Related Experiment Video
Updated: Jun 16, 2026
A Whole Cell Bioreporter Approach to Assess Transport and Bioavailability of Organic Contaminants in Water Unsaturated Systems
Published on: December 24, 2014
Comprehensive Study Identifies a Sensitive, Low-Risk, Closed-System Model for Detection of Fungal Contaminants in
1Microbiology Service, National Institutes of Healthgrid.94365.3d, Bethesda, Maryland, USA.
Abstract:
The U.S. Food and Drug Administration (FDA) regulates manufacturing and testing of advanced therapeutic medicinal products (ATMPs) to ensure the safety of each product for human use. Gold-standard sterility testing (USP<71>) and alternative blood culture systems have major limitations for the detection of fungal contaminants. In this study, we evaluated the performance of iLYM (lactic acid-fermenting organisms, yeasts, and mold) medium (designed originally for the food and beverage industry) to assess its potential for use in the biopharmaceutical field for ATMP sterility testing. We conducted a parallel evaluation of four different test systems (USP<71>, BacT/Alert, Bactec, and Sabouraud dextrose agar [SDA] culture), three different bottle media formulations (iLYM, iFA Plus, and Myco/F Lytic), and two incubation temperatures (22.5°C and 32.5 to 35°C) using a diverse set of fungi (n = 51) isolated from NIH cleanroom environments and previous product contaminants. Additionally, we evaluated the effect of agitation versus delayed-entry static preincubation on test sensitivity and time to detection (TTD). Overall, delayed entry of bottles onto the BacT/Alert or Bactec instruments (with agitation) did not improve test performance. USP<71> and SDA culture continued to significantly outperform each automated culture condition alone. However, we show, for the first time, that a closed-system, dual-bottle combination of iLYM 22.5°C and iFA Plus 32.5°C can provide high fungal sensitivity, statistically comparable to USP<71>, when tested against a diverse range of environmental fungi. Our study fills a much-needed gap in biopharmaceutical testing and offers a favorable testing algorithm that maximizes bacterial and fungal test sensitivity while minimizing risk of product contamination associated with laboratory handling.
Insights
This study introduces a novel dual-bottle method using iLYM and iFA Plus media for advanced therapeutic medicinal product sterility testing. This approach offers high fungal sensitivity, comparable to gold standards, improving biopharmaceutical safety.
Area of Science:
- Biopharmaceutical Manufacturing
- Microbiology
- Quality Control
Background:
- Current sterility testing methods for advanced therapeutic medicinal products (ATMPs) face limitations in detecting fungal contaminants.
- Gold-standard USP<71> and alternative blood culture systems have significant drawbacks for comprehensive fungal detection.
Purpose of the Study:
- To evaluate the performance of iLYM medium, originally for food industry use, for ATMP sterility testing.
- To assess a novel dual-bottle system for enhanced fungal detection in biopharmaceutical products.
Main Methods:
- Parallel evaluation of USP<71>, automated systems (BacT/Alert, Bactec), and SDA culture.
- Testing of iLYM, iFA Plus, and Myco/F Lytic media formulations at different temperatures (22.5°C and 32.5-35°C).
- Assessment of fungal isolates (n=51) from cleanroom environments and product contaminants, including agitation vs. delayed-entry incubation effects.
Main Results:
- Automated systems alone did not outperform USP<71> or SDA culture.
- Delayed entry incubation did not improve automated system performance.
- A closed-system, dual-bottle combination of iLYM at 22.5°C and iFA Plus at 32.5°C demonstrated high fungal sensitivity, statistically comparable to USP<71>.
Conclusions:
- The iLYM/iFA Plus dual-bottle system offers a promising alternative for ATMP sterility testing, addressing limitations of current methods.
- This novel algorithm enhances fungal detection sensitivity while minimizing contamination risks associated with laboratory handling.
- The findings provide a valuable solution for biopharmaceutical quality control and product safety assurance.
