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.
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.
Harbecke
Ruth
R
Department of Veterans Affairs San Diego Healthcare System, San Diego, California, USA.
Oxman
Michael N
MN
Arnold
Beth A
BA
Ip
Charlotte
C
Johnson
Gary R
GR
Levin
Myron J
MJ
Gelb
Lawrence D
LD
Schmader
Kenneth E
KE
Straus
Stephen E
SE
Wang
Hui
H
Wright
Peter F
PF
Pachucki
Constance T
CT
Gershon
Anne A
AA
Arbeit
Robert D
RD
Davis
Larry E
LE
Simberkoff
Michael S
MS
Weinberg
Adriana
A
Williams
Heather M
HM
Cheney
Carol
C
Petrukhin
Luba
L
Abraham
Katalin G
KG
Shaw
Alan
A
Manoff
Susan
S
Antonello
Joseph M
JM
Green
Tina
T
Wang
Yue
Y
Tan
Charles
C
Keller
Paul M
PM
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
D019433:Chickenpox Vaccine; D017931:DNA Primers; D022301:Herpes Simplex Virus Vaccines; D014612:Vaccines; D055544:beta-Globins