Cycling probe technology to quantify and discriminate between wild-type varicella-zoster virus and Oka vaccine strains

Masaru Ihira, Yuki Higashimoto, Yoshiki Kawamura, Ken Sugata, Masahiro Ohashi, Yoshizo Asano, Tetsushi Yoshikawa

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

Rapid differentiation between wild-type varicella zoster virus (VZV) and Oka-vaccine (vOka) strains is important for monitoring side reactions of varicella vaccination. To develop a high-throughput molecular diagnostic method for the differentiation of wild-type VZV and vOka strains based on cycling probe technology. The primers were designed to amplify common sequences spanning a single nucleotide polymorphism (SNP) in gene 62 of VZV. DNA-RNA chimeric probes (cycling probes) were designed to detect the SNP at nucleotide 105705. The cycling probe real-time PCR assays for VZV wild-type and vOka strains specifically amplified plasmids containing target sequences that ranged between 10 and 1×106 copies per reaction. The inter- and intra-assay coefficients of variation were less than 5%. After initial validation studies, the clinical reliability of this method was evaluated using 38 swab samples that were collected from patients suspected of being zoster. Compared to the loop mediated isothermal amplification method, which is defined as the gold standard, cycling probe real-time PCR was highly sensitive and specific. The cycling probe real-time PCR technology is a reliable tool for differentiating between wild-type VZV and vOka strains in clinical samples.

Original languageEnglish
Pages (from-to)308-313
Number of pages6
JournalJournal of Virological Methods
Volume193
Issue number2
DOIs
Publication statusPublished - 01-11-2013

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Human Herpesvirus 3
Vaccines
Technology
Real-Time Polymerase Chain Reaction
Single Nucleotide Polymorphism
RNA Probes
Molecular Pathology
Chickenpox
Validation Studies
Herpes Zoster
Vaccination
Plasmids
Nucleotides
DNA
Genes

All Science Journal Classification (ASJC) codes

  • Virology

Cite this

Ihira, Masaru ; Higashimoto, Yuki ; Kawamura, Yoshiki ; Sugata, Ken ; Ohashi, Masahiro ; Asano, Yoshizo ; Yoshikawa, Tetsushi. / Cycling probe technology to quantify and discriminate between wild-type varicella-zoster virus and Oka vaccine strains. In: Journal of Virological Methods. 2013 ; Vol. 193, No. 2. pp. 308-313.
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Cycling probe technology to quantify and discriminate between wild-type varicella-zoster virus and Oka vaccine strains. / Ihira, Masaru; Higashimoto, Yuki; Kawamura, Yoshiki; Sugata, Ken; Ohashi, Masahiro; Asano, Yoshizo; Yoshikawa, Tetsushi.

In: Journal of Virological Methods, Vol. 193, No. 2, 01.11.2013, p. 308-313.

Research output: Contribution to journalArticle

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