A Fluorogenic Assay for the Rapid Detection of Some <i>Vibrio</i> Species Including <i>Vibrio parahaemolyticus</i> in Foods

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  • MIYAMOTO Takahisa
    Department of Food Science and Technology, Faculty of Agriculture, Kyushu University
  • SHEU Yea-Ing
    Department of Food Science and Technology, Faculty of Agriculture, Kyushu University
  • MIWA Harufumi
    Ajinomoto Co., Inc.
  • HATANO Shoji
    Department of Food Science and Technology, Faculty of Agriculture, Kyushu University

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  • 蛍光法による腸炎ビブリオを含む2, 3のビブリオ属細菌の迅速検査法
  • ケイコウホウ ニ ヨル チョウエンビブリオ オ フクム 2 3 ノ ビブリオゾ

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Abstract

A rapid assay method for some Vibrio species including Vibrio parahaemolyticus was developed. The assay involved the enrichment culture of Vibrio species in BSP-HP medium (0.2% bacto-tryptone, 2% sodium chloride, 250 units/ml polymyxin B sulfate, 0.025% sodium hexametaphosphate, pH 8.5) and the specific measurement of intracellular trypsin-like activity by using the fluorogenic substrate benzoyl-L-arginine-7-aminomethylcoumarin. In BSP-HP medium, growth and trypsin-like activity of bacteria other than V. parahaemolyticus, V. alginolyticus and V. harveyi were suppressed. Although trypsin-like enzyme in food samples interfered with the fluorogenic assay, the interference could be overcome by the addition of 1mM EDTA to the reaction mixture. Using this fluorogenic assay with commercial seafoods, 400 cells of V. parahaemolyticus per gram of food sample were detected within 8hr. Trypsinlike activity measured by the assay was proportional to the number of V. parahaemolyticus determined by the conventional BTB teepol agar plating method (correlation coefficient r=0.97). In the medium, V. alginolyticus, one of the V. parahaemolyticus-allied bacteria, and V. harveyi also showed vigorous growth and high trypsin-like activity. A more specific culture condition for V. parahaemolyticus is required, and further investigation is in progress.

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