6th International Mycoplasma qPCR Testing User Day

Background & Objectives

The contamination of biopharmaceutical products, i.e. classic biologics, but also modern cell and gene therapy products with mycoplasmas, e.g. due to contamination of cell cultures during the manufacturing process, poses a potential health risk to patients. Mycoplasma can influence virtually every parameter of cell culture, often with only minor visible effects, leading to uncontrollable conditions that are undesirable in the pharmaceutical industry. Regulatory authorities therefore require manufacturers to test their biopharmaceutical products and ensure that the released products are free of mycoplasma. Most regulatory authorities have issued guidelines that include protocols for mycoplasma testing, and some provide recommendations for the validation of rapid NAT (nucleic acid amplification techniques) testing methods. This provides you with a scientifically sound introduction to the field of rapid mycoplasma testing with a special focus on NAT and, in particular, qPCR methods. It includes lectures, case studies and interactive roundtable discussions by users for users.

Target Audience

The Pre-Conference Workshop is directed to responsible personnel involved in Quality Control testing of biopharmaceuticals and biologics, e.g.:

• QC Director, Manager, Scientists, Microbiologists, and Process Microbiologists
• Analytical Experts
• Biosafety and Pathogen Safety departments
• Bioassay Developers
• Regulatory departments

It is also useful for service providers, such as contract research organisations and contract manufacturers.

Register now

Programme

Mycoplasma Testing – Authorities Point of View
Jan-Oliver Karo, Paul-Ehrlich Institut, German Federal Institute for Vaccines and Biomedicines

• Regulatory Background
• Expectations, Experiences and New Developments

Nucleic Acid Testing for Mycoplasma Detection: From Regulatory Requirements to GMP Implementation
Dr Marc Meichenin, CleanCells
Dr Caroline Kassim Houssenaly, bioMérieux

• A comparison with traditional compendial methods (EP 2.6.7, USP <63>)
• Regulatory expectations, including the new European Pharmacopoeia guidelines (EP 12.2, April 2026)
• The importance of matrix validation and genome copy–based sensitivity thresholds
• Case study examples from QC environments highlighting challenges and solutions during method validation and routine integration

Mycoplasma Platform for Rapid QC in Short-Shelf Life ATMPs
Dr Jonathan Hanley, Shimadzu

• Mycoplasma Platform for Rapid QC in short-shelf life ATMPs
• Mycoplasma detection compliant with EP 2.6.7, JP17, USP63,
• Rapid DNA extraction and PCR in line with the needs of short shelf life ATMPs
• Mycoplasma detection limit below 10 CFU/mL in a background of high cell density (108 human/mammalian cells)

Leveraging the Capabilities of qPCR in Rapid Mycoplasma Validation Study Design and Execution including a Case Study on Application in Investigation of a Contamination Event
Michael Brewer, Thermo Fisher Scientific

• Fundamentals and application of the quantitative performance of qPCR in method development
• Definition of objective acceptance criteria and qualification of Mycoplasma spiking stocks
• Application of qPCR in the assessment of viability and contamination risk
• Design of a validation study and results of a 3-arm study to evaluate the detection limit of 2 qPCR methods and comparability with the USP <63> Mycoplasma test
• A case study on the use of qPCR in investigating a contamination incident in a CHO bioreactor, which includes the application of quantitative values to determine the root cause

Guidelines for Choosing the Appropriate Mycoplasma Standards for Method Validation by NAT
Yoann Mainguy, Merck

• Current Guidelines
• Why is the concept of the GC/CFU ratio crucial in the validation of a method for the detection of mycoplasmas?
• How to calculate this ratio, and what are the associated challenges?
• What are the optimal characteristics that mycoplasma standards should provide for validation purposes?

Optimization and Implementation of the MycoSEQ SYBR and TaqMan Kits for Lot Release
Joleen Simpson, Lilly

• Rapid PCR-based mycoplasma detection methods can replace the traditional 28-day culture-based testing.
• Thermo Fisher's MycoSEQ kits (both SYBR Green and TaqMan) enable the rapid detection of over 90 species of Mycoplasma, but we encountered significant PCR inhibition with their original SYBR Green kit.
• We were able to optimize the sample preparation to work with our CHO products, but challenges with the MycoSEQ discriminatory positive control led us to transition to the TaqMan platform.
• The MycoSEQ Plus TaqMan kit is more robust for our applications and requires less modifications. The development and implementation steps for kits will be discussed.

Bridging Speed and Sensitivity: Validation of a Hybrid Mycoplasma Detection Method for Complex Product Matrices
Dr Lori Daane, Bionique Testing Laboratories

• Benefits of hybrid PCR-based methods compared to traditional PCR methods, especially for complex product matrices.
• Important considerations for evaluating hybrid and other alternative methods including:
  • selecting an alternative method,
  • identifying test article volume and turnaround time constraints,
  • utilizing platform validation data, and
  • demonstrating parameters for method suitability
• Case study data on complex product matrices will be presented

Implementing a New Positive Control
Karen de Roy, J&J

Summary, Q&A and Plenum Discussion
Alexander Barthes, Roche