Rapid and Direct Detection of Myocbacterium tuberculosis Complex and Mycobacteria in Sputum by Advanced Method, PCR

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  • KUSUNOKI Shinji
    Shionogi Biomedical Laboratories, Shionogi Pharmaceutical Co., Ltd.
  • MURATA Yutaka
    Shionogi Biomedical Laboratories, Shionogi Pharmaceutical Co., Ltd.
  • MINAMIDE Wakio
    Shionogi Biomedical Laboratories, Shionogi Pharmaceutical Co., Ltd.
  • UCHIDA Kiyohisa
    Shionogi Biomedical Laboratories, Shionogi Pharmaceutical Co., Ltd.
  • MIURA Hiroaki
    Department of Microbiology, Gifu University School of Medicine
  • EZAKI Takayuki
    Department of Microbiology, Gifu University School of Medicine

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Other Title
  • PCR法を用いた喀痰中の結核菌群および<I>Mycobacterium</I>属菌の迅速検出
  • PCR法を用いた喀痰中の結核菌群およびMycobacterium属菌の迅速検出
  • PCRホウ オ モチイタ カクタンチュウ ノ ケッカクキングン オヨビ Myc

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Description

We developed two PCR methods, which amplify bovine tuberculous MPB70 gene and mycobacterial 16S rRNA gene, for detection of tubercle bacilli and mycobacteria in sputum, respectively.A mong 27 Mycobacterium species and 57 species of 30 genuses other than Mycobacterium, only M.tuberculosis (TB) complex, i.e., M. tuberculosis, M. bovis, M. africanum, M. microti showed DNA amplification by PCR for MPB70, and amplification of 16S rRNA gene were observed specific in Mycobacterium species. A combination of these PCR abilities were available to differentiate the TB complex and nontuberculous mycobacteria (NTM).<BR>We investigated the correlation between these methods and conventional methods with 311 sputa that were suspected mycobacteriosis. The PCR method could detect 12 cases of TB complex and 4 cases of NTM in 17 specimens, which were positive by conventional methods, but could not for one specimen. Among 294 specimens that were negative with conventional methods, the PCR method detected 13 and 8 cases of TB complex and NTM, respectively. These results were confirmed by commercial tuberculous specific DNA probe or investigation of the clinical background of the patients. On the other hand, 273 specimens showed negative result either PCR nor conventional methods. The PCR method did not detect tuberculous DNA in normal 197 sputa, which were not suspected mycobacteriosis.<BR>These results indicate that each one of these PCR methods is highly specific to TB complex or Mycobacterium species.<BR>We concluded that these PCR methods are useful and advanced methods for rapid and direct detection of tubelculosis and mycobacteriosis.

Journal

  • Kansenshogaku Zasshi

    Kansenshogaku Zasshi 66 (12), 1682-1691, 1992

    The Japanese Association for Infectious Diseases

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