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PCR-based next-generation West Nile virus sequencing protocols
Institution:1. Université Tunis El Manar, Institut de la Recherche Vétérinaire de Tunisie, Tunisie;2. Istituto Zooprofilattico Sperimentale dell''Abruzzo e del Molise ‘G. Caporale’, Teramo, Italy;3. Ecole Nationale de Médecine Vétérinaire de Sidi Thabet, Université la Manouba, IRESA, Tunisie;4. OTD centre avicole de Mateur Bizerte Tunisie, Tunisie;1. ErasmusMC, Department of Viroscience, WHO Collaborating Centre for Arbovirus and Viral Hemorrhagic Fever Reference and Research, Rotterdam, the Netherlands;2. Veterinary Pathology Centre, University of Utrecht, the Netherlands;3. GD Animal Health, Deventer, the Netherlands;1. Institute of Novel and Emerging Infectious Diseases, Friedrich-Loeffler-Institut, Greifswald, Insel Riems, Germany;2. CIRAD, UMR ASTRE, F-34398 Montpellier, France;3. ASTRE, Université de Montpellier, CIRAD, INRA, Montpellier, France;4. EID Mediterranée, 165 Avenue Paul Rimbaud, 34184 Montpellier, France;5. Pathogenesis and Control of Chronic Infections, Université de Montpellier, INSERM, EFS, Montpellier, France;6. Institut Pasteur, Biology of Infection Unit, Inserm U1117, Pathogen Discovery Laboratory, Paris, France;1. National Reference Centre for Arboviruses, Department of Microbiology, Medical School, Aristotle University of Thessaloniki, Thessaloniki, 54124 Greece;2. EcoDevelopment SA, Thessaloniki, Greece
Abstract:The epidemiology of West Nile virus (WNV) is unpredictable and changing. Availability of whole genome sequences enables the detailed molecular epidemiology studies and the evaluation and design of diagnostic tools. In the present study we provide two PCR-based protocols which can be applied directly on biological samples from hosts infected by WNV strains belonging to lineage 1 or lineage 2. It was shown that the protocols worked successfully even on samples with relatively low viral load.
Keywords:West nile virus  Next generation sequencing  Whole genome
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