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

摘要

Detection of African horsesickness virus (AHSV) by three different reverse transcription polymerase chainreaction (RT-PCR) based methods were compared. Primers were complementary to sequences contained in the geneencoding non-structural protein 2 of AHSV serotype 3. All three assays were group-specific for AHSV and detected allnine serotypes but not related orbiviruses. A dilution range was made of titrated chicken embryo-related cells infectedwith AHSV serotype 3 and, after isolating RNA from each diluted sample they were tested using the three differentassays. 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 Africanhorsesickness suspect field cases hemi-nested RT-PCR, intra-cerebral injection of mice and virus isolation from cellculture detected AHSV in 83%, 58% and 25% of samples respectively.