This test is a DNA fluorochrome staining assay coupled with co-culturing the test cell line in the presence of an indicator cell line such as Vero (a.k.a. indicator cell culture assay). After an incubation period of 3-5 days, the cells are fixed and stained with a DNA-binding fluorescent dye (Hoechst 33258 stain) and evaluated microscopically. This approach enhances the assay sensitivity by reducing background from genetically unstable cell lines (e.g. hybridomas, etc.) Other advantages of this assay include amplification of mycoplasma titers and detection of fastidious and “non-cultivable” organisms. Appropriate positive (< 100 CFU) and negative controls are processed with each sample.
Total Testing Time:
3-5 days
Sample Requirements:
Minimum Test Volume = 2 mL
This indirect method with the enrichment step results in a sensitivity of 100 CFU/mL, DNA staining methods are suitable for the mycoplasma screening of cell cultures and provide timely results.
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