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A chemiluminescent assay for the detection of mycoplasma contamination of cell cultures is described. Cells (and supernatant) derived from mycoplasma-contaminated cultures stimulate a burst of luminol-dependent chemiluminescence in cell suspensions containing phagocytic effector cell types. The assay conditions for spleen cells, human and bovine polymorphonuclear leucocytes as the responder or indicator cells have been optimized. The chemiluminescent assay can be utilized for both monolayer and suspension cell cultures and is more sensitive than colony formation on agar plates and electron microscopy. Results are obtained within 3-5 h including the time required for the preparation of the indicator cells. CL can be measured in the tritium window of standard liquid scintillation spectrometers after switching off the coincidence circuit.
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