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Journal of Clinical Microbiology, April 2004, p. 1409-1413, Vol. 42, No. 4
0095-1137/04/$08.00+0     DOI: 10.1128/JCM.42.4.1409-1413.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Detection and Genotyping of Varicella-Zoster Virus by TaqMan Allelic Discrimination Real-Time PCR

Paul A. Campsall,1,2 Nicholas H. C. Au,1 Julie S. Prendiville,3 David P. Speert,2 Rusung Tan,1 and Eva E. Thomas1*

Department of Pathology and Laboratory Medicine,1 Division of Pediatric Dermatology, Department of Pediatrics, University of British Columbia and Children's & Women's Health Centre of British Columbia,3 Division of Infectious and Immunological Diseases, Department of Pediatrics and the British Columbia Research Institute for Children's and Women's Health, University of British Columbia, Vancouver, British Columbia, Canada2

Received 15 September 2003/ Returned for modification 21 October 2003/ Accepted 6 November 2003

A proportion of individuals vaccinated with live attenuated Oka varicella-zoster virus (VZV) vaccine subsequently develop attenuated chicken pox and/or herpes zoster. To determine whether postvaccination varicella infections are caused by vaccine or wild-type virus, a simple method for distinguishing the vaccine strain from wild-type virus is required. We have developed a TaqMan real-time PCR assay to detect and differentiate wild-type virus from Oka vaccine strains of VZV. The assay utilized two fluorogenic, minor groove binding probes targeted to a single nucleotide polymorphism in open reading frame 62 that distinguishes the Oka vaccine from wild-type strains. VZV DNA could be genotyped and quantified within minutes of thermocycling completion due to real-time monitoring of PCR product formation and allelic discrimination analysis. The allelic discrimination assay was performed in parallel with two standard PCR-restriction fragment length polymorphism (RFLP) methods on 136 clinical and laboratory VZV strains from Canada, Australia, and Japan. The TaqMan assay exhibited a genotyping accuracy of 100% and, when compared to both PCR-RFLP methods, was 100 times more sensitive. In addition, the method was technically simpler and more rapid. The TaqMan assay also allows for high-throughput genotyping, making it ideal for epidemiologic study of the live attenuated varicella vaccine.

* Corresponding author. Mailing address: Children's and Women's Health Center of British Columbia, 4500 Oak St., Vancouver, BC V6H 3V4, Canada. Phone: (604) 875-2622. Fax: (604) 875-3777. E-mail: ethomas{at}cw.bc.ca.

Journal of Clinical Microbiology, April 2004, p. 1409-1413, Vol. 42, No. 4
0095-1137/04/$08.00+0     DOI: 10.1128/JCM.42.4.1409-1413.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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Antimicrob. Agents Chemother. Clin. Microbiol. Rev.
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Copyright © 2004 by the American Society for Microbiology. All rights reserved.