Summary
Studies were performed to compare three methods to detect mycoplasmal infection of cell cultures. The methods included microbiological assay by inoculation into broth and onto agar with anaerobic incubation, fluorescent DNA staining by Hoechst 33258, and mycoplasmal mediated cytotoxicity by 6 methylpurine deoxyriboside (6MPDR). Fluorescent DNA staining and 6MPDR assays were performed in an indicator cell culture system. A total of 2589 cell cultures were assayed. Mycoplasmas were detected in 174, an incidence of 6.7%. Species isolated were:Acholeplasma laidlawii, Mycoplasma orale, M. arginini, M. hyorhinis, M. fermentans, M. pirum, and M. pneumoniae. In separate studies, 6MPDR also detected infection withSpiroplasma mirum when this organisms was deliberately inoculated into cell cultures. The efficiencies of microbiological testing, fluorescent DNA assays, and 6MPDR were 43.1, 98. 8, and 97.1%, respectively.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Barile, M. F.; Hopps, H. E.; Grabowski, M. W. Incidence and sources of mycoplasma contamination: a brief review. In: McGarrity, G. J.; Murphy, D. G.; Nichols, W. W., eds. Mycoplasma infection of cell cultures. New York: Plenum Press; 1978:35–45.
Clyde, W. A., Jr.. Mycoplasma species identification based upon growth inhibition by specific antisera. J. Immunol. 92:958–965; 1964.
Del Giudice, R. A.; Gardella, R. S. Mycoplasma infection of cell cultures: effects, incidence and detection. Uses and standardization of vertebrate cell cultures. Gaithersburg, MD: Tissue Culture Association; 1985:104–115.
Del Giudice, R. A.; Tully, J. G., Rose, D. L., et al..Mycoplasma pirum, sp. nov., a terminal-structured mollicute from cell cultures. Int. J. System. Bacteriol. 35:285–291; 1985.
Göbel, U. B.; Stanbridge, E. J.. Cloned mycoplasma ribosomal RNA genes for the detection of mycoplasma contamination in tissue cultures. Science 226:1211–1213; 1984.
Kotani, H.; McGarrity, G. J.. Rapid and simple identification of mycoplasmas by immunobinding. J. Immunol. Methods 85:1–11; 1985.
McGarrity, G. J.; Coriell, L. L. Detection of anaerobic mycoplasma. In Vitro 9:17–18; 1973.
McGarrity, G. J.; Sarama, J.; Vanaman, V. Factors influencing microbiological detection of mycoplasmas in cell cultures. In Vitro 15:73–81; 1979.
McGarrity, G. J.; Carson, D. A. Adenosine phosphorylase-mediated nucleoside toxicity. Exp. Cell Res. 139:199–205. 1982.
McGarrity, G. J. Detection of mycoplasmal infection of cell cultures. In: Maramorosch, K., ed. Advances in cell culture, vol. 2 New York: Academic Press; 1982:99–131.
McGarrity, G. J.; Kotani, H.. Cell culture mycoplasmas. In: Razin, S.; Barile, M. F., eds. The mycoplasmas, vol. IV. New York: Academic Press; 1985:353–390.
McGarrity, G. J.; Sarama, J.; Vanaman, V. Cytogenetic effects of cell culture mycoplasmas: A review. In Vitro 20:1–18; 1984.
McGarrity, G. J.; Kotani, H.. Detection of cell culture mycoplasmas by a genetic probe. Exp. Cell Res. 163:273–278; 1986.
Razin, S.; Gross, M.; Wormser, M. et al. Detection of mycoplasmas infecting cell cultures by DNA hybridization. In Vitro 20:404–408; 1984.
Author information
Authors and Affiliations
Additional information
The work was supported by grant AI-15748 from the National Institutes of Health, Bethesda, MD
Rights and permissions
About this article
Cite this article
McGarrity, G.J., Kotani, H. & Carson, D. Comparative studies to determine the efficiency of 6 methylpurine deoxyriboside to detect cell culture mycoplasmas. In Vitro Cell Dev Biol 22, 301–304 (1986). https://doi.org/10.1007/BF02623401
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF02623401