Coriell Institute
Mycoplasma Testing

  1. Introduction
  2. Bibliography
  1. Introduction
    Because of the effects of mycoplasma contamination on growth and metabolism of cells in culture ( 1, 2, 3, 6, 11, 14) and the difficulty of detecting mycoplasma macroscopically, microbiologists and cell culturists have been eager to develop or adapt the most sensitive methods to assay for mycoplasma in cell cultures. The need for accurate and efficient detection of mycoplasma led to exploration of biochemical assays dependent on differences in metabolic pathways of mycoplasma and host cells (8, 10), molecular biological methods utilizing probes specific for mycoplasma (7, 13), the nonspecific staining of cytoplasmic DNA with Hoechst 33258 in cells contaminated with mycoplasma (5, 9), and PCR-based assays for mycoplasma recently (4, 12, 13).

    The Coriell Cell Repositories (CCR) has adopted a polymerase chain reaction (PCR) method developed by Applied Biosystems, the MycoSEQ® Mycoplasma Detection System (4460625, Life Technologies). The method is based on the similarities of mycoplasma 16S ribosomal DNA sequence and thus detects more than 90 known mycoplasma species in one multiplex, quantitative PCR. The assay is extremely sensitive, detecting as few at 10 copies of DNA per reaction and includes detection of PCR inhibition.

  2. Bibliography
    1. Barile, M.F.; Hopps, H.E.; Grabowski, M. 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: 35-46.
    2. Cole, B.C.; Naot, Y.; Stanbridge, E.J.; Wise, K.S. Interactions of mycoplasmas and their products with lymphoid cells in vitro. In: Razin, S.; Barile, M.F. eds. The Mycoplasmas, Vol. IV, Mycoplasma Pathogenecity. New York: Academic Press; 1985: 204-258.
    3. Fogh, J.; Holmgren, N.B.; Ludovici, P.P. A review of cell culture contaminations. In Vitro 7: 26-41; 1971.
    4. Hopert, A., Uphoff, C.C.; Wirth, M.; Hauser, H.; Drexler, H.G. Specificity and sensitivity of polymerase chain reaction (PCR) in comparison with other methods for the detection of mycoplasma contamination in cell lines. J. Immunol. Methods 164: 91-100; 1993.
    5. McGarrity, G.J. Detection of mycoplasmal infection of cell cultures. In: Maromorosch, K. ed. Advances in Cell Cultures. New York: Academic Press; 1982: 99-131.
    6. McGarrity, G.J.; Kotani, H. Cell culture mycoplasmas. In: Razin, S.; Barile, M.F. eds. The Mycoplasmas, Vol. IV. Mycoplasma Pathogenecity. New York: Academic Press; 1985: 353-390.
    7. McGarrity, G.J.; Kotani, H. Detection of cell culture mycoplasmas by a genetic probe. Exp. Cell Res. 163: 273-278; 1986.
    8. McGarrity, G.J.; Kotani, H.; Carson, D. Comparative studies to determine the efficiency of 6-methylpurine deoxyriboside to detect cell culture mycoplasmas. In Vitro 22: 301-304; 1986.
    9. McGarrity, G.J.; Steiner, T.; Vanaman, V. Detection of mycoplasmal infection of cell cultures by DNA fluorochrome staining. Methods in Mycoplasmology, II: 183-192; 1983.
    10. McGarrity, G.J.; Vanaman, V.; Sarama, J. Comparative studies between microbiological culture and uptake of uridine/uracil to detect mycoplasmal infection of cell cultures. Exp. Cell Res. 121: 159-165; 1979.
    11. McGarrity, G.J.; Vanaman, V.; Sarama, J. Cytogenetic effects of mycoplasmal infection of cell cultures: a review. In Vitro 20: 1-18; 1984.
    12. Rawadi, G.; Dussurget, O. Advances in PCR-based detection of mycoplasmas contaminating cell cultures. PCR Methods Appl. 4: 199-208; 1995.
    13. Razin, S. DNA probes and PCR in diagnosis of mycoplasma infections. Mol. Cell. Probes 8: 497-511; 1994.
    14. Robinson, L.B.; Wichelhausen, R.H.; Roizman, B. Contamination of human cell cultures by pleuropneumonia-like organisms. Science 124: 1147-1148; 1956.

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