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通过实时聚合酶链反应快速检测单纯疱疹病毒和水痘带状疱疹病毒感染

Rapid detection of herpes simplex virus and varicella-zoster virus infections by real-time PCR.

作者信息

Weidmann Manfred, Meyer-König Ursula, Hufert Frank T

机构信息

Department of Virology, Institute for Medical Microbiology and Hygiene, University of Freiburg, 79104 Freiburg, Germany.

出版信息

J Clin Microbiol. 2003 Apr;41(4):1565-8. doi: 10.1128/JCM.41.4.1565-1568.2003.

Abstract

The herpes simplex viruses types 1 and 2 (HSV-1 and HSV-2) and varicella-zoster virus (VZV) can cause life-threatening infections of the central nervous system and lead to severe infections in immunocompromised subjects and newborns. In these cases, rapid diagnosis is crucial. We developed three different real-time PCR assays based on TaqMan chemistry for the LightCycler instrument to detect HSV-1, HSV-2, and VZV. When the TaqMan assays were compared to our in-house nested PCR assays, the test systems had equal sensitivities of <or=10 plasmid copies per assay. When clinical samples were investigated by TaqMan PCR to detect HSV-1, HSV-2, and VZV DNA, 95, 100, and 96% of the samples determined to be positive by nested PCR, respectively, were positive by the real-time PCR assays. The specificities of all PCR assays were almost 100%. Furthermore, the TaqMan PCR assays could be performed within 2.5 h, whereas nested PCR results were available after 9 h. In addition to offering more rapid results, the TaqMan PCR assays appear to be less expensive than nested PCR assays due to less hands-on time. In summary, TaqMan PCR is an excellent alternative to conventional nested PCR assays for the rapid detection of HSV-1, HSV-2, and VZV in clinical samples.

摘要

1型和2型单纯疱疹病毒(HSV-1和HSV-2)以及水痘带状疱疹病毒(VZV)可引起危及生命的中枢神经系统感染,并导致免疫功能低下者和新生儿发生严重感染。在这些情况下,快速诊断至关重要。我们基于TaqMan化学原理为LightCycler仪器开发了三种不同的实时荧光定量PCR检测方法,用于检测HSV-1、HSV-2和VZV。当将TaqMan检测方法与我们的内部巢式PCR检测方法进行比较时,这些检测系统的灵敏度相同,每次检测均≤10个质粒拷贝。当通过TaqMan PCR检测临床样本以检测HSV-1、HSV-2和VZV DNA时,巢式PCR检测确定为阳性的样本中,分别有95%、100%和96%通过实时荧光定量PCR检测为阳性。所有PCR检测方法的特异性几乎均为100%。此外,TaqMan PCR检测可在2.5小时内完成,而巢式PCR结果在9小时后才能获得。除了能提供更快的结果外,由于操作时间较少,TaqMan PCR检测似乎比巢式PCR检测成本更低。总之,TaqMan PCR是临床样本中快速检测HSV-1、HSV-2和VZV的传统巢式PCR检测的极佳替代方法。

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Detection of Herpes simplex virus DNA by real-time PCR.通过实时聚合酶链反应检测单纯疱疹病毒DNA
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