Analysis of Human Cytomegalovirus Using the Polymerase Chain Reaction

As with numerous other branches of science, the study of human cytomegalovirus (HCMV) infection has been revolutionized by the polymerase chain reaction (
PCR
) method first devised by Mullis and Faloona (
1
). PCR allows the in vitro amplification of HCMV
DNA
sequences by the simultaneous primer extension of complementary DNA strands. Similarly, reverse transcription-PCR (
RT-PCR
) allows the study of targeted gene expression, by reverse transcription of RNA to complementary DNA (
cDNA
), followed by amplification of target DNA using predetermined primers. The PCR method is used in the clinical diagnosis of HCMV infection, particularly in the setting of transplantation medicine and in those patients infected with the human immunodeficiency virus (HIV). In addition, the advent of PCR and RT-PCR has transformed our understanding of the pathogenesis of HCMV infection, central to which is the definition of the sites of latency, the degree and type of gene expression within the latently infected cell, and the factors influencing both the maintenance of latency and reactivation of the virus during immunosuppression.