Evaluation of Reverse Transcription-pcr Protocols Based on the Fusion Gene for Diagnosis of Bovine Respiratory Syncytial Virus Infections
نویسندگان
چکیده
Bovine respiratory syncytial virus (BRSV) is a pneumovirus in the family paramyxoviridae, is an important cause of acute respiratory disease in postweaning calves and feedlot cattle. The real-time reverse transcriptase PCR protocols were developed to detect BRSV infection in infected animals. The sensitivity of RT-PCR protocols based on fusion gene were evaluated using different Mastermixes such as Qiagen One Step RT-PCR (Qiagen) for conventional RT-PCR, Superscrip probe (Invitrogen) and QuentiTec probe (Qiagen) for realtime RT-PCR with and without internal control. The detection limit of different RT-PCR protocols using serial dilutions from BRSV plasmid and based on different probes was 10 RNA copies/ml. Furthermore, the specificity of real-time RT-PCR was evaluated using different bacterial and viral strains which can be isolated from respiratory infected animals. In another side, the real-time RT-PCR in combination with ß-actin and conventional RT-PCR showed detectable Ctvalues only with BRSV strain.
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