BioFire® assay blazes path to delivering mycoplasma results within three days or less

Jonathan Demick, PhD, Principal Scientist; Christopher Smith, Manager, Mycoplasma Services

Contamination of cell cultures and animal derived products by mycoplasma is a concern for biological and biopharmaceutical production. Mycoplasma contamination can have a myriad of impacts upon final products. To that end, regulatory guidance requires testing for the presence of mycoplasma in products that are derived from mammalian cell culture.

Traditional compendial testing for mycoplasma includes the direct culture and indicator cell culture methods. While both of these methods are the gold standard for mycoplasma detection, the timing for results is not conducive for newer autologous cellular therapies, as the direct method requires at least 28 days for completion.

The advent of nucleic acid technology (NAT) assays for the presence/ absence of mycoplasma reduced the time of analysis to within one week. Currently, the MycoSEQ™ assay meets all compendial limit of detection requirements. As with any PCR based method there is the risk of inhibition occurring. The Myco-SEQ assay targets DNA, which due to its half-life, does not produce results indicative of only viable organisms. While this assay is faster than the microbiologic assays, there are many steps from sample preparation, DNA extraction, PCR plating to analysis. As a way to minimize the large number of steps, a newer cartridge-based platform will be offered at Eurofins.

The BioFire^® assay encompasses the nucleic acid extraction, PCR plating and analysis within a single cartridge. This reduction in steps also provides a decrease in the duration of the test from start to finish. This system also targets RNA which has a shorter half-life than DNA. By targeting RNA, the probability of detecting viable organism will greatly increase, as RNA is only produced by viable organisms and may be present in higher concentrations due to amplification. This aligns a NAT method with compendial by having an increased likelihood of detecting viable organisms.

With the advent of autologous cellular therapies, testing time is valuable. With the BioFire system, mycoplasma results may be obtained within three calendar days. For some extremely tight timelines, 24-hour results can be achieved.