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Mycoplasma are among the most common cell culture contaminants.

At a minimum, mycoplasma can alter normal cellular processes, virus production rates, and lymphocyte proliferation. Therefore, testing for mycoplasma in manufacturing cell substrates is essential. Moreover, mycoplasma can either directly or indirectly contribute to human disease, which represents a further regulatory concern. The general mycoplasma test is performed by using a method that tests for the presence of agar-cultivable and non agar-cultivable (cell-dependent / cell-associated) mycoplasma. Agar and semi-solid broth are used for detecting agar-cultivable mycoplasma species. An indicator cell line is used to detect non-agar-cultivable mycoplasma species. Mycoplasma are detected by their characteristic morphological pattern of fluorescence in the cell culture. Vero cells are typically used because they provide a large cytoplasmic area in which to view mycoplasmal growth. BioReliance offers a variety of mycoplasma assays designed to meet various testing and compliance and all are performed in accordance with US FDA, CFR and EP regulations. Points to Consider in the Characterization of Cell Lines Used to Produce Biologicals (1993 Points to Consider) and European Pharmacopoeia (EP) compliant assays are recommended. The EP requires that a test for inhibitory substances (i.e. mycoplasmastasis) be performed. BioReliance offers mycoplasmastasis studies to qualify each mycoplasma assay. The purpose of this assay is to detect the presence of stasis caused by the test article itself. The mycoplasmastasis study demonstrates that the test article is not inhibitory to the growth of mycoplasma that can be present in a test sample and can thereby limit the sensitivity of the assay. A mycoplasmastasis test does not have to be performed for each lot of material. This assay is typically performed on a representative lot of product or on a representative portion of a cell bank, provided that the manufacturing or cell banking process, sample preparation, and content of the sample do not change. Mycoplasmastasis testing is recommended for all licensed products. The presence of inhibitory substances can cause decreased assay sensitivity, leading to serious cell bank contamination, to manufacturing equipment or facility contamination, or to both. BioReliance also offers mycoplasma detection by PCR.