Rahaus Markus, Desloges Nathalie, Wolff Manfred H
Institute of Microbiology and Virology, University of Witten/Herdecke, Stockumer Str. 10, D-58448, Witten, Germany.
J Virol Methods. 2003 Feb;107(2):257-60. doi: 10.1016/s0166-0934(02)00250-1.
The reverse transcription polymerase chain reaction (RT-PCR) is used commonly to analyse transcription of genes. In the field of virology it is an extremely helpful method to analyse the transcriptional activity of both, DNA and RNA viruses. The standard RT-PCR allows an investigation of the activity of only one gene. Here, we describe the development of a sensitive multiplex RT-PCR assay for single tube amplification to analyse a set of different genes encoded by the varicella-zoster virus (VZV), a member of the human-pathogenous herpesviruses. This multiplex RT-PCR amplifies the genes ORF 4, ORF 21 and ORF 68; each of these three genes belongs to a distinct class of genes, which is expressed one after the other within the infectious cycle. This method provides the possibility for rapid but extensive examination of VZV transcriptional activity and could be used in both fields, fundamental research as well as clinical diagnostic work.
逆转录聚合酶链反应（RT-PCR）通常用于分析基因转录。在病毒学领域，它是分析DNA和RNA病毒转录活性的一种极其有用的方法。标准RT-PCR仅能检测一个基因的活性。在此，我们描述了一种用于单管扩增的灵敏多重RT-PCR检测方法的开发，以分析由人类致病性疱疹病毒之一的水痘带状疱疹病毒（VZV）编码的一组不同基因。这种多重RT-PCR扩增ORF 4、ORF 21和ORF 68基因；这三个基因中的每一个都属于不同的基因类别，它们在感染周期内依次表达。该方法为快速但广泛地检测VZV转录活性提供了可能，可用于基础研究和临床诊断工作这两个领域。
