迅速・簡易遺伝子診断法の開発 [in Japanese] Development of rapid and simple genomic diagnostic method [in Japanese]
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To develop the rapid and simple genomic diagnostic method, we analyzed the partial <i>dna</i>A sequence of 27 mycobacterial species. The partial <i>dna</i>A sequence could distinguish <i>M. kansasii</i> and <i>M. gastri</i>. Based on this region and RLEP sequence of <i>M. leprae</i>, we established the loop-mediated isothermal amplification method (LAMP) to detect each species. The LAMP method for <i>M. kansasii</i> and <i>M. gastri</i>, could detect 500 copies. Five copies of <i>M. leprae</i> genomic DNA could be detect in 30min. To simplify the sample processing, the LAMP assay was performed with FTA filter paper. <i>M. leprae</i> bacilli were applied on filter paper that lyses bacilli and bound DNA, eliminating sample centrifugation and extraction procedures. Assays of number standards showed reproducible detection rate 50 bacilli of <i>M. leprae</i>. Thus, The LAMP assay combined with FTA card has the advantages of rapid and simple detection and provides a practical, economical, and specific method for the diagnosis of <i>M. leprae</i> and NTM infection.
- Japanese journal of leprosy
Japanese journal of leprosy 75(3), 265-269, 2006-10-01
Japanese Leprosy Association