Weidmann Manfred, Meyer-König Ursula, Hufert Frank T
Department of Virology, Institute for Medical Microbiology and Hygiene, University of Freiburg, 79104 Freiburg, Germany.
J Clin Microbiol. 2003 Apr;41(4):1565-8. doi: 10.1128/JCM.41.4.1565-1568.2003.
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.
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。