A Comparison of Different PCR-Based Methods for the Detection of African Horsesickness Virus

ABSTRACT

Detection of African horsesickness virus (AHSV) by three different reverse transcription polymerase chain reaction (RT-PCR) based methods were compared. Primers were complementary to sequences contained in the gene encoding non-structural protein 2 of AHSV serotype 3. All three assays were group-specific for AHSV and detected all nine serotypes but not related orbiviruses. A dilution range was made of titrated chicken embryo-related cells infected with AHSV serotype 3 and, after isolating RNA from each diluted sample they were tested using the three different assays. RNA representing 0.2, 2 and 20 plaque forming units of AHSV could be detected by hemi-nested RT-PCR, PCRenzyme- linked immunosorbent assay (PCR-ELISA) and RT-PCR respectively. In twelve samples from African horsesickness suspect field cases hemi-nested RT-PCR, intra-cerebral injection of mice and virus isolation from cell culture detected AHSV in 83%, 58% and 25% of samples respectively.

Keywords:

African horsesickness virus (AHSV), polymerase chain reaction (PCR), reverse transcription PCR (RT-PCR), hemi-nested PCR, PCR-ELISA.