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Detection of mycoplasma contaminations by the polymerase chain reaction

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Abstract

The polymerase chain reaction (PCR) has been used for the general detection ofMollicutes. 25Mycoplasma andAcholeplasma species were detected including important contaminants of cell cultures such asM. orale, M. arginini, M. hyorhinis, M. fermentans, A. laidlawii and additional human and animal mycoplasmas. PCR reactions were performed using a set of nested primers defined from conserved regions of the 16S rRNA gene. The detection limit was determined to be 1 fg mycoplasma DNA, which is equivalent to 1–2 genome copies of the 16S rRNA coding region. The identity of the amplification products was confirmed by agarose gel electrophoresis and restriction enzyme analysis. DNA from closely and distantly related micro-organisms did not give rise to specific amplification products. The method presented here offers a much more sensitive, specific and rapid assay for the detection of mycoplasmas than the existing ones.

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Authors and Affiliations

  1. Genetik von Eukaryonten, GBF-Gesellschaft für Biotechnologische Forschung mbH, D-38124, Braunschweig, Germany

    Manfred Wirth, Martina Grashoff & Hansjörg Hauser

  2. Institute for Veterinary Medicine (Robert von Ostertag Institute) Jena Branch, Federal Health Office, D-07743, Jena, Germany

    Evelyn Berthold & Horst Pfützner

  3. Institute for Medical Immunology, Medical Division (Charité), Humboldt-University of Berlin, D-10117, Berlin, Germany

    Uli Schubert

Authors
  1. Manfred Wirth
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  2. Evelyn Berthold
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  3. Martina Grashoff
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  4. Horst Pfützner
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  5. Uli Schubert
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  6. Hansjörg Hauser
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Wirth, M., Berthold, E., Grashoff, M. et al. Detection of mycoplasma contaminations by the polymerase chain reaction. Cytotechnology 16, 67–77 (1994). https://doi.org/10.1007/BF00754609

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  • DOI: https://doi.org/10.1007/BF00754609

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