New Validation Guide for Mycoplasma Testing according to EP 2.6.7 and USP <77>

Do you need to perform a method suitability test even if your mycoplasma PCR assay has already been validated by the manufacturer? Which standards and controls should be included? And how can you estimate the material required for your validation study?

The updated European Pharmacopoeia chapter EP 2.6.7 and the new USP <77> introduce important requirements for nucleic acid amplification techniques (NAT) used in mycoplasma testing. Even when a PCR assay has been extensively validated by the manufacturer, users must still demonstrate that the method is suitable for their specific product matrix. EP 2.6.7 refers to this assessment as the “Test for Inhibitory Substances”, while USP <77> describes it as the “Method Suitability Test”.

Our new Validation Guide for Mycoplasma Testing provides a practical overview of how this product-specific method suitability assessment can be planned and performed.

What does the Validation Guide include?

The guide summarizes the key regulatory expectations and explains which tests are required before a NAT-based mycoplasma assay can be introduced into routine testing.

It covers:

• the regulatory framework of EP 2.6.7 and USP <77>
• suitable mycoplasma standards for validation and routine testing
• the selection of relevant mycoplasma species
• a practical proposal for product-specific validation
• the recommended experimental setup and acceptance criteria
• the controls to include in the workflow
• example calculations for the required extraction and PCR reactions and standards

The guide includes example validation setups for both four and ten mycoplasma species and illustrates how samples, negative extraction controls, external positive controls, no-template controls and PCR-positive controls can be integrated into the workflow.

Who is the guide intended for?

The Validation Guide is designed for laboratories that use NAT-based methods for mycoplasma testing in cell cultures, biopharmaceutical products, vaccines or advanced therapy medicinal products.

It can help answer practical questions such as:

• Which mycoplasma reference material is suitable for my validation?
• How should the method suitability study be structured?
• Which controls should be included?
• How much material will be required?

The aim of the guide is to make the regulatory expectations easier to translate into a practical laboratory workflow.

It can support laboratories in estimating the required materials, defining suitable controls and preparing a structured validation plan for their individual sample matrix.

The document focuses on the use of the Venor® Mycoplasma qPCR, Venor® Mycoplasma dPCR, and Venor® Mycoplasma Extraction kit together with the new 100GC®, 10CFU®, and 100CFU® Mycoplasma Standards.

Download the Validation Guide

Download the complete Validation Guide for Mycoplasma Testing according to EP 2.6.7 and USP <77> to review the proposed validation strategy, experimental setup and material requirements.

Please note: The guide is intended for informational purposes and provides a general recommendation. It does not replace the official pharmacopoeial chapters or binding regulatory advice. The final validation strategy should be assessed for the individual product and discussed with the relevant regulatory authority.