免疫磁気ビーズ法を用いた糞便材料からの Helicobacter Pylori の検出方法に関する検討  [in Japanese] Trial for Isolation of Helicobacter pylori from Fecal Specimen by Immunomagnetic Beads Separation  [in Japanese]

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Abstract

免疫磁気ビーズ法を用いて糞便材料から<I>Helicobacter pylori</I>を検出するための諸条件の検討を行った. 使用する抗<I>H. pylori</I>血清の検討では抗ホルマリン死菌体家兎血清が最も良い結果を示し, ハンクス液1mlに約5×104cfuの<I>H.pylori</I>を浮遊させて, 48cfuの菌の分離培養が可能であった. また免疫磁気ビーズ法とPCR法を組み合わせることでその検出感度を10倍高めることができた. 糞便からのH.pyloriの検出には糞便を磁気ビーズで処理する前にナイロンメッシュを通過させる, 培養には選択培地を使う, PCR (Polymerase chain reaction) 法に用いるサンプルDNAは蛋白除去するなどの操作が必要であった. <I>H.pylori</I>陽性の胃潰瘍患者より採取した糞便を用いて免疫磁気ビーズ法による分離を試みた結果PCR法は陽性であったが培養は陰性であった.

In order to clarify the possibility of feco-oral transmission of <I>Helicobacter pylori</I>, we have employed an immunomagnetic beads (IMB) separation technique and tried to isolate <SUB>H. pylori</SUB> from fecal samples. Microaerophilic cultivation and PCR of H. pylori were positive when H. pylori (10<SUP>3</SUP> cfu) was suspended in Hanks' balanced salt solution (HBSS) for the IMB separation technique. In contrast, when a lower number of <I>H. Pylori</I> (10<SUP>2</SUP> cfu) was suspended in HBSS, 2nd PCR was positive but cultivation was negative. After IMB separation technique from fecal sample containing <I>H. pylori</I> (2-10<SUP>4</SUP> cfu), 20 cfu of <I>H. pylori</I> was recovered by culture. In this case, 1st PCR was negative but 2nd PCR was positive. From a fecal sample of a patient infected with <I>H. pylori</I>, H. pylori was detected only by combination of IMB separation and PCR, although H. pylori was not recovered by culture. These results indicate that IMB separation technique followed by cultivation or PCR would be useful for estimation of the presence of <I>H. pylori</I> fecal samples.

Journal

  • Kansenshogaku Zasshi

    Kansenshogaku Zasshi 71(7), 628-633, 1997-07-20

    The Japanese Association for Infectious Diseases

References:  20

Codes

  • NII Article ID (NAID)
    10008721138
  • NII NACSIS-CAT ID (NCID)
    AN00047715
  • Text Lang
    JPN
  • Article Type
    ART
  • ISSN
    03875911
  • NDL Article ID
    4268909
  • NDL Source Classification
    ZS9(科学技術--医学--病理学・微生物学・寄生虫学・感染・免疫学・血清学・アレルギー)
  • NDL Call No.
    Z19-193
  • Data Source
    CJP  NDL  J-STAGE 
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