Evaluation of three commercial kits for mycoplasma NAT assays: selection and quality improvement

Background Mycoplasma testing on cell lines or biological products used to be performed based on classical methods such as agar and broth medium and/or indicator cell culture. However, these methods require a long incubation period and are not adapted for samples, like live-attenuated vaccine viruses (which can not completely be neutralized) or cell therapy products (with short shelf life). NAT assays have several advantages including rapidtime to results, robustness and sensitivity. The European Pharmacopoeia updated the 2.6.7 section by adding the detection of Mycoplasma with NAT methods as an alternative to one or both classical methods. Texcell’s offers for Mycoplasma testing, which already included classical methods, were incremended with NAT assay. For this purpose, 3 commercial kits based on NAT assay were evaluated based of their claim to meet the European Pharmacopeia guidance for nucleic acid amplification techniques for Mycoplasma testing, including sensitivity and range of detection: CytoCheck from Greiner, MycoTOOL from Roche, MycoSEQ Mycoplasma detection kit from Life Technologies.