Dried fluid spots for peste des petits ruminants virus load evaluation allowing for non-invasive diagnosis and genotyping
| Autor: | Mohammad Rafiqul Islam, Ataur Rahman Bhuiyan, Emmanuel Albina, Geneviève Libeau, Rokshana Parvin, Mushfiqur M Rahman, Olivier Kwiatek, Emdadul Haque Chowdhury |
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| Přispěvatelé: | Ministry of fisheries and livestock, Partenaires INRAE, Bangladesh Agricultural University, Contrôle des maladies animales exotiques et émergentes (UMR CMAEE), Centre de Coopération Internationale en Recherche Agronomique pour le Développement (Cirad)-Institut National de la Recherche Agronomique (INRA), Co-ordinated Research Project 'Development and validation of tests for rapid and specific diagnosis of Peste des Petits ruminants (PPR)', International Atomic Energy Agency (IAEA), EU Network of Excellence for Epizootic Disease Diagnosis and Control, (EPIZONE) |
| Jazyk: | angličtina |
| Předmět: |
Phylogénie
Veterinary medicine Genotyping Techniques [SDV]Life Sciences [q-bio] L73 - Maladies des animaux Genotype media_common Filtre Spots Goats Peste des petits ruminants virus General Medicine Viral Load 3. Good health ARN Nasal Swab Field samples Viral load Génotype Research Article Genotyping Testage non destructif Biology Real-Time Polymerase Chain Reaction Filter paper Virus peste petits ruminants Peste-des-petits-ruminants virus Peste-des-Petits-Ruminants Animals media_common.cataloged_instance Surveillance épidémiologique Diagnostic European union Technique analytique Goat Diseases General Veterinary veterinary(all) Dried Blood Spot Testing U30 - Méthodes de recherche |
| Zdroj: | BMC Veterinary Research BMC Veterinary Research, BioMed Central, 2014, 10, ⟨10.1186/s12917-014-0247-y⟩ BMC Veterinary Research (10), . (2014) |
| ISSN: | 1746-6148 |
| DOI: | 10.1186/s12917-014-0247-y |
| Popis: | International audience; Background: Active surveillance of peste des petits ruminants (PPR) should ease prevention and control of this disease widely present across Africa, Middle East, central and southern Asia. PPR is now present in Turkey at the gateway to the European Union. In Bangladesh, the diagnosis and genotyping of PPR virus (PPRV) may be hampered by inadequate infrastructures and by lack of proper clinical material, which is often not preserved under cold chain up to laboratories. It has been shown previously that Whatman (R) 3MM filter paper (GE Healthcare, France) preserves the nucleic acid of PPRV for at least 3 months at 32 degrees C. Results: In this study, we demonstrate the performances of filter papers for archiving RNA from local PPRV field isolates for further molecular detection and genotyping of PPRV, at -70 degrees C combined with ambient temperature, for periods up to 16 months. PPR-suspected live animals were sampled and their blood and nasal swabs were applied on filter papers then air dried. Immediately after field sampling, RT-PCR amplifying a 448-bp fragment of the F gene appeared positive for both blood and nasal swabs when animals were in febrile stage and only nasal swabs were detected positive in non-febrile stage. Those tested positive were monitored by RT-PCR up to 10 months by storage at -70 degrees C. At 16 months, using real time RT-PCR adapted to amplify the N gene from filter paper, high viral loads could still be detected (similar to 2 x 10(7) copy numbers), essentially from nasal samples. The material was successfully sequenced and a Bayesian phylogenetic reconstruction achieved adequate resolution to establish temporal relationships within or between the geographical clusters of the PPRV strains. Conclusions: This clearly reveals the excellent capacity of filter papers to store genetic material that can be sampled using a non-invasive approach. |
| Databáze: | OpenAIRE |
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