In situ PCR法による腸管出血性大腸菌の検出

  title={In situ PCR法による腸管出血性大腸菌の検出},
  author={黒川 顕 and 谷 佳津治 and 那須 正夫},
  journal={Japanese journal of bacteriology},
Direct in situ PCR法は特定のDNA配列を標的として菌体内でPCRを行い, 増幅されたDNAを蛍光基質などでラベルすることにより, 蛍光顕微鏡下で特定遺伝子を持つ細菌を単個細胞レベルで特異的に検出する方法である。PCRには腸管出血性大腸菌の Vero 毒素をコードする遺伝子 slt-I, slt-II に特異的なEVT, EVSプライマーを用いた。増幅されたDNAはジゴキシゲニンにより標識し, アルカリホスファターゼ標識した抗ジゴキシゲニン抗体および蛍光基質としてHNPP/Fast Red TRを用いた。その結果, E. coli O157を蛍光顕微鏡下で特異的にまた高感度に単個細胞レベルで検出することができた。 
8 Citations
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