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Identification and differentiation of the nine African horse sickness virus serotypes by RT–PCR amplification of the serotype-specific genome segment 2

Agence Française de Sécurité Sanitaire des Aliments – Alfort, Laboratoire Central de Recherches Vétérinaires, 22 rue Pierre Curie, 94703 Maisons-Alfort, France¹
Institute for Animal Health, Pirbright Laboratory, Ash Road, Pirbright, Woking, Surrey GU24 0NF, UK²
Onderstepoort Veterinary Institute, Private Bag X5, Onderstepoort 0110, Republic of South Africa³

Author for correspondence: Corinne Sailleau. Fax +33 1 43 68 97 62. e-mail c.sailleau{at}alfort.afssa.fr

This paper describes the first RT–PCR for discrimination of the nine African horse sickness virus (AHSV) serotypes. Nine pairs of primers were designed, each being specific for one AHSV serotype. The RT–PCR was sensitive and specific, providing serotyping within 24 h. Perfect agreement was recorded between the RT–PCR and virus neutralization for a coded panel of 56 AHSV reference strains and field isolates. Serotyping was achieved successfully with live and formalin-inactivated AHSVs, with isolates of virus after low and high passage through either tissue culture or suckling mouse brain, with viruses isolated from widely separated geographical areas and with viruses isolated up to 37 years apart. Overall, this RT–PCR provides a rapid and reliable method for the identification and differentiation of the nine AHSV serotypes, which is vital at the start of an outbreak to enable the early selection of a vaccine to control the spread of disease.

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