Detection of Mycoplasmas in Cell Culture by Fluorescence Methods

The fluorescence stains used most often for mycoplasma detection are DNA binding fluorochromes (DNAFs) and fluoresceinated antibodies. Both permit direct visualization of individual mycoplasma organisms. DNAFs will bind to any appropriately conformed DNA that is present in a sample preparation and are, therefore, not specific for mycoplasmas. Conversely, fluoresceinated antibodies are highly specific for mycoplasmas, discriminating to species level. The two stains can be used individually or in combination, and they can be used either directly on the cells of interest or they may be used in a system involving mycoplasma-free indicator cells, which further enhances the diagnostic reliability of tests utilizing these fluorescent reagents.