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OJVRTM
Online
Journal of Veterinary Research©
Volume 16 (2):62-71, 2012. Slightly Redacted 2017.
Reverse transcription-PCR to detect Bovine
Respiratory Syncytial Virus (BRSV)
1Faculty of Veterinary Medicine, Benha University, Mostohor, Toukh, Egypt (*Leipzig, Germany)
SUMMARY
Selim A., Reverse transcription-PCR to detect bovine respiratory
syncytial virus (BRSV), Onl J Vet Res., 16 (2):62-71, 2012. BRSV is an important cause of acute
respiratory disease mostly in post weaning calves and feedlot cattle but also
in adult cattle. Real-time reverse
transcriptase PCR protocols were developed to detect BRSV
infection in infected animals. Sensitivity of RT-PCR protocols targeting fusion
gene was optimized using different Mastermixes such
as Qiagen One Step RT-PCR Kit (Qiagen)
for conventional RT-PCR, Superscript probe (Invitrogen) and QuantiTect
probe (Qiagen) for real-time 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. The specificity of real-time RT-PCR was
evaluated using different bacterial and viral strains which could be isolated
from respiratory infected animals. Real-time RT-PCR in combination with
ß-actin and conventional RT-PCR showed detectable Ct-values only with
BRSV strain.
Keywords: BRSV, real-time RT-PCR, conventional
PCR, internal control
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