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 MycoSPY®, MycoSPY® Master Mix and MycoSPY® qPCR Master Mix detection kits are rapid, reliable and highly sensitive methods of PCR-based mycoplasma detection in cell cultures.

The MycoSPY® qPCR Master Mix consists of all necessary components for PCR analysis (hot-start Taq polymerase, buffer, dNTPs, primer, FAM/HEX fluorescence probes). The kit includes an Internal Control and PCR-grade water for rehydrating the lyophilized master mix. With a real-time PCR thermocycler suitable for detecting FAM and HEX fluorescent dyes, mycoplasma can be detected quickly and easily in cell cultures.

If mycoplasmas have been detected in your cell culture, the highly effective antibiotic mixture MycoRAZOR® eliminates the contamination rapidly and efficiently.

Workflow

The Master Mix with integrated FAM/HEX fluorescence probes is designed for maxiumum ease of pipetting and subsequent detection in real time:

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® qPCR Master Mix. BLAST analysis was used to verify of the primer specificity. The list shows only Mollicute strains with 100% probe and primer match.