIRO Animal Food and Health Sciences, Geelong, AUS-TRALIA; 4 Central Department of Microbiology, Tribhuvan University,Kathmandu, NEPALRabies is a zoonotic disease that is endemic in many parts of the developingworld, especially in Africa and in Asia. However its epidemiologyremains largely unappreciated in much of these regions, such as in Nepal,where limited information is available about the spatiotemporal dynamicsof the main etiological agent, the rabies virus (RABV). In this study,we describe for the first time the phylogenetic diversity and evolution ofRABV circulating in Nepal, as well as their geographical relationshipswithin the broader region. A total of 24 new isolates obtained from Nepaland collected from 2003 to 2011 were full length sequenced for both thenucleoprotein and the glycoprotein genes, and analysed using neighborjoining and maximum likelihood phylogenetic methods with representativeviruses from all over the world. Despite Nepal’s limited land surfaceand its particular geographical position within the Indian subcontinent,our study revealed the presence of a surprising wide genetic diversity ofRABV, with the co existence of three different phylogenetic groups: anIndian subcontinent clade and two different Arctic like sub clades withinthe Arctic related clade. This observation suggests at least two independentepisodes of rabies introduction from neighbouring countries. In addition,specific phylogenetic and temporal evolution analysis of viruses within theArctic related clade has identified a new recently emerged RABV lineagewe named as the Arctic like 3 (AL 3) sub clade that is already widelyspread in Nepal.Virologie, Vol 17, supplément 2, septembre 2013S257