Performance of real-time reverse transcription polymerase chain reaction for the detection of Foot-and-mouth disease virus during field outbreaks in the United Kingdom in 2007
Performance of real-time reverse transcription polymerase chain reaction for the detection of Foot-and-mouth disease virus during field outbreaks in the United Kingdom in 2007
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DOI：
10.1177/104063870902100303
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发表时间：
2009-05-01
影响因子：
1.5
通讯作者：
King, Donald P.
中科院分区：
文献类型：
--
作者：
Reid, Scott M.;Ebert, Katja;King, Donald P.
快速准确的诊断是有效控制口蹄疫的关键。本报告介绍了为采用实时逆转录聚合酶链式反应(实时逆转录聚合酶链式反应)处理2007年英国口蹄疫暴发期间收到的样本而采取的实际步骤。针对口蹄疫病毒(FMDV)基因组不同区域(5‘非编码区和3D区)的两种独立的实时RT-PCR方法被用来证实从第一个感染场所采集的临床样本中存在口蹄疫病毒。一旦对负责的口蹄疫病毒毒株进行了测序，就选择了一种单一的实时RT-PCR分析(3D)来测试总共3216个样本，包括来自所有8个受感染场所的材料。使用96孔自动化系统制备核酸模板，在提交后5小时内可处理84个样本，每个工作日测试多达269个样本。使用保守的截止值来指定阳性样本，对于阴性对照材料或从阴性场所收集的样本，其检测特异性分别为99.9%或100%。第一次，实时RT-PCR结果被用于识别牛群的临床前口蹄疫。此外，在疫情暴发的后期阶段，实时RT-PCR检测支持在高风险牛群中开展积极的监测计划。据作者所知，这是在将口蹄疫引入一个没有口蹄疫(未接种疫苗)的国家后，首次将实时RT-PCR作为主要实验室诊断工具的文件使用，并突出了这种分析在疾病暴发期间支持控制决策的优势。
Rapid and accurate diagnosis is essential for effective control of foot-and-mouth disease (FM D). The present report describes the practical steps undertaken to deploy a real-time reverse transcription polymerase chain reaction (real-time RT-PCR) to process the samples received during the Outbreaks of FMD in the United Kingdom in 2007. Two independent real-time RT-PCR assays targeting different regions (5' UTR and 3D) of the FMD virus (FMDV) genome were used to confirm the presence of FMDV in clinical samples collected from the first infected premises. Once the FMDV strain responsible had been sequenced, a single real-time RT-PCR assay (3D) was selected to test a total of 3,216 samples, including material from all 8 infected premises. Using a 96-well automated system to prepare nucleic acid template, tip to 84 samples Could be processed within 5 hr of Submission, and up to 269 samples were tested per working day. A conservative cut-off was used to designate positive samples, giving rise to an assay specificity of 99.9% or 100% for negative control material or samples collected from negative premises, respectively. For the first time, real-time RTPCR results were used to recognize preclinical FMD in a cattle herd. Furthermore, during the later stages of the outbreaks, the real-time RT-PCR assay Supported an active Surveillance program within high-risk cattle herds. To the authors' knowledge, this is the First documented use of real-time RT-PCR as a principal laboratory diagnostic tool following introduction of FMD into a Country that was FMD-free (without vaccination) and highlights the advantages of this assay to support control decisions during disease outbreaks.