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一种用于快速鉴定和半定量麻疹病毒的实时荧光定量PCR检测方法的开发与评估

Development and evaluation of a real-time PCR assay for rapid identification and semi-quantitation of measles virus.

作者信息

Thomas Brenda, Beard Stuart, Jin Li, Brown Kevin E, Brown David W G

机构信息

Virus Reference Department, Centre for Infections, Health Protection Agency, London, UK.

出版信息

J Med Virol. 2007 Oct;79(10):1587-92. doi: 10.1002/jmv.20997.

Abstract

A real-time PCR assay for measles virus was designed and validated using clinical samples including oral fluids, sera, urines, throat swabs, blood samples, and nasopharyngeal aspirates. The test was specific for measles virus, with a slightly higher sensitivity compared to the conventional nested PCR. Calculation of viral genome number in these samples, by comparison with a standard curve prepared from dilutions of cloned measles virus H gene, indicated that, overall, serum samples tended to have a lower viral load than oral fluid samples, and that the viral load decreased with increasing time after onset of symptoms. The real-time PCR is considered to be a sensitive and specific alternative to the conventional measles PCR, especially in situations where early and rapid diagnosis are important.

摘要

设计了一种用于检测麻疹病毒的实时聚合酶链反应(PCR)检测方法,并使用包括口腔液、血清、尿液、咽拭子、血液样本和鼻咽抽吸物在内的临床样本进行了验证。该检测方法对麻疹病毒具有特异性,与传统巢式PCR相比,灵敏度略高。通过与由克隆的麻疹病毒H基因稀释液制备的标准曲线进行比较,计算这些样本中的病毒基因组数量,结果表明,总体而言,血清样本的病毒载量往往低于口腔液样本,并且病毒载量随着症状出现后时间的增加而降低。实时PCR被认为是传统麻疹PCR的一种敏感且特异的替代方法,尤其是在早期快速诊断很重要的情况下。

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