Development of real-time RT-PCR for the detection of low concentrations of Rift Valley fever virus

Marianne Maquart; Sarah Temmam; Jean-Michel Heraud; Isabelle Leparc-Goffart; Catherine Cetre-Sossah; Koussay Dellagi; Eric Cardinale; Hervé Pascalis

Development of real-time RT-PCR for the detection of low concentrations of Rift Valley fever virus

Marianne Maquart ^{1, 2, *} Sarah Temmam ^{1, 3} Jean-Michel Heraud ⁴ Isabelle Leparc-Goffart ⁵ Catherine Cetre-Sossah ^{2, 1} Koussay Dellagi ^{1, 6} Eric Cardinale ^{1, 2} Hervé Pascalis ^{1, 6}

* Corresponding author

1 CRVOI - Centre de Recherche et de Veille sur les Maladies Émergentes dans l'Océan Indien

2 UMR CMAEE - Contrôle des maladies animales exotiques et émergentes

3 LEM - Laboratoire d'Ecologie Microbienne - UMR 5557

4 IPM - Unité de Virologie [Antananarivo, Madagascar]

5 IRBA - Institut de Recherche Biomédicale des Armées [Antenne Marseille]

6 IRD - Institut de Recherche pour le Développement

Abstract : In recent years, Madagascar and the Comoros archipelago have been affected by epidemics of Rift Valley fever (RVF), however detection of Rift Valley fever virus (RVFV) in zebu, sheep and goats during the post epidemic periods was frequently unsuccessful. Thus, a highly sensitive real-time RT-PCR assay was developed for the detection of RVFV at low viral loads. A new RVF SYBR Green RT-PCR targeting the M segment was tested on serum from different RVF seronegative ruminant species collected from May 2010 to August 2011 in Madagascar and the Comoros archipelago and compared with a RVF specific quantitative real time RT-PCR technique, which is considered as the reference technique. The specificity was tested on a wide range of arboviruses or other viruses giving RVF similar clinical signs. A total of 38 out of 2756 serum samples tested positive with the new RT-PCR, whereas the reference technique only detected 5 out of the 2756. The described RT-PCR is an efficient diagnostic tool for the investigation of enzootic circulation of the RVF virus. It allows the detection of low viral RNA loads adapted for the investigations of reservoirs or specific epidemiological situations such as inter-epizootic periods.

Keywords : Diagnostic tool Inter-epidemic period Rift Valley fever virus Semi-nested RT-PCR

Document type :

Journal articles

Domain :

Life Sciences [q-bio] / Microbiology and Parasitology / Virology

Life Sciences [q-bio] / Santé publique et épidémiologie

Life Sciences [q-bio] / Bioengineering

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