Evaluation of RT-PCR for detection of Sudanese serotypes of epizootic hemorrhagic disease virus serogroup

Epizootic hemorrhagic disease (EHD) is an infectious non-contagious disease of deer and cattle. At least 2 serotypes of EHD virus, designated EHDV-4 and EHDV-318, are enzootic in the Sudan. To facilitate clinical disease investigation and control of the disease a rapid diagnostic assay is urgently needed. A reverse transcriptase (RT) polymerase chain reaction (RT-PCR) protocol, previously reported for detection of the United States EHDV serotypes 1 and 2 ribonucleic acid (RNA) in cell culture and clinical specimens, was evaluated for detection of the Sudanese EHDV serotypes. RNA from Sudanese isolates of EHDV-4 and EHDV-318, propagated in cell cultures, were detected by the described RT-PCR-based assay. The specific 387 bp PCR products were visualized on ethidium bromide-stained agarose gel. Amplification product was not detected when the EHDV RT-PCR-based assay was applied to RNA from Sudanese bluetongue virus (BTV) serotype 4 (BTV-4) or total nucleic acid extracts from uninfected BHK-21 cells. The scientific observations reported in this paper indicated that the previously described EHDV RT-PCR assay could be applied for detection of EHDV infection among the Sudanese susceptible animal populations.