rologie i - European Congress of Virology

5 th European Congress of Virologyness cases were sent weekly by sentinel GPs to the two Greek NationalInfluenza Centres. Specimens were also received from outpatients and hospitalisedcases. RNA was extracted from 36 influenza A(H3N2) strains. RTPCR and sequencing was performed and nucleotide sequences were usedfor the molecular and phylogenetic analysis using the neighbour joining(NJ) method implemented in MEGA 5.0. Results: phylogenetic analysisof Greek H3N2 strains indicated clustering within the Victoria/208 cladei.e. groups 3A, 3B, 3C and 6. Interestingly no isolates fell into the geneticPerth/16 clade characterized by the H3N2 component of the 2011 12influenza vaccine. Variations on antigenic sites accumulated on the A, E,C and D, while B antigenic sites were conserved. Amino acid analysis ofA/Athens/16/2012 strain revealed the emergence of 4 amino acid substitutionsin 2 different antigenic sites indicating potential antigenic drift. All ofthe Greek A(H3N2) examined strains possessed the L183H substitutionthat affected the HA receptor binding site. Variations were also observedon the N linked glycosylation sites of HA. All 2011 12 isolates hadan altered potential N linked glycosylation site at amino acids 144–146.Four Southern greek stains that belonged to the 3C phylogenetic group,also possessed an additional glycosylation site, due to substitution S45N.These alterations may have influenced the function of the glycoprotein ofthose viruses.Conclusion: Our findings confirmed the genetic instability of influenzatype A (H3N2) viruses and the importance of continuous molecular surveillancefor the effective management of viral epidemics.REF 378Molecular characterisation of Human Hepatitis E virus from ItalyGiuseppina LA ROSA 1 , Marta FRATINI 1 , Marcello IACONELLI 1 ,Michele MUSCILLO 1 , Simonetta DELLA LIBERA 1 , MicheleEQUESTRE 2 , Angela CANDIDO 2 , Stefania TAFFON 2 , PaolaCHIONNE 2 , Elisabetta MADONNA 2 , Stefano DETTORI 2 , RobertoGIUSEPPETTI 2 , Roberto BRUNI 2 , Anna Rita CICCAGLIONE 21 Department of Environment and Primary Prevention, Istituto Superioredi Sanità, Rome, ITALY; 2 Department of Infectious, Parasitic and ImmuneMediated Diseases, Istituto Superiore di Sanità, Rome, ITALYHuman hepatitis E virus (HEV) is an emerging pathogen in industrialisedcountries. We re analysed a collection of serum samples previouslyconfirmed as HEV positive by anti HEV IgM and IgG assays as well as byReal Time PCR, with the aim of shedding light on the molecular epidemiologyof HEV in Italy. Samples were analyzed by published and newlydesigned PCR assays, using four sets of primers targeting the ORF1 andORF2, followed by phylogenetic analysis. Despite the use of four differentassays, not all samples tested positive by nested PCR. Moreover,no single method was able to detect all positive samples. Most sequencesgrouped with genotype 1, and originated from patients who had travelledto endemic areas, while the minority belonged to genotype 3, and originatedfrom Italian patients with no travel history. Phylogenetic analysisshowed a match between sequences derived from patients with travel relatedHEV and sequences from the geographical regions in which infectionwas acquired (mostly Bangladesh and India). Sequences from patientswith no travel history clustered on the same branch with published swineHEV isolates. Although autochthonous cases do occur, this study confirmsthat HEV in Italy is predominantly travel related. Broad range methodsfor molecular diagnosis of HEV are still in need of improvement. Thestudy was co funded by the Italian National Centre for the Prevention andControl of Diseases (CCM) project Hepatitis E Surveillance in Italy: anemerging disease in industrialised countries, and by the EU 7th Frameworkprogramme PREDEMICS project.REF 379Human Hepatitis E virus in urban wastewaters in ItalyGiuseppina LA ROSA, Marta FRATINI, Marcello IACONELLI,Simonetta DELLA LIBERA, Michele MUSCILLODepartment of Environment and Primary Prevention, Istituto Superiore diSanità, Rome, ITALYIn industrialised countries, Hepatitis E infection (HEV) is usuallyassociated with travel to endemic areas, but a growing number of sporadiccases are also seen in patients with no travel history. The objective ofthe present study was to investigate the occurrence of HEV in Italythrough the molecular screening of raw sewage samples collected fromurban wastewater treatment plants (WTPs). Three sets of broad rangePCR primers targeting the ORF1 (MTase and RdRp) and ORF2 (capsid)were used to examine 200 sewage samples collected throughout Italy(38 WTPs). PCR products were then subjected to sequencing andphylogenetic analysis. Despite the use of different assays, HEV RNAwas detected only in two samples, both belonging to genotype 3. On thephylogenetic tree, sequences clustered on the same branch with HEVisolates identified in serum samples of Italian patients with autochthonousHEV (no travel history). Although different studies indicate that HEVin Italy is predominantly travel related, in this study no genotype 1was detected. The study was co funded by the Italian National Centrefor the Prevention and Control of Diseases (CCM) project Hepatitis ESurveillance in Italy: an emerging disease in industrialised countries, andby the EU 7th Framework programme PREDEMICS project.REF 380High seroprevalence of Hepatits E virus among Egyptian BloodDonorsLobna METWALLY 2 , Endale TADESSE 1 , Alaa El Din SAAD ABD ELHAMID 31 Department of Medical Laboratory Science, College of Medicineand Health Sciences, Hawassa University, Hawassa, ETHIOPIA;2 Department of Microbiology, Faculty of Medicine, Suez Canal University,Ismailia, EGYPT; 3 Department of Clinical Pathology, Faculty of Medicine,Suez Canal University, Ismailia, EGYPTAim: the aim of this study was to assess seroprevalence of hepatitis E virus(HEV) among blood donors attending blood transfusion center of SuezCanal University Hospital during the study period. Method: 488 subjectsconsisted of 137 anti HCV positive donors, 35 HBsAg positive donors, and316 blood donors who were negative HBsAg, anti HCV and HIV attendingSuez Canal University Hospital blood transfusion center were included inthis study. All the study subjects were tested for anti HEV by ELISA andthe positive samples were further tested for HEV RNA by reveres transcriptasepolymerase chain reaction (RT PCR) method. Results: in a total of488 subjects, 102 (20.9%) were positive for anti HEV by ELISA. Anti HEVwas detected in (56/316) 17.7%, (10/35) 28.57%, and (36/137)26.28%, ofblood donors negative for both HBsAg & Anti HCV, HBsAg positive andAnti HCV positive donors, respectively. No significant (P>0.05) associationwas found between anti HEV positivity and HBsAg positivity andanti HCV positivity subjects. Anti HEV among blood donors with normalALT level (40) who were negative for HCV Ab, HBs Ag, and HIVwas 19.6%(43/219) and 13.4%(13/97) respectively. There was no statisticalsignificance (P>0.05) between the increase ALT level and Anti HEV.Conclusion: Seroprevalence of HEV antibody among blood donors in ourstudy in Ismailia, Egypt is high, but we cannot recommend screening of allblood donors for HEV until more data becomes available and until moreis known about the parenteral rout of transmission of HEV.Virologie, Vol 17, supplément 2, septembre 2013S225