Detection of the four major human herpesviruses simultaneously in whole blood and cerebrospinal fluid samples by the fluorescence polarization assay.
OBJECTIVE
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.
Zhongliang
Wu
W
Department of Neurology, Xijing Hospital, The Fourth Military Medical University, Xian, China.
Rui
Chen
C
Xiangling
Wang
W
Ding
Li
L
Jinrong
Zhao
Z
Yanhai
Guo
G
Ju
Zhang
Z
D004279:DNA, Viral / Q000097:blood / Q000134:cerebrospinal fluid; D005454:Fluorescence Polarization / Q000379:methods*; D006564:Herpesviridae / Q000302:isolation & purification*; D006566:Herpesviridae Infections / Q000097:blood / Q000134:cerebrospinal fluid / Q000175:diagnosis*; D006801:Humans; D009693:Nucleic Acid Hybridization; D016133:Polymerase Chain Reaction; D012680:Sensitivity and Specificity
D004279:DNA, Viral