An Enrichment Medium for Increasing a Very Small Number of <i>Vibrio parahaemolyticus</i> Cells to the Detection Limit of the Loop-Mediated Isothermal Amplification (LAMP) Assay

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  • Yamazaki Mitsugu
    Laboratory of Microbiology, Aichi Prefectural Kinuura-Tobu Health Center, Japan Department of Microbiology and Medical Zoology, Aichi Prefectural Institute of Public Health, Japan
  • Aoki Hidemi
    Laboratory of Microbiology, Aichi Prefectural Kinuura-Tobu Health Center, Japan Department of Microbiology and Medical Zoology, Aichi Prefectural Institute of Public Health, Japan
  • Iwade Yoshito
    Microbiological Research Section, Mie Prefectural Health and Environment Research Institute, Japan
  • Matsumoto Masakado
    Department of Microbiology and Medical Zoology, Aichi Prefectural Institute of Public Health, Japan
  • Yamada Kazuhiro
    Department of Microbiology and Medical Zoology, Aichi Prefectural Institute of Public Health, Japan
  • Yamamoto Hiroaki
    Department of Microbiology and Medical Zoology, Aichi Prefectural Institute of Public Health, Japan
  • Suzuki Masahiro
    Department of Microbiology and Medical Zoology, Aichi Prefectural Institute of Public Health, Japan
  • Hiramatsu Reiji
    Department of Microbiology and Medical Zoology, Aichi Prefectural Institute of Public Health, Japan
  • Minagawa Hiroko
    Department of Microbiology and Medical Zoology, Aichi Prefectural Institute of Public Health, Japan

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  • An Enrichment Medium for Increasing a Very Small Number of Vibrio parahaemolyticus Cells to the Detection Limit of the Loop-Mediated Isothermal Amplification (LAMP) Assay

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Abstract

<p>We developed an enrichment medium for use with the loop-mediated isothermal amplification (LAMP) assay (enrichment media + LAMP assay) to quickly increase a very small number of<tt> </tt>Vibrio parahaemolyticus<tt> </tt>cells to the detection limit of the assay. Thirty-nine different enrichment media were prepared based on evaluating 12 potential ingredients. From our assessment of the 39 media, enrichment medium #36, which contained 2% sodium chloride, 1% proteose peptone no. 2, 0.1% trehalose, 0.5% α-ketoglutaric acid, 0.25% pyruvic acid, and 0.5% yeast extract (pH 8.6), was found to be most effective at enhancing the proliferation of<tt> </tt>V. parahaemolyticus<tt> </tt>during incubation for 3 h at 40°C. We compared the detection limits of the LAMP assay, the enrichment medium #36 + LAMP assay, and the cultivation method using bacterial cell and spiked shrimp sample tests. The detection limits of the LAMP assay, the medium #36 + LAMP assay, and the cultivation method were 103, 100–10-1, and 10-1<tt> </tt>CFU ml-1, respectively. Enrichment medium #36 promoted a 103- to 104-fold increase in the bacterial population, and the detection limit of the enrichment media + LAMP assay was the same as that of the cultivation method.</p>

Journal

  • Japanese Journal of Infectious Diseases

    Japanese Journal of Infectious Diseases 65 (2), 111-116, 2012-03-30

    National Institute of Infectious Diseases, Japanese Journal of Infectious Diseases Editorial Committee

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