Survey of mycoplasma infections in cell cultures and a comparison of detection methods

A total of 1424 cell cultures was assayed for mycoplasmas by microbiological culture and fluorescent DNA staining. Of these cultures, 412 (29%) were infected with mycoplasmas. The most frequently occurring mycoplasma species were Mycoplasma orale (34%), M. hyorhinis (26%), M. arginini (21%), M. fermentans (13%) and Acholeplasma laidlawii (5%). A few isolates each of M. hominis, M. pulmonis and M. bovis were also detected. When detection methods were compared, microbiological culture produced false-negative results for 0.7% (3 of 412) of the infected cell cultures. DNA staining performed directly on the cells was falsely negative in 2.4% (5/207) of the mycoplasma-infected cultures that were compared, DNA staining performed on indicator cells was falsely negative in 3.1% (7/226). False positives appeared in direct DNA-staining in 1.8% (7/386) of the mycoplasma-free cultures and with DNA staining on indicator cells in 0.5% (3/620). For 11% of the cell cultures, the reading of the DNA staining was ambiguous. With DNA staining on indicator cells, 10% of the test results were ambiguous, but by further passage and staining on new indicator cells it was possible to get a definite diagnosis.