Appling real time RT-PCR for bluetongue virus detection in Iran



During 2009-10, real time RT-PCR and conventional RT-PCR techniques Used for detecting BTVs RNA in
310 blood samples. For real time and gel based RT-PCR segment-1 and segment-10 selected as conserve
genes to search any BTV strains. Using these methods, 58 (%18.7) and 14 (%4.5) positive samples were
detected among the clinically suspected sheep. Sensitivity of both molecular techniques evaluated by log-10
serial dilutions of BTV16 RNA, and determined 101.8 and 103.8 TCID50/ml in rRT-PCR and conventional
RT-PCR respectively. This report confirmed rRT-PCR assay could detect weak BTV positive samples even
at end stage of infection. In this study Virus isolation from selected positive samples failed by inoculation to
embryonated chicken egg, Vero and KC cell.