5 th <strong>European</strong> <strong>Congress</strong> <strong>of</strong> <strong>Virology</strong>24. 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 <strong>Virology</strong> 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 (<strong>European</strong>,African 1 and 2, Asian, North American and Asian American). In ourstudy, 50% <strong>of</strong> CIN2+ and 24% <strong>of</strong> CIN2 cases had HPV 16. Our aim wasto find out if some variants were more present in our region and if some <strong>of</strong>them 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% <strong>of</strong> them have been sequenced successfully,and second, samples below 3 log, only 15.2% were sequenced (n=46).Most <strong>of</strong> the samples sequenced presented an <strong>European</strong> 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 <strong>European</strong> 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 <strong>of</strong> 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 vi<strong>rologie</strong>, Hôpital Militaire d’instru, Rabat,MOROCCO; 2 Laboratoire de Vi<strong>rologie</strong>, CNRS UMR5234, WHO Accredited(HIV Resistance), Universite Victor Segalen, Bordeaux, FRANCEDetermination <strong>of</strong> human immunodeficiency virus tropism plays a crucialrole in AIDS progression and is necessary prior to the use <strong>of</strong> CCR5 antagonists.The aim <strong>of</strong> this study is to investigate predicted HIV 1 co receptorusage in HIV 1isolates in Morocco. Methods: A total <strong>of</strong> 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 <strong>of</strong> V3 region <strong>of</strong>gp120 env gene. The obtained fragments were sequenced on both strands.HIV 1 subtype analysis was determined by phylogenetic analysis <strong>of</strong> RTand gp120 sequences. The genotypic interpretation <strong>of</strong> HIV 1 tropism wasperformed using the Geno2phenocoreceptor approach. The Geno2phenointerpretation was made with the false positive rates (FPRs) for prediction<strong>of</strong> X4 variants FPRs <strong>of</strong> 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% <strong>of</strong> 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% <strong>of</strong> subtypeB and 90% <strong>of</strong> CRF02 AG are also R5, indicating a higher prevalence <strong>of</strong>R5 (89%) than X4 (2%) or R5X4 (9%). The co receptor usage <strong>of</strong> HIV 1seems to be not associated with the different subtypes. Conclusion: Thehigher prevalence <strong>of</strong> R5 suggest that CCR5 antagonists will be promisingdrugs for future AIDS treatment in Morocco.REF 494Recent Emergence and Spread <strong>of</strong> a Phylogenetic Lineage <strong>of</strong> 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 <strong>of</strong> Microbiology, Tribhuvan University,Kathmandu, NEPALRabies is a zoonotic disease that is endemic in many parts <strong>of</strong> the developingworld, especially in Africa and in Asia. However its epidemiologyremains largely unappreciated in much <strong>of</strong> these regions, such as in Nepal,where limited information is available about the spatiotemporal dynamics<strong>of</strong> the main etiological agent, the rabies virus (RABV). In this study,we describe for the first time the phylogenetic diversity and evolution <strong>of</strong>RABV circulating in Nepal, as well as their geographical relationshipswithin the broader region. A total <strong>of</strong> 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 <strong>of</strong> a surprising wide genetic diversity <strong>of</strong>RABV, with the co existence <strong>of</strong> 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 <strong>of</strong> rabies introduction from neighbouring countries. In addition,specific phylogenetic and temporal evolution analysis <strong>of</strong> 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.Vi<strong>rologie</strong>, Vol 17, supplément 2, septembre 2013S257
5 th <strong>European</strong> <strong>Congress</strong> <strong>of</strong> <strong>Virology</strong>REF 495Cytomegalovirus gB, gpUL144 and pUS28 genotypes distributionamong Polish childrenMiroslawa STUDZINSKA 1 , Edyta PARADOWSKA 1 , MiroslawaSTUDZINSKA 1 , Patrycja SUSKI 1 , Agnieszka JABLONSKA 1 ,Katarzyna DZIERZANOWSKA FANGRAT 2 , BeataKASZTELEWICZ 2 , Malgorzata WISNIEWSKA LIGIER 3 , TeresaWOZNIAKOWSKA GESICKA 3 , Barbara ZAWILINSKA 4 , Zbigniew J.LESNIKOWSKI 11 Laboratory <strong>of</strong> Molecular <strong>Virology</strong> and Biological Chemistry, Institute<strong>of</strong> Medical Biology Polish Academy <strong>of</strong> Sciences, Lodz, POLAND;2 Department <strong>of</strong> Microbiology and Clinical Immunology, Children MemorialHealth Institute, Warsaw, POLAND; 3 III Department <strong>of</strong> Peadiatrics,Research Institute Polish Mother’s Memorial Hospital, Lodz, POLAND;4 Department <strong>of</strong> <strong>Virology</strong>, Jagiellonian University Medical College, Cracow,POLANDHuman cytomegalovirus (HCMV) is the most common cause <strong>of</strong> viralintrauterine infection, affecting 0.5 2.0% <strong>of</strong> all live births. The association<strong>of</strong> genetic polymorphisms in some particular genes with the occurrence <strong>of</strong>congenital infection is debated. The aim <strong>of</strong> our studies was to determinethe distribution <strong>of</strong> the HCMV genotypes isolated from Polish children andto recognize the relationship between genotype, viral load, and clinicalsequelaes. We analyzed the gB, gpUL144 and pUS28 genetic variations incongenitally HCMV infected newborns (n=34) and infants with postnatalor unproven congenital HCMV infection (n=90). Genotyping was performedby sequence analysis <strong>of</strong> PCR amplified fragments, and viral load wasmeasured by quantitative real time PCR. Total genomic DNA was extractedfrom blood, plasma, peripheral blood mononuclear cells (PBMC) and urinesamples. Our results demonstrated that HCMV genotypes gB1and A1 wereprevalent in congenitally infected newborns (p
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