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

• Published in: Journal of medical virology Vol. 81; no. 7; pp. 1310 - 1322
• Main Authors: Harbecke, Ruth, Oxman, Michael N, Arnold, Beth A, Ip, Charlotte, Johnson, Gary R, Levin, Myron J, Gelb, Lawrence D, Schmader, Kenneth E, Straus, Stephen E, Wang, Hui, Wright, Peter F, Pachucki, Constance T, Gershon, Anne A, Arbeit, Robert D, Davis, Larry E, Simberkoff, Michael S, Weinberg, Adriana, Williams, Heather M, Cheney, Carol, Petrukhin, Luba, Abraham, Katalin G, Shaw, Alan, Manoff, Susan, Antonello, Joseph M, Green, Tina, Wang, Yue, Tan, Charles, Keller, Paul M
• Format: Journal Article
• Language: English
• Published: Hoboken Wiley Subscription Services, Inc., A Wiley Company 01.07.2009; Wiley
• Subjects: beta-Globins - genetics; Biological and medical sciences; Chickenpox Vaccine; clinical trial; diagnosis of herpes zoster; Diagnosis, Differential; DNA Primers; Fundamental and applied biological sciences. Psychology; Herpes Simplex - diagnosis; Herpes Simplex Virus Vaccines; Herpes Zoster - diagnosis; Herpesvirus 3, Human - classification; Herpesvirus 3, Human - genetics; Herpesvirus 3, Human - isolation & purification; Human viral diseases; Humans; Infectious diseases; Medical sciences; Microbiology; Miscellaneous; Polymerase Chain Reaction - methods; Polymerase Chain Reaction - standards; Polymorphism, Single Nucleotide; Reference Standards; Sensitivity and Specificity; Shingles Prevention Study; Simplexvirus - classification; Simplexvirus - genetics; Simplexvirus - isolation & purification; Techniques used in virology; Vaccines; Viral diseases; Virology; VZV ORF 62; zoster vaccine; Skin disease; Alphaherpesvirinae; Identification; Prevention; Shingles Prevention Study; zoster vaccine; Clinical trial; Herpesvirus hominis; Diagnosis; Clinical isolate; VZV ORF 62; Human; Varicella zoster virus; diagnosis of herpes zoster; Nervous system diseases; Herpes zoster; Vaccine strain; Herpesviridae; Vaccine; Method; Real time; Open reading frame; Infection; Virus; Polymerase chain reaction; Viral disease; Comparative study
• ISSN: 0146-6615; 1096-9071
• DOI: 10.1002/jmv.21506

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 β-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. J. Med. Virol. 81:1310-1322, 2009. Published 2009 Wiley-Liss, Inc.