Using a new serotype-specific Polymerase Chain Reaction (PCR) and sequencing to differentiate between field and vaccine-derived African Horse Sickness viruses submitted in 2016/2017

Abstract

The outer capsid viral protein 2 (VP2) of African horse sickness virus, encoded by the most variable genome segment 2 (Seg-2), is the primary target for AHSV-specific neutralising antibodies and thus determines the virus serotype. Full length segment 2 sequences from more than 100 AHSVs isolated over the last 80 years were compared and single nucleotide polymorphisms (SNPs) identified between the reference strains and recent field viruses. Regions unique to each individual serotype were identified and primers designed to differentially amplify each of the nine serotypes. The sequences of resulting amplicons contained a significant amount of SNPs to discriminate between field viruses and reference strains or live attenuated viruses. The new serotype specific RT-PCR were subsequently used to determine the prevalence of different AHSV serotypes associated with samples submitted to the Agricultural Research Council - Onderstepoort Veterinary Research Institute during the 2016 / 2017 season. Subsequent sequencing of the PCR products were used to determine if the infections were caused by field or vaccine-derived strains. The serotypes of 70 AHSV positive diagnostic samples submitted to the ARC-OVR were determined. Serotypes 2 and 6 were the most prevalent, while Serotype 1 was the only serotype where sequences identical to the ALV or reference strains were detected in field samples. Based on this study, the incidence of vaccine-derived AHS infections submitted from southern Africa were low. This serotype-specific RT-PCR and sequencing assay could assist with the surveillance and control of equines movement nationally and internationally. It could also provide valuable scientific guidance on the policies and guidelines regulating vaccination and trade of equines in South Africa.