710345Mycoplasma screening - bringing invisible parasites to lightClosedGRD Proof of ConceptInnovate UKMycoplasma belong to the class Mollicutes which are the smallest free living organisms known. They are derived from Gram negative bacteria. They are commonly found in the oral cavity and as symbiotic gut flora. Some mycoplasma will invade host tissues and cells and replicate intracellularly where they can affect the growth and morphology of the cell and can alter its biochemical, metabolic and immunological functions. Mycoplasma have been implicated in a variety of diseases including Chronic Fatigue Syndrome, pneumonia and arthritis. The contamination of mammalian cell cultures by mycoplasma has become widely recognised as a major problem in cell based research and drug discovery. It is estimated that up to 35% of all mammalian cell cultures are infected by mycoplasma. Many journals and grant giving bodies are now stipulating that all cell based research is shown to be mycoplasma free by routine testing before research will be accepted for publication or as part of the terms of a research grant. To date, routine screening methods for mycoplasma contamination have either been lengthy (culture taking 14 to 28 days), required experienced personnel, been difficult to interpret or been unreliable (giving false negative or false positive results). As the basis of this project an assay has been designed based on the detection of a specific mycoplasma enzyme the activity of which is coupled to a bioluminescent detection system allowing for the rapid and sensitive detection of mycoplasma contamination. The assay is designed to be simple to perform and with easily interpreted results requiring only a single reagent addition with results obtained in less than 60 minutes. With this assay any sample tested that contains mycoplasma will emit light where the amount of light emitted is proportionate to the number of live mycoplasma present.