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OJVRTM
Online Journal of Veterinary Research©
Volume
16 (2):62-71, 2012
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),
Online 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