A number of options are available for conclusive detection of the presence of mycoplasmas in contaminated cell cultures. A commonly but unreliable method is the DAPI staining of cells followed by microscopic analysis. However, PCR based methods give the most reliable results for routine checks. Biontex's (AFSBio's supplier) MycoSPY® and MycoSPY® Master Mix detection kits are rapid, reliable and highly sensitive methods of PCR-based mycoplasma detection in cell cultures.

The MycoSPY® kit contains a complete range of reagents for reliable and sensitive PCR. Each mycoplasma test kit contains Primer Mix, Internal Control, Taq Polymerase and a matched Taq polymerase buffer with nucleotide triphosphates (dNTPs). With some other products, the Taq polymerase must be purchased separately and is thus not matched to the buffer supplied in the kit, potentially impairing the sensitivity of detection results.

Literature:

1. J. J. Jung et al., Stem Cell Reports, 2014, 2: 592–605, doi.org/10.1016/j.stemcr.2014.03.006

2. Arash Yavari et al., Nature Communications, 2017, 8: 1258, DOI: 10.1038/s41467-017-01342-5

More than a quarter of all animal cell cultures are contaminated with mycoplasmas / mollicutes. The strains most commonly found in cell cultures, with a total probability of 94%, are: M. fermentans (47%), M. hyorhinis (19%), M. orale (10%), M. arginini (9%), A. laidlawii (6%) and M. hominis (3%). In addition, the following strains were found with lower probability: M. gallisepticum, M. pneumoniae, M. salivarium, M. synoviae and S. citri. All these Mollicute strains, in addition to a variety of other strains, are detected by MycoSPY®. BLAST analysis was used to verify of the primer specificity. The list shows only Mollicute strains with 100% primer match.