rologie i - European Congress of Virology

5 th European Congress of VirologySaturday 14 th September 2013, 14h45 – 16h45WORKSHOP 22: “INNOVATIVE METHODS IN VIRALDAIGNOSIS”Chairpersons: Xavier de LAMBALLERIE (Marseille,FRANCE) & Elisabeth PUCHHAMMER (Vienna, AUSTRIA)AmphitheaterKEYNOTE:Molecular diagnostics in the future: how will it look like and what arethe challenges?Hubert G.M. NIESTERSUniversity Medical Center Groningen, Department of Medical Microbiology,Division of Clinical Virology. Hanzeplein 1, 9713 GZ Groningen,THE NETHERLANDSWe all agree that the introduction of molecular diagnostic technologieshas changed clinical virology tremendously over the last few decades.It has been beneficial for patient care and diagnostics, as well assistedthe laboratories in dealing with the rapid detection of new viruses whichcontinuously seem to appear.The expectations and demands for the laboratories has also changed. Regulationsare demanding that the laboratories should be accredited, in whichprocesses around quality control (external and internal) as well as validationand verification of methods and processes are properly documented.Also, the availability of rapid molecular testing of relevant targets and therapid characterization of viruses (type, strain) by sequence analysis, doesbecome more accessible. A short turn-around-time with a lot of clinicallyimportant information does become a trend; not only the quantitative detectionof the virus, but also its sequence characterization. Important in thisturmoil of information will be the communication possibilities between thedifferent technologies within the laboratory (whether this is quality controlrelated or diagnostic result related) as well as between the laboratory andthe clinicians and/or patients.An overview of the new and existing technologies will be presented togetherwith the challenges we still have to overcome.ORAL COMMUNICATIONSResults: Target antigens were selected for the genus flavivirus, and spottedonto nitrocellulose pads on a microarray using a non contact array spotter.Serum samples (N=60) from patients with virological and serologicalconfirmed arboviral infections and 40 control sera of non exposed individualswere incubated in serial 2 fold dilutions followed by incubation withan IgG or IgM specific Cy5 labeled conjugate. After quantifying signalsusing a scanarray scanner, data were analyzed in ‘R’.Profiling of IgG antibodies from the patients confirmed the diagnosis for92% of cases (95% dengue, 82% west Nile, 100% Japanese encephalitisand 100% Usutu virus). Four patients showed reactivities suggesting(co)infections with another virus within the same genus. Specificity wasbetween 92% (dengue) and 100% (West Nile, Japanese encephalitis andUsutu virus) for IgG, and 100% for IgM. Profiling of antibodies in patientsexposed to flaviviruses showed highly discriminatory patterns of reactivityin most patient.REF O170Current mumps outbreak in Belgium: is laboratory confirmation byIgM useful?Liesbeth SEAUX, Elizaveta PADALKOGhent University Hospital, Ghent, BELGIUMNonetheless the vaccination state of the Belgian population is high, we arecurrently facing an outbreak of mumps. This current outbreak started in2012 among Ghent university students. The genotype G variant, not endemicto our regions, was the culprit. Fully immunised patients were infectedbecause current vaccines only contain mumps genotype A antigens as wellas currently available laboratory tests. The aim of this study was to evaluatethe influence of currently circulating genotype G on the performanceof routine mumps IgM test (Enzygnost ® Anti Parotis Virus/IgM, BehringElisa Processor III, Siemens) at the Ghent University Hospital.Based on laboratory information system query for 2012, only patients witha serological request form consistent with the possible clinical pictureof mumps were included. Secondly, the electronic patient records of allwithhold cases were carefully examined.From 234 analysed cases by now, 47 presented with a suspicious clinicalpicture for mumps. Thirty two patients (68%) were between 14 and 28years old. In 35 out of 47 patients (74%) mumps was actually clinicallydiagnosed. Only 13 of these 35 clinically clear cases (37%) showed positivemumps IgM. In total 16 of 47 cases (34%) tested mumps IgM positive.In conclusion, in the current outbreak the routine IgM mumps test was notfitted to confirm the clinical diagnosis. Our study emphasizes that mumpsdiagnosis should be made on clinical basis.REF O169Validation of a flavivirus protein microarray for simultaneous detectionof and differentiation between west Nile, Japanese encephalitis,Usutu and dengue virus immunoglobin G and M antibodiesNatalie CLETON 1,2 , Chantal REUSKEN 1 , Gert Jan GODEKE 1 , JohanREIMERINK 1 , Marion KOOPMANS 1,21 National Institute for Public Health and the Environment, Bilthoven, THENETHERLANDS; 2 Erasmus Medical Center, Rotterdam, THE NETHER-LANDSBackground: The flavivirus genus holds the world’s most widespreadarboviral travel related diseases, causing meningoencephomyelitis, rash,althralgia and hemorrhagic fever. Diagnosis is based mostly on serology,as viremia is short lived. A diagnostic problem is that clinical syndromescaused by different flaviviruses and their geographical distribution overlap,and antibodies cross react extensively in common serological tests.Objective: To assess the potential added value for diagnosis and surveillanceof protein microarray based profiling of antibodies to 4flaviviruses.REF O171Human Papillomavirus Genotyping: Comparison of the seegene AnyplexII HPV28 with the PGMY CHUV AssayRoland SAHLI 1,2 , Christine ESTRADE 1,21 Institute of Microbiology, Centre Hospitalier Universitaire Vaudois andUniversity of Lausanne, Lausanne, SWITZERLAND; 2 WHO HPV LabNetRegional Reference Laboratory for Europe, Lausanne, SWITZERLANDGenotyping of human papillomaviruses (HPV) is essential for surveillanceof HPV vaccines and cervical cancer screening. The PGMY CHUV (PG)assay from the WHO Laboratory Manual allows genotyping of 31 HPV byPCR and reverse blotting hybridization. The Seegene Anyplex II HPV28(H28) is a new real time PCR assay for genotyping and semi quantifying28 HPV. H28 was compared with PG as a reference restricted on their26 common HPV, either retrospectively (309 DNA) or prospectively (84samples) to also evaluate H28 automation (NIMBUS IVD).H28 and PG were fully concordant on 246 (79.6%; 27 neg, 219 pos) out ofthe 309 retrospective DNA. The other 63 (20.4%) cases were either fullyVirologie, Vol 17, supplément 2, septembre 2013S115