EP 2.6.7 (12.2) Now Mandatory: NAT Mycoplasma Testing Becomes Regulatory Standard

As of April 1, 2026, the revised European Pharmacopoeia chapter EP 2.6.7 (12.2) is now in effect, marking a significant shift in mycoplasma testing. Nucleic acid amplification techniques (NAT) are formally recognized as equivalent to conventional culture and indicator cell-based methods, redefining how mycoplasma detection is implemented in GMP environments. This transition not only enables faster and more sensitive detection, but also requires QC laboratories to adopt validated, compliance-ready NAT workflows to meet evolving regulatory expectations and support accelerated biopharmaceutical manufacturing.

Regulatory Shift: NAT Becomes an Equivalent Standard Under EP 2.6.7

Chapter European Pharmacopoeia 2.6.7 has long served as the central framework for mycoplasma detection in the European Pharmacopoeia, traditionally centered on culture-based and indicator cell culture methods. With the maturation and widespread adoption of molecular technologies such as quantitative PCR and digital PCR, the chapter has undergone a comprehensive revision. The updated framework formally integrates nucleic acid testing (NAT) as a primary approach, recognizing it as equivalent to conventional methods.

This shift reflects both advancements in microbial control science and increasing regulatory convergence across major pharmacopeias in Europe, the United States, and Asia. Notably, the revised chapter positions NAT not merely as an optional alternative, but within a risk-based testing strategy. In cases not covered by specific monographs, either conventional culture-based systems or NAT approaches may be applied, provided the selected method ensures detection of both culturable and non-culturable mycoplasma.

Core Regulatory Requirements for NAT-Based Mycoplasma Testing

The revision of EP 2.6.7 (Version 12.2) goes beyond introducing a new method---it establishes a comprehensive regulatory framework for nucleic acid testing (NAT), with each requirement directly addressing assay reliability and compliance. ACROBiosystems' mycoplasma detection solutions not only fully comply with the updated standards but also exceed key performance criteria, ensuring robust, future-ready quality control.

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Key Performance Advantages: Beyond Compliance

(1) Exceeding Regulatory Performance: Sensitivity and Specificity Assured

• Detects over 250 Mollicutes species, including all 10 strains required by EP. Achieves 100% detection at 10 CFU/mL across 24 replicates, with zero false negatives.

• No cross-reactivity with closely related organisms or cell lines (e.g., Streptococcus pneumoniae, E. coli, CHO/HEK293), ensuring 100% specificity and eliminating misidentification risk.

(2) Robust Across Complex Matrices: Built for Challenging Samples

With increased regulatory focus on complex matrices such as ATMPs and viral vectors, the ACROBiosystems solution is extensively validated for reliability:

• Compatible with high cell densities (10⁶--10⁷ cells/mL), 10% DMSO cryopreservation media, and antibiotic-containing cultures (e.g., puromycin, zeocin, G418).

• Proven performance in high-titer AAV (2×10¹³ vp/mL) and 50 mg/mL HSA matrices, delivering 100% spike recovery with stable Ct values---fully supporting cell and gene therapies as well as protein biologics.

(3) End-to-End Efficiency: Results in Under 3 Hours, Automation-Ready

• From sample extraction to result interpretation, the workflow is completed in just 2 hours 45 minutes---significantly faster than the typical 4--5 hour turnaround, accelerating batch release.

• Compatible with the Automated Nucleic Acid Extraction System (Cat. No. OPE-32S) and 96 V-Bottom Deep-well Plate (Cat. No. OPA-R013), enabling high-throughput processing while minimizing manual error.

(4) Comprehensive Compliance Support: From Validation to Implementation

• Includes a full method validation package covering six key parameters, such as limit of detection, precision, and robustness.

• Optional access to Eurofins Level 1 validation documentation for in-depth review.

• Manufactured in ISO 13485-certified facilities, each batch is accompanied by a complete Certificate of Analysis (COA), ensuring full traceability.

(5) Proven Stability: Flexible Storage and Reliable Performance

• Stable for up to 18 months at -15 to -30°C, and maintains performance for 20 days at 2--8°C.

• Demonstrates high consistency after 15 freeze--thaw cycles, with Ct value CV <1.02%, ensuring reliable performance during repeated use.

End-to-End Coverage: A Full-Spectrum QC Solution from R&D to Manufacturing

ACROBiosystems' mycoplasma detection solutions are not only aligned with evolving regulatory requirements but also support quality control across the entire biopharmaceutical lifecycle, addressing diverse application scenarios:

• Raw material testing: Early contamination screening for FBS, culture media, plasmids, and other inputs

• In-process control: Monitoring intermediate samples during bioreactor expansion and cell banking

• Batch release: Compliance testing for final products, including monoclonal antibodies, cell therapies, and viral vectors

• Specialized applications: Tailored detection for complex matrices such as ATMPs and gene therapy products

Mycoplasma Rapid Testing Workflow Solution

ACROBiosystems provides a comprehensive, integrated workflow for mycoplasma detection, including an Automated Nucleic Acid Extraction System (Cat. No. OPE-32S), Mycoplasma DNA Sample Preparation Kit (Magnetic beads) (Cat. No. OPA-E101) and Mycoplasma Detection Kit (qPCR) (Cat. No. OPA-S102). Built on the principles of regulatory alignment, performance beyond compendial standards, and full-scenario coverage, this solution is designed to be an optimal choice under the latest requirements. Existing reagents can be used without revalidation or upgrades, helping QC teams avoid repetitive qualification and preventing production delays.

Validated Performance Across Key Applications

• Mycoplasma testing solution is fully compatible with both cell and gene therapy workflows, covering common cell therapy matrices and gene therapy sample types.

Cell therapy matrices

Gene therapy sample

• High Sensitivity: Validated against pharmacopeial mycoplasma strains required by the EP, and USP, achieving a detection sensitivity as low as 1 CFU/mL.

Tested 10 mycoplasma standards (10 CFU/mL) in 24 replicates each; all returned positive---meeting/exceeding EP 2.6.7 standards (10 CFU/mL).

• High Specificity: No cross-reactivity.

Tested against two cell lines and four unrelated bacteria (e.g., Streptococcus pneumoniae, Lactobacillus acidophilus, Staphylococcus epidermidis, Bacillus subtilis)---all negative, confirming no cross-reactivity.

• High-quality assurance with superior performance compared to other competing products.

Eurofins validation

The mycoplasma rapid detection kit has been fully validated by Eurofins, a third-party organization. Results demonstrate strong comparability with the culture-based method, further confirming its reliability. To support platform development and regulatory submissions, complete validation reports are available upon request.

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