Abstract
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 ?-actin and conventional RT-PCR showed detectable Ctvalues only with BRSV strain.
Highlights
Bovine respiratory syncytial virus (BRSV) is a fragile RNA, a singlestranded negative-sense RNA virus which belongs to the pneumovirus genus, a member of the Paramyxoviridae family
BRSV appears to be an important virus in the bovine respiratory disease complex because of its frequency of occurrence, predilection for the lower respiratory tract, and its ability to predispose the respiratory tract to secondary bacterial infection (Ames, 1993)
PCR efficiency of real-time RT-PCR was evaluated with QuantiTect probe and with Superscript probe based on serial dilution of virus control strain PK 1/06 and for duplex real-time RT-PCR with QuentiTec probe
Summary
Bovine respiratory syncytial virus (BRSV) is a fragile RNA, a singlestranded negative-sense RNA virus which belongs to the pneumovirus genus, a member of the Paramyxoviridae family. Its genome has approximately 15140 nucleotides and encodes 10 different mRNA molecules (Buchholz et al, 1999; Grubbs et al, 2001). These viruses may cause respiratory tract infection and disease but often predispose the cattle to the bacterial pathogens. BRSV appears to be an important virus in the bovine respiratory disease complex because of its frequency of occurrence, predilection for the lower respiratory tract, and its ability to predispose the respiratory tract to secondary bacterial infection (Ames, 1993). The virus is transmitted when an animal that is infected sheds the virus in secretions such as nasal discharge and a common example of this would be nose to nose contact (Knight et al, 2001). BRSV infection causes severe respiratory signs in young cattle and frequently leads to the death of the infected animal (Baker et al, 1997)
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