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Journal of Clinical Microbiology, September 2000, p. 3274-3279, Vol. 38, No. 9
0095-1137/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Comprehensive PCR-Based Assay for Detection and Species Identification of Human Herpesviruses

Grant Johnson, Susan Nelson,dagger Martin Petric, and Raymond Tellier*

Department of Paediatric Laboratory Medicine, The Hospital for Sick Children, Toronto, Ontario, Canada

Received 14 March 2000/Returned for modification 12 May 2000/Accepted 26 June 2000

The description and evaluation of a PCR-based assay for the detection and species identification of the eight known human herpesviruses are presented. Two primer pairs targeting well-conserved regions of the genome allowed the amplification of the DNAs of all known human herpesviruses at a high level of sensitivity (10 to 100 genome copies for most viruses). Identification of the virus species was achieved through restriction enzyme digestion with BamHI and BstUI, which yielded fragment sizes that were characteristic for each herpesvirus. Furthermore, it was demonstrated that this restriction enzyme panel allowed the discrimination between human herpesvirus 6 variant A and variant B. This assay format was validated over the course of 1 year in a clinical virology laboratory setting, where it was shown that it readily detected human herpesviruses, including occasional multiple infections, in a variety of clinical samples. The PCR assay was compared to isolation and electron microscopy for the detection of herpes simplex (HSV) and varicella-zoster virus (VZV) in clinical samples. All specimens positive by conventional methods were also positive by PCR. However, in a number of clinical specimens in which HSV or VZV could not be detected by conventional methods, PCR was able to demonstrate the presence of the virus.

* Corresponding author. Mailing address: Department of Paediatric Laboratory Medicine, The Hospital for Sick Children, 555 University Ave., Toronto, Ontario, Canada M5G 1X8. Phone: (416) 813-6592. Fax: (416) 813-6257. E-mail: raymond.tellier{at}sickkids.on.ca.

dagger Present address: Aventis Pasteur Canada.

Journal of Clinical Microbiology, September 2000, p. 3274-3279, Vol. 38, No. 9
0095-1137/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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