©1996-2012 All Rights Reserved. Online Journal of Veterinary Research . You may not store these pages in any form except for your own personal use. All other usage or distribution is illegal under international copyright treaties. Permission to use any of these pages in any other way besides the before mentioned must be gained in writing from the publisher. This article is exclusively copyrighted in its entirety to OJVR.This article may be copied once but may not be, reproduced or re-transmitted without the express permission of the editors. This journal satisfies the refereeing requirements (DEST) for the Higher Education Research Data Collection (Australia). Linking:To link to this page or any pages linking to this page you must link directly to this page only here rather than put up your own page.
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)
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