Combining a Respiration-Based Method with Universal qPCR for 3-Day Rapid Sterility Testing

Area of Science:

• Microbiology
• Molecular Biology
• Biotechnology

Background:

• Sterility testing is critical for pharmaceutical products, especially cell-based therapies and DNA preparations.
• Traditional sterility methods can be time-consuming, delaying product release.
• Rapid methods are needed to accelerate release and ensure patient safety.

Purpose of the Study:

• To develop and validate a rapid sterility testing method for cell-therapy drug products and E. coli DNA plasmid preparations.
• To reduce the time to detection for sterility testing.
• To establish a sensitive and reliable method for sterile release assays.

Main Methods:

• Combined automated BacT/ALERT (bioMérieux) enrichment with PCR detection using the ATMP sterile release qPCR assay (Sartorius).
• Developed sample preparation protocols for non-filterable cell-therapy products and cell-free DNA plasmids.
• Included steps for removal of contaminating DNA, PCR inhibitors, and physical lysis.

Main Results:

• Achieved a time to detection of 3 days for sterility testing.
• Demonstrated a limit of detection (LOD) of 1 CFU (Colony Forming Unit).
• Presented validation data for the BacT/ALERT system and pre-validation experiments for the full assay.

Conclusions:

• The developed rapid sterility method is effective for both cell-therapy products and DNA plasmid preparations.
• This method offers a significant reduction in testing time compared to traditional approaches.
• The assay supports efficient sterile release of critical biopharmaceutical products.