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.
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.
authors.affiliation: Department of Veterans Affairs San Diego Healthcare System, San Diego, California, USA.
authors.forename: Ruth
authors.initials: R
authors.lastname: Harbecke
authors.forename: Michael N
authors.initials: MN
authors.lastname: Oxman
authors.forename: Beth A
authors.initials: BA
authors.lastname: Arnold
authors.forename: Charlotte
authors.initials: C
authors.lastname: Ip
authors.forename: Gary R
authors.initials: GR
authors.lastname: Johnson
authors.forename: Myron J
authors.initials: MJ
authors.lastname: Levin
authors.forename: Lawrence D
authors.initials: LD
authors.lastname: Gelb
authors.forename: Kenneth E
authors.initials: KE
authors.lastname: Schmader
authors.forename: Stephen E
authors.initials: SE
authors.lastname: Straus
authors.forename: Hui
authors.initials: H
authors.lastname: Wang
authors.forename: Peter F
authors.initials: PF
authors.lastname: Wright
authors.forename: Constance T
authors.initials: CT
authors.lastname: Pachucki
authors.forename: Anne A
authors.initials: AA
authors.lastname: Gershon
authors.forename: Robert D
authors.initials: RD
authors.lastname: Arbeit
authors.forename: Larry E
authors.initials: LE
authors.lastname: Davis
authors.forename: Michael S
authors.initials: MS
authors.lastname: Simberkoff
authors.forename: Adriana
authors.initials: A
authors.lastname: Weinberg
authors.forename: Heather M
authors.initials: HM
authors.lastname: Williams
authors.forename: Carol
authors.initials: C
authors.lastname: Cheney
authors.forename: Luba
authors.initials: L
authors.lastname: Petrukhin
authors.forename: Katalin G
authors.initials: KG
authors.lastname: Abraham
authors.forename: Alan
authors.initials: A
authors.lastname: Shaw
authors.forename: Susan
authors.initials: S
authors.lastname: Manoff
authors.forename: Joseph M
authors.initials: JM
authors.lastname: Antonello
authors.forename: Tina
authors.initials: T
authors.lastname: Green
authors.forename: Yue
authors.initials: Y
authors.lastname: Wang
authors.forename: Charles
authors.initials: C
authors.lastname: Tan
authors.forename: Paul M
authors.initials: PM
authors.lastname: Keller
mesh_terms: D019433:Chickenpox Vaccine; D017931:DNA Primers; D003937:Diagnosis, Differential; D006561:Herpes Simplex / Q000175:diagnosis; D022301:Herpes Simplex Virus Vaccines; D006562:Herpes Zoster / Q000175:diagnosis; D014645:Herpesvirus 3, Human / Q000145:classification* / Q000235:genetics* / Q000302:isolation & purification; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods* / Q000592:standards; D020641:Polymorphism, Single Nucleotide; D012015:Reference Standards; D012680:Sensitivity and Specificity; D018139:Simplexvirus / Q000145:classification* / Q000235:genetics* / Q000302:isolation & purification; D014612:Vaccines; D055544:beta-Globins / Q000235:genetics
chemical_list: D019433:Chickenpox Vaccine; D017931:DNA Primers; D022301:Herpes Simplex Virus Vaccines; D014612:Vaccines; D055544:beta-Globins