Detection of mycoplasmas infecting cell cultures by DNA hybridization

Summary

Infection of cell cultures by mycoplasmas can be detected and the mycoplasma identified by Southern blot hybridization of theEco RI-digested DNA of the suspected cell cultures with a nick-translated probe consisting of cloned ribosomal RNA genes ofMycoplasma capricolum. The probe does not hybridize with eukaryotic DNA. The hybridization pattern with mycoplasmal DNA is species specific, enabling the identification of the four most prevalent mycoplasma contaminants,Mycoplasma orale, Mycoplasma hyorhinis, Mycoplasma arginini, andAcholeplasma laidlawii. The test is also very sensitive and can detect as little as 1 ng of mycoplasmal DNA, roughly equivalent to the DNA content of 105 mycoplasmas.

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The study was supported by the U.S. Public Health Service Grant GM25286 awarded to G. G. and by a grant from the United States-Israel Binational Agricultural Research Development Fund (BARD) awarded to S. R.

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Razin, S., Gross, M., Wormser, M. et al. Detection of mycoplasmas infecting cell cultures by DNA hybridization. In Vitro 20, 404–408 (1984). https://doi.org/10.1007/BF02619586

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Key words

  • mycoplasma infection
  • cell culture infection
  • DNA hybridization