DEVELOPMENT AND EVALUATION OF A MULTIPLEX CLASSICAL RT-PCR FOR SIMULTANEOUS DETECTION AND TYPING OF FMDV IN WEST AFRICA - Anses - Agence nationale de sécurité sanitaire de l’alimentation, de l’environnement et du travail Accéder directement au contenu
Communication Dans Un Congrès Année : 2018

DEVELOPMENT AND EVALUATION OF A MULTIPLEX CLASSICAL RT-PCR FOR SIMULTANEOUS DETECTION AND TYPING OF FMDV IN WEST AFRICA

Kamila Gorna
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Eve Laloy
Anthony Relmy
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Aurore Romey
Stéphan Zientara
Sandra Blaise-Boisseau
Labib Bakkali Kassimi

Résumé

Introduction: Foot-and-Mouth Disease, (FMD) caused by Foot-and–Mouth Disease Virus (FMDV) is one of the most economically devastating diseases affecting cloven-hoofed animals. There are seven immunologically and genetically distinct serotypes (O/A/C/Asia1/SAT1-3) further subdivided into different topotypes and genotypes. The West African territories are considered as regions with continuous FMDV circulation of serotypes O, A, SAT1 and SAT2. An early diagnosis of FMD is crucial to implement adequate outbreak management. This study describes the development of a multiplex conventional RT-PCR for both detection and typing of FMD virus circulating in this region. Materials-methods: The RT-PCR reactions were developed with primer sets targeting the 3D coding region, the VP1 coding region (O/A/SAT1/SAT2-specific) and the -actin gene to produce amplicons with sizes easily distinguishable by agarose electrophoresis. A 6-plex prototype (O/A/SAT1/SAT2/3D/-actin) was finally developed using a panel of reference strains and positive field samples from Benin and the corresponding isolates. This test is currently evaluated on a larger panel of field samples collected in Nigeria and Senegal. Results: Using 6-plex protocol, 37 samples from Benin were positive for the 3D target and were correctly serotyped by 6-plex (33) or by 3-plex and simplex RT-PCR (4). 39/40 isolates from Nigeria were properly serotyped using 6-plex (30) or simplex (9). The corresponding field samples as well 39 clinical positive samples from Senegal are under investigation. Discussion-Conclusions: We have developed and evaluated a 6-plex RT-PCR that could be easily implemented in diagnostic laboratories in endemic countries, providing an improvement for rapid detection and typing of FMDV strains.
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anses-04441892 , version 1 (06-02-2024)

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  • HAL Id : anses-04441892 , version 1

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Kamila Gorna, Gaye Laye Diop, Marianne Diop, Eve Laloy, Modou Moustapha Lo, et al.. DEVELOPMENT AND EVALUATION OF A MULTIPLEX CLASSICAL RT-PCR FOR SIMULTANEOUS DETECTION AND TYPING OF FMDV IN WEST AFRICA. 5TH CONGRESS OF THE EUROPEAN ASSOCIATION OF VETERINARY DIAGNOSTICIANS (EAVLD 2018), EUROPEAN ASSOCIATION OF VETERINARY DIAGNOSTICIANS, Oct 2018, Bruxelles (BEL), Belgium. ⟨anses-04441892⟩
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