Development of a Detection Method for the Beverage Putrefactive Bacteria, Alicyclobacillus acidoterrestris, Based on gyrB Gene

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  • GOTO Keiichi
    Microbiological & Analytical Group, Food Research Laboratories, Mitsui Norin Co ., Ltd.
  • ASAHARA Mika
    Microbiological & Analytical Group, Food Research Laboratories, Mitsui Norin Co ., Ltd.
  • KASAI Hiroaki
    Marine Biotechnology Institute Co., Ltd.
  • YOKOTA Akira
    Institute of Molecular and Cellular Biosciences, The University of Tokyo

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  • ｇｙｒＢ遺伝子内部塩基配列に基づく飲料変敗細菌Ａｌｉｃｙｃｌｏｂａｃｉｌｌｕｓ　ａｃｉｄｏｔｅｒｒｅｓｔｒｉｓの鑑別法
  • gyrB イデンシ ナイブ エンキ ハイレツ ニ モトヅク インリョウ ヘンパイ サイキン Alicyclobacillus acidoterrestris ノ カンベツホウ

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Comparative gyrB gene sequence analyses performed on type/reference strains of Alicyclobacillus species and three A. acidoterrestrisstrains demonstrated that gyrBgene was very diverse among species. However, the gyrB gene sequences of A. acidoterrestris was highly conserved within species at more than 97.6% sequence similarity . Based on these results, re-gions characteristic of A. acidoterrestriswere found and primers including that region were designed in order to specifically detect A. acidoterrestris (three forward primers and two reverse primers). Using all possible combinations of the primers, PCR reactions were carried out with A. acidoterrestris-derived DNAs. The results showed that the predicted length fragments were accurately amplified in every primer set. Apart from certain exceptions there were used in the LAMP method, it was confirmed that this amplification reaction occurred only with A. acidoterrestris. These findings indicate that A. acidoterrestris can be accurately and rapidly detected using these specific primers designed here in the PCR-electrophoresis method and/or the LAMP method.

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