microbial-control-monitoring-strategies-for-cleanroom-environments

Microbial Control and Monitoring Strategies for Cleanroom Environments and Cellular Therapies

A well-validated and holistic program that incorporates robust gowning, cleaning, environmental monitoring, and personnel monitoring measures is critical ...

National Institutes of Health

Testing of cellular therapy products for  is a regulatory requirement by the United States Food and Drug Administration (FDA) to ensure the sterility and safety of the product prior to release for patient infusion. The risk of  contamination in cell culture is high. Gold standard testing follows USP <63> which requires a 28-day agar and broth cultivation method that is impractical for short shelf-life biologics. Several commercial molecular platforms have been marketed for faster raw material and product release testing; however, little performance data are available in the literature. In this study, we performed a proof-of-principle analysis to evaluate the performance of five commercial molecular assays, including the MycoSEQ  detection kit (Life Technologies), the MycoTOOL  real-time detection kit (Roche), the VenorGEM qOneStep kit (Minerva Biolabs), the ATCC universal  detection kit, and the Biofire  assay (bioMérieux Industry) using 10 cultured  spp., with each at four log-fold dilutions (1,000 CFU/mL to 1 CFU/mL) in biological duplicates with three replicates per condition ( = 6) to assess limit of detection (LOD) and repeatability. Additional testing was performed in the presence of tumor infiltrating lymphocytes (TILs). Based on LOD alone, the Biofire  assay was most sensitive followed by the MycoSEQ and MycoTOOL which were comparable. We showed that not all assays were capable of meeting the ≤10 CFU/mL LOD to replace culture-based methods according to European and Japanese pharmacopeia standards. No assay interference was observed when testing in the presence of TILs.

Amanda D. East

Sterility Testing Service,
Department of Laboratory Medicine,
Clinical Center,
Sterility Testing Service, Department of Laboratory Medicine, Clinical Center

National Institutes of Health

Microbial Control and Monitoring Strategies for Cleanroom Environments and Cellular Therapies

Amanda D. East

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National Institutes of Health

Microbial Control and Monitoring Strategies for Cleanroom Environments and Cellular Therapies

Sterility Testing Service,
Department of Laboratory Medicine,
Clinical Center,
Sterility Testing Service, Department of Laboratory Medicine, Clinical Center