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

摘要

Objectives: Herpes 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. Methods: DNA 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. Results: Compared 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. Conclusions: The 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.