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.
Title | 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. |
Publication Type | Journal Article |
Year of Publication | 2009 |
Authors | 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, Keller PM |
Corporate Authors | Shingles Prevention Study Group |
Journal | J Med Virol |
Volume | 81 |
Issue | 7 |
Pagination | 1310-22 |
Date Published | 2009 Jul |
ISSN | 1096-9071 |
Keywords | beta-Globins, Chickenpox Vaccine, Diagnosis, Differential, DNA Primers, Herpes Simplex, Herpes Simplex Virus Vaccines, Herpes Zoster, Herpesvirus 3, Human, Humans, Polymerase Chain Reaction, Polymorphism, Single Nucleotide, Reference Standards, Sensitivity and Specificity, Simplexvirus, Vaccines |
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. |
DOI | 10.1002/jmv.21506 |
Alternate Journal | J. Med. Virol. |
PubMed ID | 19475609 |
PubMed Central ID | PMC4217208 |
Grant List | Z01 AI000856-08 / / Intramural NIH HHS / United States |