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Journal of Clinical Microbiology, April 1999, p. 950-953, Vol. 37, No. 4
0095-1137/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Amplification of the Six Major Human Herpesviruses from Cerebrospinal Fluid by a Single PCR

Sophie Minjolle,1 C. Michelet,2 I. Jusselin,1 M. Joannes,3 F. Cartier,2 and R. Colimon1,*

Laboratoire de Bactériologie-Virologie1 and Service des Maladies Infectieuses,2 CHU Pontchaillou, Rennes, and Laboratoire Argène-Biosoft, Varilhes,3 France

Received 19 August 1998/Returned for modification 5 October 1998/Accepted 21 December 1998

We used a novel type of primer system, a system that uses stair primers, in which the primer sequences are based on consensus sequences in the DNA polymerase gene of herpesvirus to detect herpesviruses by PCR. A single PCR in a single tube detected the six major herpesviruses that infect the central nervous system: herpes simplex virus type 1 (HSV-1), and type 2 (HSV-2), cytomegalovirus (CMV), Epstein-Barr virus (EBV), varicella-zoster virus (VZV), and human herpesvirus 6 (HHV-6). We used the technique to analyze 142 cerebrospinal fluid (CSF) samples that had been stored at -80°C and compared the results with those obtained previously for the same samples by standard, targeted PCR. Four hundred one targeted PCR tests had been run with the 142 samples to detect HSV-1, HSV-2, CMV, and VZV; screening for EBV and HHV-6 was not prescribed when the samples were initially taken. Eighteen CSF samples tested positive by classic targeted PCR. The herpesvirus consensus PCR detected herpesviruses in 37 samples, including 3 samples with coinfections and 17 viral isolates which were not targeted. Two samples identified as infected by the targeted PCR tested negative by the consensus PCR, and eight samples that tested positive by the consensus PCR were negative by the targeted PCR. One hundred three samples scored negative by both the targeted and the consensus PCRs. This preliminary study demonstrates the value of testing for six different herpesviruses simultaneously by a sensitive and straightforward technique rather than screening only for those viruses that are causing infections as suggested by clinical signs.

* Corresponding author. Mailing address: Laboratoire de Bactériologie-Virologie, Faculté de Médecine, Université Rennes 1, 2 avenue du Pr Léon Bernard, CS 34317, 35 043 Rennes cedex, France. Phone: 0299284276. Fax: 0299284159. E-mail: colimon{at}mailhost.univ-rennes1.fr.

Journal of Clinical Microbiology, April 1999, p. 950-953, Vol. 37, No. 4
0095-1137/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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