Discriminating between varicella-zoster virus vaccine and wild-type strains by loop-mediated isothermal amplification

Yuki Higashimoto, Masaru Ihira, Akane Ohta, Shigeki Inoue, Chie Usui, Yoshizo Asano, Tetsushi Yoshikawa

Research output: Contribution to journalArticle

22 Citations (Scopus)

Abstract

The loop-mediated isothermal amplification (LAMP) method was developed to distinguish between the varicella-zoster virus (VZV) vaccine (vOka) strain and wild-type strains. Two single nucleotide polymorphisms (SNPs) (nucleotide [nt] 105705 for VR-1 VZV LAMP and nt 106262 for VR-2 VZV LAMP) located in the open reading frame 62 gene were selected as LAMP targets. Amplified vOka DNA demonstrated a typical ladder pattern; however, no LAMP product was detected in reactions performed with DNAs from other human herpesviruses by either VR-1 VZV LAMP or VR-2 VZV LAMP. This result was confirmed by a turbidity assay. The sensitivities of both VR-1 and VR-2 VZV LAMP determined by either the turbidity assay or agarose gel electrophoresis were 100 copies per reaction. To discriminate the vOka strain from wild-type strains, VR-1 and VR-2 VZV LAMP products were digested with the appropriate restriction enzymes (SacII for VR-1 LAMP and SmaI for VR-2 LAMP). The digested products were clearly different in the vOka strain and wild-type strains. To evaluate the utility of the LAMP methods for rapid differentiation, viral DNA (without DNA extraction) in swab samples was directly tested. Wild-type VZV DNA was detected in 20 swab samples by either VR-1 VZV LAMP or VR-2 VZV LAMP. Sequence analysis confirmed the expected SNPs in the LAMP products amplified from the vOka strain and the five wild-type strains.

Original languageEnglish
Pages (from-to)2665-2670
Number of pages6
JournalJournal of clinical microbiology
Volume46
Issue number8
DOIs
Publication statusPublished - 08-2008

All Science Journal Classification (ASJC) codes

  • Microbiology (medical)

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