A PCR based detection method for Mycobacteria in biofilms from a drinking water distribution system

  • chair:

    Schwartz, T. / Kalmbach, S. / Hoffmann, S. / Szewzyk, U. / Obst, U. (1998)

  • place:

    J. Microbiol. Methods 34 (1998), 2, 113-123

  • Date: 1998
  • Schwartz, T. / Kalmbach, S. / Hoffmann, S. / Szewzyk, U. / Obst, U. (1998): „A PCR based detection method for Mycobacteria in biofilms from a drinking water distribution system“. In: J. Microbiol. Methods 34 (1998), 2, 113-123

Abstract

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A molecular biological method was developed for the selective detection of mycobacteria in biofilms from drinking water systems without requiring time consuming cultivation procedures. A newly designed 16S rDNA targeted oligonucleotide primer, which is highly specific for the genus Mycobacterium, was used for the selective PCR amplification of mycobacteria 16S rDNA fragments.

The PCR products were subsequently hybridized with a second mycobacterial-specific 16S rDNA-targeted probe. This two-step procedure greatly reduced the probability of false-positive detection of organisms not affiliated to the genus Mycobacterium. The specificity and sensitivity of the method was confirmed with various target and nontarget reference strains, followed by application in native biofilms from different drinking water distribution systems.

The results of this investigation show that mycobacteria could not be detected when groundwater was used as raw water source, but were frequently found in bank-filtered drinking water biofilms. Further PCR experiments indicated that these mycobacteria did not belong to the pathogenic or certain facultative pathogenic species of this genus, but were representatives of the environmental mycobacteria.