A reverse transcriptase (RT) polymerase chain reaction (RT-PCR)-based assay, for detection of African isolates of palyam virus ribonucleic acid (RNA) in cell culture, was developed. A pair of oligoribonucleotide primers (pal1 and pal2), selected from genome segment 3 of Chuzan virus, an isolate of the palyam serogroup, was used as a target for PCR amplification. Using RT-PCR, the pair of primers (pal1 and pal2) resulted in amplification of a 660-bp product. RNA samples from African isolates of palyam virus serogroup, propagated in cell cultures, were detected by this RT-PCR-based assay. Amplification product was not detected when the palyam RT-PCR-based assay was applied to RNA from, closely related orbiviruses, bluetongue virus (BTV) and epizootic hemorrhagic disease virus (EHDV); total nucleic acid extracts from uninfected Vero cells. The described RT-PCR-based assay provides a rapid, sensitive and specific method for detection and differentiation of palyam serogroup of orbiviruses in cell culture.
Abdel Rahim E. Karrar , Mohamed A. Abdalla , Ali M. Majid and Mohammed M. M. Salih . Application of RT-PCR for Detection of African Isolates of Palyam Orbiviruses Serogroup in Cell Culture.
DOI: https://doi.org/10.36478/javaa.2003.671.674
URL: https://www.makhillpublications.co/view-article/1680-5593/javaa.2003.671.674