Compliant Mycoplasma Testing: New Regulatory Standard for NAT Methods
The revised European Pharmacopoeia chapter 2.6.7 "Mycoplasmas" (version 12.2) introduces an updated regulatory framework for nucleic acid amplification techniques (NAT), formally recognizing them as equivalent alternatives to culture-based methods. The revision aligns European requirements with the Japanese and United States pharmacopoeias, supporting global harmonization in mycoplasma testing. For GMP-regulated laboratories, this update represents an important step toward consistent, internationally accepted testing strategies for biopharmaceutical and advanced therapy manufacturing.
Key Technical Requirements
The revision establishes clear sensitivity criteria when NAT methods replace culture-based assays. The required limit of detection is ≤ 10 CFU/mL or < 100 genomic copies per milliliter. Using genomic copies as a standardized unit improves comparability between molecular and culture-based results.
Testing must detect both culturable and non-culturable mycoplasmas and cover both cell-associated and cell-free fractions, reflecting the biological behavior of these organisms.
The revised chapter requires validation of the test method in the user's specific product matrix, even when commercially validated assays are employed. This ensures adequate sensitivity and confirms the absence of inhibitory effects under actual testing conditions.
Updated specifications for reference materials define a genomic copy to CFU ratio below 10, improving traceability. NAT systems must also include defined internal, positive, and negative controls, with the positive control set close to the assay's detection limit.
Implementation Timeline
The revised chapter has been published and becomes mandatory in April 2026. Quality control laboratories should align their testing strategies with the new requirements in a timely manner. NAT-based methods, successfully applied for many years, are now fully integrated into the regulatory framework and offer a reliable, compliant approach for mycoplasma detection.
