Rapid molecular assays for microbial contaminant monitoring in the bioprocess industry.

Adventitious contaminations with bacterial, viral, or fungal infectious agents represent a major risk associated with the manufacture and release of pharmaceutical products for human use, including vaccines, protein-based therapeutics, and antibodies. Early detection of contaminants in the biologicals production process might allow immediate action to correct such events without a significant interruption in the rate of production. Among the methods currently used for testing are cell culture, animal inoculation, electron microscopy, and in vitro molecular and antibody assays. Bacteria such as mycoplasma and mycobacterial species and most of the viral and fungal agents can take several days to weeks or even months to grow in culture. We have developed a broad-range microbial detection assay that uses the innovative Ibis biosensor platform, a rapid and high-throughput biosensor that is based on polymerase chain reaction (PCR) and electrospray ionization mass spectrometry to identify and quantify microbial contaminants. By combining the sensitivity of PCR with the accuracy of mass spectrometric detection, this technology generates a fingerprint that uniquely identifies an organism without a priori assumptions of the sample identity. This approach is capable of detecting known and unknown pathogens, as well as providing high-resolution genotyping and strain typing, and drug resistance and virulence information. Representative case studies are discussed here showing detection of minute virus of mice, mycoplasma, and an unknown virus that was identified as bluetongue virus.