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Detecting Mycoplasma Cell Culture Contamination Using the CLARIOstar® Microplate Reader

Jacob Serfass and Megan Young, Penn State Hershey College of Medicine; Carl Peters, BMG LABTECH, Cary, NC, 05/2015

Mycoplasma are small-sized bacteria that are well-known as contaminants of laboratory cell cultures. Without having a cell wall around their membrane, they cannot be targeted with antibiotics. This makes them a big problem and often the whole culture is lost.

As mycoplasma can influence experimental results obtained for a certain cell line, cell culture labs need to check regularly if their cultures are contaminated by mycoplasma species. Here we show the application of the MycoAlert™ assay from Lonza to perform this test. The assay is specific for mycoplasmal enzymes that are not present in eukaryotic cells. After addition of substrate a high luminescent signal is created only in viable mycoplasma.

The luminescence detection was performed in a CLARIOstar® microplate reader. The instrument comes with the MARS Data Analysis software that allows the user to do calculations and validations to quickly identify samples which may be contaminated.