Detection of the four major human herpesviruses simultaneously in whole blood and cerebrospinal fluid samples by the fluorescence polarization assay

Abstract

SummaryObjectivesHerpes simplex virus type 1/2 (HSV-1/-2), cytomegalovirus (CMV), and Epstein–Barr virus (EBV) correlate strongly with infections of the central nervous system. The objective of this study was to develop a method for the simultaneous detection of HSV-1/-2, CMV, and EBV DNA by the fluorescence polarization assay based on asymmetric polymerase chain reaction (PCR) and hybridization.MethodsDNA of HSV-1/-2, CMV, and EBV was amplified in an asymmetric PCR by a universal primer system. The amplicons were then detected by the fluorescence polarization assay. In this method, the probes for HSV-1/-2, CMV, and EBV hybridized with their respective target amplicons, and the hybridization resulted in an increase in the fluorescence polarization values. Infections of HSV-1/-2, CMV, and EBV were determined by the increased fluorescence polarization values. The DNA extracted from whole blood and cerebrospinal fluid samples was subjected to fluorescence polarization and a previously published multiplex PCR assay in parallel.ResultsCompared to the multiplex PCR assay, no significant difference in the numbers of samples positive for the human herpesviruses was identified by the fluorescence polarization assay.ConclusionsThe fluorescence polarization assay presented in this study is a reliable, convenient, and cost-effective diagnostic tool that allows the detection of the four major human herpesviruses