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Reverse transcription-polymerase chain reaction detection and sequence analysis of small round-structured viruses in Japan

  • K. Yamazaki
  • M. Oseto
  • Y. Seto
  • E. Utagawa
  • T. Kimoto
  • Y. Minekawa
  • S. Inouye
  • S. Yamazaki
  • Y. Okuno
  • I. Oishi
Conference paper
Part of the Archives of Virology book series (ARCHIVES SUPPL, volume 12)

Summary

Between 1985 and 1995, mass outbreaks of acute gastroenteritis caused by small round-structured virus (SRSV), occurred in eight prefectures in Japan. Fecal samples from 59 patients ill during these outbreaks were recently examined in our laboratory by electron microscopy (EM) and by reverse transcription-polymerase chain reaction (RT-PCR). For RT-PCR, we prepared two sets of primers, a set corresponding to the polymerase region of open reading frame 1 (ORF-1) and a set corresponding to the capsid region of ORF-2 of Norwalk virus (NV). The SRSV nucleic acid detection rate with these primers was more than double that achieved with EM. Most samples found by EM to contain virus particles were also positive by PCR. When the two sets of primers were used separately, the virus detection rate differed depending on the primer used, suggesting that the viral strains examined were not genetically not homogeneous. We then selected nine strains of the virus, cloned their PCR products and analyzed their base sequences. The base sequences of these strains were compared with those of reference strains including prototype NV and Snow Mountain agent (SMA). This comparison yielded the following findings: (1) SRSVs that cause mass outbreaks of gastroenteritis in Japan are genetically variable; (2) SRSV strains that are genetically similar to SMA and SRSV-OTH25/89/J (OTH25) are dominant in Japan, but strains similar to NV are also present in this country; and (3) a strain (MI1/94) which is genetically identical to Southampton virus (SHV) was detected. Detection of SRSV using sensitive RT-PCR and analysis of the sequences of the amplification products seems to provide a useful means of studying the molecular epidemiology of SRSV.

Keywords

Fecal Sample Base Sequence Acute Gastroenteritis Viral Nucleic Acid Norwalk Virus 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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Copyright information

© Springer-Verlag Wien 1996

Authors and Affiliations

  • K. Yamazaki
    • 1
  • M. Oseto
    • 2
  • Y. Seto
    • 3
  • E. Utagawa
    • 4
  • T. Kimoto
    • 1
  • Y. Minekawa
    • 1
  • S. Inouye
    • 4
  • S. Yamazaki
    • 4
  • Y. Okuno
    • 1
  • I. Oishi
    • 1
  1. 1.Laboratory of VirologyOsaka Prefectural Institute of Public HealthOsaka 537Japan
  2. 2.Ehime Prefectural Institute of Public HealthEhimeJapan
  3. 3.Osaka Municipal Institute of Environment and ScienceOsakaJapan
  4. 4.National Institute of HealthTokyoJapan

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