Rapid detection of varicella-zoster virus infection by a loop-mediated isothermal amplification method

Souki Okamoto, Tetsushi Yoshikawa, Masaru Ihira, Kayoko Suzuki, Kaoru Shimokata, Yukihiro Nishiyama, Yoshizo Asano

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Abstract

The reliability of varicella-zoster virus (VZV) loop-mediated isothermal amplification (LAMP) was evaluated for rapid diagnosis of viral infection. VZV-specific primers only amplified VZV DNA; no LAMP products were observed in reactions performed with other viral DNA templates. The specificity of this method was confirmed by two independent determinations, agarose gel electrophoresis and a turbidity assay. The sensitivity of VZV LAMP, determined by agarose gel electrophoresis, were 500 copies/tube. Detection using the turbidity assay, however, gave a sensitivity of 1,000 copies/tube. After these initial validation studies, reliability of VZV LAMP was evaluated for the detection of viral DNA in clinical specimens. Thirty-two swab samples collected from patients with vesicular skin eruptions were tested for VZV DNA. VZV was confirmed in sample numbers 10-32 by VZV real-time PCR, a previously established technique. VZV LAMP products were detected using turbidity from samples 13 to 32 (sensitivity; 87.0%, specificity; 100%, positive predictive value; 100%, negative predictive value; 75%). Although low levels of VZV DNA could be detected in the three samples exhibiting divergent results (samples numbers 10-12), no VZV LAMP product was detected in these samples, indicating a higher detection limit for this assay. Requirement of a DNA extraction step in the VZV LAMP method was examined in next experiment. The turbidity assay detected a VZV LAMP product in all of the 20 positive swab samples (samples numbers 13-32), regardless of DNA extraction.

Original languageEnglish
Pages (from-to)677-682
Number of pages6
JournalJournal of Medical Virology
Volume74
Issue number4
DOIs
Publication statusPublished - 01-12-2004

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Human Herpesvirus 3
Virus Diseases
DNA
Agar Gel Electrophoresis
Viral DNA
Validation Studies
Limit of Detection
Real-Time Polymerase Chain Reaction

All Science Journal Classification (ASJC) codes

  • Virology
  • Infectious Diseases

Cite this

Okamoto, Souki ; Yoshikawa, Tetsushi ; Ihira, Masaru ; Suzuki, Kayoko ; Shimokata, Kaoru ; Nishiyama, Yukihiro ; Asano, Yoshizo. / Rapid detection of varicella-zoster virus infection by a loop-mediated isothermal amplification method. In: Journal of Medical Virology. 2004 ; Vol. 74, No. 4. pp. 677-682.
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Rapid detection of varicella-zoster virus infection by a loop-mediated isothermal amplification method. / Okamoto, Souki; Yoshikawa, Tetsushi; Ihira, Masaru; Suzuki, Kayoko; Shimokata, Kaoru; Nishiyama, Yukihiro; Asano, Yoshizo.

In: Journal of Medical Virology, Vol. 74, No. 4, 01.12.2004, p. 677-682.

Research output: Contribution to journalArticle

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AU - Okamoto, Souki

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