Markoulatos P, Georgopoulou A, Kotsovassilis C, Karabogia-Karaphillides P, Spyrou N
Hellenic Pasteur Institute, Department of Virology, Athens, Greece.
J Clin Lab Anal. 2000;14(5):214-9. doi: 10.1002/1098-2825(2000)14:5<214::AID-JCLA3>3.0.CO;2-4.
The development of a multiplex polymerase chain reaction method for the rapid and accurate detection and typing of HSV-1, HSV-2, and VZV from clinical specimens is described. A sensitive multiplex polymerase chain reaction was achieved by optimization of parameters such as the primers, magnesium, and dNTPs concentrations. False-negative results that sometimes arise due to inhibitors of DNA amplification or failure of DNA extraction procedure used may be avoided by assaying each specimen with alpha-tubulin primers. Multiplex PCR amplified viral sequences from all 55 specimens obtained from patients with clinical evidence of HSV or VZV infection indicated 100% sensitivity. From 55 patients who were investigated by multiplex PCR, HSV-1 was detected in 28, HSV-2 in 20, and VZV in 7 specimens. The reported results indicate that the present multiplex PCR assay has a potential application in clinical diagnosis when a rapid and accurate detection and typing of involved viruses HSV-1, HSV-2, or VZV is needed.
本文描述了一种用于从临床标本中快速、准确地检测和分型单纯疱疹病毒1型（HSV-1）、单纯疱疹病毒2型（HSV-2）和水痘带状疱疹病毒（VZV）的多重聚合酶链反应方法的开发。通过优化引物、镁离子和脱氧核苷三磷酸（dNTPs）浓度等参数，实现了灵敏的多重聚合酶链反应。通过使用α-微管蛋白引物检测每个标本，可以避免因DNA扩增抑制剂或所用DNA提取程序失败而有时出现的假阴性结果。对所有55份来自有HSV或VZV感染临床证据患者的标本进行多重聚合酶链反应扩增病毒序列，结果显示灵敏度为100%。在接受多重聚合酶链反应检测的55名患者中，28份标本检测到HSV-1，20份检测到HSV-2，7份检测到VZV。报告结果表明，当需要对涉及的病毒HSV-1、HSV-2或VZV进行快速、准确的检测和分型时，目前的多重聚合酶链反应检测法在临床诊断中具有潜在应用价值。
