Development of a multiplex RT-PCR to detect transcription of varicella-zoster virus encoded genes.

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.