Rapid isolation and detection of microbial contaminants in manufacturing

摘要

Pharmaceutical, biotech, and food/beverage industries predominantly use traditional cultured-based assays to detect microbial contamination in their products. There are significant benefits to the availability of faster methods that will provide equivalent or better results. Here, we present an approach to couple membrane filtration-based concentration and capture of organisms from complex liquid samples with purification and detection of the organism's nucleic acid using Real-Time Transcription-Mediated Amplification (Real-Time TMA; Gen-Probe Incorporated). This method can potentially be applied to detect a wide variety of micro organisms, including Mycoplasma, bacteria, fungi and viruses. Using a Mycoplasma assay we were able to detect 5CFU/mL of M. fermenians from a complex mammalian cell culture sample.