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Appl Environ Microbiol. Jan 1994; 60(1): 149–152.
PMCID: PMC201282

Detection of mycoplasma contamination in cell cultures by a mycoplasma group-specific PCR.

Abstract

The suitability of a 16S rRNA-based mycoplasma group-specific PCR for the detection of mycoplasma contamination in cell cultures was investigated. A total of 104 cell cultures were tested by using microbiological culture, DNA fluorochrome staining, DNA-rRNA hybridization, and PCR techniques. A comparison of the results obtained with these techniques revealed agreement for 95 cell cultures. Discrepant results, which were interpreted as false negative or false positive on the basis of a comparison with the results obtained with other methods, were observed with nine cell cultures. The microbiological culture technique produced false-negative results for four cell cultures. The hybridization technique produced false-negative results for two cell cultures, and for one of these cell cultures the DNA staining technique also produced a false-negative result. The PCR may have produced false-positive results for one cell culture. Ambiguous results were obtained with the remaining two cell cultures. Furthermore, the presence of contaminating bacteria interfered with the interpretation of the DNA staining results for 16 cell cultures. For the same reason the hybridization signals of nine cell cultures could not be interpreted. Our results demonstrate the drawbacks of each of the detection methods and the suitability of the PCR for the detection of mycoplasmas in cell cultures.

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Selected References

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  • Göbel UB, Stanbridge EJ. Cloned mycoplasma ribosomal RNA genes for the detection of mycoplasma contamination in tissue cultures. Science. 1984 Dec 7;226(4679):1211–1213. [PubMed]
  • Hay RJ, Macy ML, Chen TR. Mycoplasma infection of cultured cells. Nature. 1989 Jun 8;339(6224):487–488. [PubMed]
  • Johansson KE, Johansson I, Göbel UB. Evaluation of different hybridization procedures for the detection of mycoplasma contamination in cell cultures. Mol Cell Probes. 1990 Feb;4(1):33–42. [PubMed]
  • Kwok S, Higuchi R. Avoiding false positives with PCR. Nature. 1989 May 18;339(6221):237–238. [PubMed]
  • Mattsson JG, Johansson KE. Oligonucleotide probes complementary to 16S rRNA for rapid detection of mycoplasma contamination in cell cultures. FEMS Microbiol Lett. 1993 Mar 1;107(2-3):139–144. [PubMed]
  • Razin S, Gross M, Wormser M, Pollack Y, Glaser G. Detection of mycoplasmas infecting cell cultures by DNA hybridization. In Vitro. 1984 May;20(5):404–408. [PubMed]
  • Spaepen M, Angulo AF, Marynen P, Cassiman JJ. Detection of bacterial and mycoplasma contamination in cell cultures by polymerase chain reaction. FEMS Microbiol Lett. 1992 Nov 15;78(1):89–94. [PubMed]
  • Stanbridge E. Mycoplasmas and cell cultures. Bacteriol Rev. 1971 Jun;35(2):206–227. [PMC free article] [PubMed]
  • van Kuppeveld FJ, van der Logt JT, Angulo AF, van Zoest MJ, Quint WG, Niesters HG, Galama JM, Melchers WJ. Genus- and species-specific identification of mycoplasmas by 16S rRNA amplification. Appl Environ Microbiol. 1992 Aug;58(8):2606–2615. [PMC free article] [PubMed]

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