Detection and genotyping of varicella-zoster virus by TaqMan allelic discrimination real-time PCR.
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.
Campsall
Paul A
PA
Department of Pathology and Laboratory Medicine, University of British Columbia and Children's & Women's Health Centre of British Columbia, Canada.
Au
Nicholas H C
NH
Prendiville
Julie S
JS
Speert
David P
DP
Tan
Rusung
R
Thomas
Eva E
EE
D000483:Alleles; D002644:Chickenpox / Q000821:virology; D019433:Chickenpox Vaccine; D015342:DNA Probes; D004279:DNA, Viral / Q000032:analysis; D005260:Female; D005838:Genotype; D006562:Herpes Zoster / Q000821:virology; D014645:Herpesvirus 3, Human / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification*; D006801:Humans; D007223:Infant; D016133:Polymerase Chain Reaction / Q000379:methods*; D012150:Polymorphism, Restriction Fragment Length; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D019914:Taq Polymerase / Q000378:metabolism*
D019433:Chickenpox Vaccine; D015342:DNA Probes; D004279:DNA, Viral; D019914:Taq Polymerase