魚介類中の腸炎ビブリオ定量のためのMPN-PCR法の評価 Evaluation of MPN Method Combined with PCR Procedure for Detection and Enumeration of Vibrio parahaemolyticus in Seafood

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抄録

腸炎ビブリオと <i>Vibrio alginolyticus</i> を接種した魚介類および自然汚染魚介類について,易熱性ヘモリシン遺伝子(<i>tlh</i>)を標的としたMPN-PCR法とアルカリペプトン水(APW)増菌およびTCBS培養の組合せによるMPN法(MPN-TCBS法) により腸炎ビブリオ菌数を測定した.MPN-TCBS法ではTCBS培地上に発育した<i>V. alginolyticus</i> などの影響により腸炎ビブリオの分離が困難となる例が見られた.一方MPN-PCR法は,腸炎ビブリオを分離する必要なくAPWから<i>tlh</i> 遺伝子を検出することが可能であったことから,魚介類,特に腸炎ビブリオ以外の菌が多数TCBS培地上に発育する検体では,MPN-TCBS法により腸炎ビブリオの菌数測定に適していると考えられた.

<i>Vibrio parahaemolyticus</i> densities in spiked and naturally contaminated seafood samples were enumerated by the MPN method combined with a PCR procedure (MPN-PCR method) targeting the species-specific thermolabile hemolysin gene (<i>tlh</i>), and by the MPN method using subcultivation of alkaline-peptone-water (APW) enrichment culture on thiosulfate-citrate-bile-sucrose (TCBS) agar (MPN-TCBS method). In the samples spiked with both <i>V. parahaemolyticus</i> and <i>V. alginolyticus</i>, the numbers of <i>V. parahaemolyticus</i> enumerated by the MPN-PCR method were similar to, or higher than the numbers of spiked cells, whereas those enumerated by the MPN-TCBS method were below the numbers of spiked cells. In naturally contaminated seafood samples, the numbers of <i>V. parahaemolyticus</i> enumerated by the MPN-PCR method were higher than those by the MPN-TCBS method. In the case of the MPN-TCBS method, isolation of <i>V. parahaemolyticus</i> from some APW cultures was difficult because of the overgrowth of many colonies other than <i>V. parahaemolyticus</i> (<i>e.g</i>., <i>V. alginolyticus</i>) on TCBS agar. In contrast, the PCR technique could detect <i>tlh</i> from APW culture without isolation of <i>V. parahaemolyticus</i>, so the possibility of failing to obtain a positive result in APW culture by the MPN-PCR method was considered to be lower than that by the MPN-TCBS method. Furthermore, utilization of the PCR technique reduces the time and labor required for the biochemical identification tests used in the MPN-TCBS method. For the detection and enumeration of <i>V. parahaemolyticus</i> in seafood, especially for samples that show many colonies other than <i>V. parahaemolyticus</i> on TCBS agar, the MPN-PCR method may be more convenient and reliable than the MPN-TCBS method.

収録刊行物

  • 食品衛生学雑誌 = Journal of the Food Hygienics Society of Japan

    食品衛生学雑誌 = Journal of the Food Hygienics Society of Japan 44(6), 289-293, 2003-12-25

    公益社団法人 日本食品衛生学会

参考文献:  11件中 1-11件 を表示

被引用文献:  2件中 1-2件 を表示

各種コード

  • NII論文ID(NAID)
    10013952449
  • NII書誌ID(NCID)
    AN00117741
  • 本文言語コード
    ENG
  • 資料種別
    ART
  • ISSN
    00156426
  • NDL 記事登録ID
    6812588
  • NDL 雑誌分類
    ZS17(科学技術--医学--衛生学・公衆衛生)
  • NDL 請求記号
    Z19-364
  • データ提供元
    CJP書誌  CJP引用  NDL  J-STAGE 
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