1. A real-time PCR assay to identify and discriminate among wild-type and vaccine strains of varicella-zoster virus and herpes simplex virus in clinical specimens, and comparison with the clinical diagnoses.
- Author
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Harbecke R, Oxman MN, Arnold BA, Ip C, Johnson GR, Levin MJ, Gelb LD, Schmader KE, Straus SE, Wang H, Wright PF, Pachucki CT, Gershon AA, Arbeit RD, Davis LE, Simberkoff MS, Weinberg A, Williams HM, Cheney C, Petrukhin L, Abraham KG, Shaw A, Manoff S, Antonello JM, Green T, Wang Y, Tan C, and Keller PM
- Subjects
- DNA Primers, Diagnosis, Differential, Herpes Simplex diagnosis, Herpes Zoster diagnosis, Herpesvirus 3, Human isolation & purification, Humans, Polymerase Chain Reaction standards, Polymorphism, Single Nucleotide, Reference Standards, Sensitivity and Specificity, Simplexvirus isolation & purification, Vaccines, beta-Globins genetics, Chickenpox Vaccine, Herpes Simplex Virus Vaccines, Herpesvirus 3, Human classification, Herpesvirus 3, Human genetics, Polymerase Chain Reaction methods, Simplexvirus classification, Simplexvirus genetics
- Abstract
A real-time PCR assay was developed to identify varicella-zoster virus (VZV) and herpes simplex virus (HSV) DNA in clinical specimens from subjects with suspected herpes zoster (HZ; shingles). Three sets of primers and probes were used in separate PCR reactions to detect and discriminate among wild-type VZV (VZV-WT), Oka vaccine strain VZV (VZV-Oka), and HSV DNA, and the reaction for each virus DNA was multiplexed with primers and probe specific for the human beta-globin gene to assess specimen adequacy. Discrimination of all VZV-WT strains, including Japanese isolates and the Oka parent strain, from VZV-Oka was based upon a single nucleotide polymorphism at position 106262 in ORF 62, resulting in preferential amplification by the homologous primer pair. The assay was highly sensitive and specific for the target virus DNA, and no cross-reactions were detected with any other infectious agent. With the PCR assay as the gold standard, the sensitivity of virus culture was 53% for VZV and 77% for HSV. There was 92% agreement between the clinical diagnosis of HZ by the Clinical Evaluation Committee and the PCR assay results., (Published 2009 Wiley-Liss, Inc.)
- Published
- 2009
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