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

Abstract

During 2009-10, real time RT-PCR and conventional RT-PCR techniques Used for detecting BTVs RNA in310 blood samples. For real time and gel based RT-PCR segment-1 and segment-10 selected as conservegenes to search any BTV strains. Using these methods, 58 (%18.7) and 14 (%4.5) positive samples weredetected 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.

Keywords

• Real time RT-PCR
• Conventional RT-PCR
• Bluetongue