Rt-Pcr Detection of African Horse Sickness Virus Serogroups In Cell Culture

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Elgenay , Samah Abdelrahman Ahmed
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University of Khartoum
In the present work, a reverse transcriptase polymerase chain reaction (RT-PCR) protocol was carried out to detect African horse sickness virus (AHSV) RNA in vero cell culture. The nine serotypes of African horse sickness virus were used in this study . All these serotypes werepropagated in vero cell culture. RNAs were extracted from these serotypes and then, they were detected by the described RT-PCR assay, using primers derived from segment 3 of AHSV serotype 6. The specific 240 bp PCR products were amplified from all AHSV serotypes used in the study,and they were visualized on ethidium bromide-stained agarose gel. The Amplification product was not detected when the RT-PCR assay was applied to RNA from bluetongue virus (BTV) or palyam virus. This specific 240 bp PCR product was obtained from an mount of 10 pg ,1.0 pg, 100fg, RNA from AHSV serotype (6). The results of this study showed that the described RT-PCR assay, using the mentioned primers, could be applied for detection of AHSV serotypes. X In conclusion, the AHSV RT- PCR can solve the problems which are found in virus isolation procedures. The rapidity, sensitivity and specificity of the RT- PCR assay would greatly facilitate detection of AHSV infection in an out break among susceptible equine.
71 Pages
Chicken; Enzyme; protein; Blood; water; Sodium ; Africa