A Novel Method for the Purification of DNA by Capturing Nucleic Acid and Magnesium Complexes on Non-Woven Fabric Filters under Alkaline Conditions for the Gene Diagnosis of Tuberculosis by Loop-Mediated Isothermal Amplification (LAMP)

DOI Web Site Web Site
  • Fukasawa Tadashi
    Asahi-Kasei Co., Ltd., Japan
  • Oda Naozumi
    Asahi-Kasei Co., Ltd., Japan
  • Wada Yasunao
    Clinical Laboratory, The Hospital of Hyogo College of Medicine, Japan
  • Tamaru Aki
    Department of Infectious Diseases, Osaka Prefectural Institute of Public Health, Japan
  • Fukushima Yukari
    Department of Global Epidemiology, Hokkaido University Research Center for Zoonosis Control, Japan
  • Nakajima Chie
    Department of Global Epidemiology, Hokkaido University Research Center for Zoonosis Control, Japan
  • Suzuki Yasuhiko
    Department of Global Epidemiology, Hokkaido University Research Center for Zoonosis Control, Japan

この論文をさがす

抄録

<p>A novel method for purifying DNA from clinical samples based on the complex formation of DNA and magnesium ion (Mg2+) was developed for the detection of Mycobacterium tuberculosis. The formation of a DNA-Mg2+ complex under alkaline conditions was observed by analyzing electrophoretic mobility reduction of DNA on agarose gel. The DNA-Mg2+ complex increases the efficacy of DNA recovery from the sample solution on polyethylene terephthalate non-woven fabric (PNWF) filters. Among the various divalent metal cations, only Mg2+ was associated with this effect. The applicability of DNA recovered on the PNWF filter was examined for the gene amplification method; loop-mediated isothermal amplification (LAMP). DNA on the PNWF filter could be used for the amplification of specific DNA fragments without elution from the filter. Using this method, DNA was directly purified from M. tuberculosis spiked sputum and examined by LAMP assay, showing a high sensitivity in comparison to the commercially available DNA extraction kit. These results indicated that the method developed in this study is useful for rapid gene diagnosis of tuberculosis.</p>

収録刊行物

キーワード

詳細情報 詳細情報について

問題の指摘

ページトップへ