Multiplex real-time RT-PCR detection of three viruses associated with the bovine respiratory disease complex
Multiplex real-time RT-PCR detection of three viruses associated with the bovine respiratory disease complex
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DOI：
10.1016/j.jviromet.2010.11.020
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发表时间：
2011-02-01
影响因子：
3.1
通讯作者：
Mahony, Timothy John
中科院分区：
文献类型：
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作者：
Horwood, Paul Francis;Mahony, Timothy John
牛呼吸道疾病（BRDC）被认为是北美乃至世界范围内与饲养场牛相关的最严重疾病。BRDC是一种多因素疾病，环境条件与多种病毒和细菌病原体相互作用，产生严重的呼吸系统疾病。牛疱疹病毒1型、牛病毒性腹泻病毒和牛副流感病毒3型是与BRDC相关的三种主要病毒。在这项研究中，利用Taqman引物和探针开发了一种多重实时RT-PCR，可以同时检测这三种重要的BRDC病毒。利用细胞培养感染材料和来自BRDC病例的牛临床样本对该检测方法进行了优化和验证。通过将多重检测与单个单一检测进行比较，分析了该检测的敏感性，结果表明，实时rt - pcr的多重检测不会抑制病毒检测。该技术是首次报道的BPIV-3的实时PCR方法，也是首次设计用于检测三种BRDC病毒病原体的多重PCR方法。皇冠版权所有(C) 2010由Elsevier B.V.出版。
Bovine respiratory disease complex (BRDC) is considered the most significant illness associated with feedlot cattle in North America and possibly worldwide. BRDC is a multi-factorial disease with environmental conditions interacting with multiple viral and bacterial pathogens to produce severe respiratory illness. Bovine herpesvirus 1, bovine viral diarrhoea virus and bovine parainfluenza virus 3 are three of the major viruses associated with BRDC. In this study, a multiplex real-time RT-PCR using Taqman primers and probes was developed to detect simultaneously all three of these important BRDC viruses. The assay was optimised and validated using cell-culture infected material and bovine clinical samples from BRDC cases. The sensitivity of the assay was analysed by comparing the multiplex with the individual singleplex assays, which demonstrated that viral detection was not inhibited, by multiplexing the real-time RT-PCRs. This technique is the first reported real-time PCR method for BPIV-3 and the first multiplex designed to detect three BRDC viral pathogens. Crown Copyright (C) 2010 Published by Elsevier B.V. All rights reserved.