Mycoplasma testing

Mycoplasma testing

Mycoplasma species are often found as common contaminants in cell cultures, virus stocks, and other cell-derived biologicals. Because of their small size (~ 0.15 µm) and resistance to common antibiotics such as penicillin and streptomycin, contamination of cell cultures usually occurs unnoticed. However, mycoplasma can cause adverse effects on cells in culture including changes in growth, morphology, metabolism, protein synthesis, and virus replication.

BaseClear uses a validated PCR test for rapid and reliable detection of Mycoplasma contamination in various in situ biologicals including cell cultures and virus stocks. Between 5 and 20 samples per order are tested by us together with internal, positive, and negative controls.

Testing for mycoplasma infection is now an essential requirement in reaching quality control standards for laboratories concerned with research, production and maintenance of cell lines. Since mycoplasma contamination is a recurring problem that can cause non-reproducible results in research and productivity losses in industry, BaseClear recommends routine diagnostic testing of cell cultures on a monthly basis as well as before cell-banking and cryopreservation.

The test is based on specific amplification of conserved regions in the 16S rRNA genes of the mycoplasma genome. This allows for detection of all mycoplasma, acholeplasma, and ureaplasma species tested so far and usually encountered as contaminants in cell cultures.

Using this highly sensitive test, we can detect as little as 2-5 mycoplasma particles of more than 25 different mycoplasma species, including M. fermentans, M. pirum, M. orale, M. hominis, M. argininii, M. pneumoniae, Acholeplasma laidlawii and other mycoplasma species usually encountered as contaminants in cell cultures.

Cell lines should be cultured in the absence of antibiotics for 3 days without refreshment of culture media. If cells are cultured for a longer period, substances may accumulate in the medium that inhibit the test.

Per sample, collect 1 ml of culture supernatant in 1.5 ml Eppendorf Safe-Lock tubes or screw cap tubes. To maximize test sensitivity, supernatants should be collected from 90-100% confluent cell cultures.

The minimum amount of samples per order is 5.