1. Detection and genotyping of varicella-zoster virus by TaqMan allelic discrimination real-time PCR.
- Author
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Campsall PA, Au NH, Prendiville JS, Speert DP, Tan R, and Thomas EE
- Subjects
- Chickenpox virology, Chickenpox Vaccine, DNA Probes, DNA, Viral analysis, Female, Genotype, Herpes Zoster virology, Herpesvirus 3, Human genetics, Humans, Infant, Polymorphism, Restriction Fragment Length, Reproducibility of Results, Sensitivity and Specificity, Alleles, Herpesvirus 3, Human classification, Herpesvirus 3, Human isolation & purification, Polymerase Chain Reaction methods, Taq Polymerase metabolism
- Abstract
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.
- Published
- 2004
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