Mycoplasma contamination in human leukemia cell lines. I. Comparison of various detection methods.

The sensitivity and reliability of seven assays for mycoplasma detection were tested on a panel of leukemia cell lines. The assays used were: microbiological cultivation on broth and agar, immunofluorescent visualization of mycoplasmal DNA using DAPI (both direct staining and after multiplication of the contaminants on an indicator cell line), a nucleic acid hybridization assay with a radioactive probe specific for mycoplasmal rRNA, and ELISA with mycoplasma-specific polyclonal antibodies, a biochemical method utilizing 6-MPDR, and a mycoplasma-specific monoclonal antibody in immunofluorescence staining. The broth-agar method, the two DAPI tests and the RNA hybridization assay produced the highest detection rates; a number of false-negative cases were recorded by the other tests. The detection rates, costs, requirement for specialized equipment and other characteristics were evaluated for each method. Since each technique also has disadvantages and certain limitations and since no method can be regarded as the 'gold standard', at least two procedures should be used in routine screening for mycoplasma in cell cultures.