Detection of Mycoplasma Infection in Cell Cultures by PCR - ELISA

Mycoplasmas, particularly sp ecies of the genera Mycoplasma and Acholeplasma, are known to be occasional microbial contaminants of cell cultures that produce biologics. The aim of this study is to alternate a specific, sensitive and rapid method for detection of a variety of Mycoplasm a species in cell lines by PCR - ELISA. This method was based on a PCR - ELISA reaction using genus, specific primer and species specific primers for Mycoplasma species that labeled with Digoxigenine and specific probe, which labeled by b iotin. Mycoplasma contamination using PCR - ELISA was examined for 183 different cell line deposited in national cell bank of Iran. PCR - ELISA showed that 48.6 % of cell lines were contaminated whit Mycoplasma while 27.3% of them were found to be inf ected with microbial culture. PCR and PCR - ELISA methods, both of them showed the same results. In comparison to microbiological culture, PCR - ELISA method was shown to be 100% sensitive and 70.7% specific. Microbial culture and staining are com mon methods for detection of cell lines Mycoplasma contamination. These methods are time - consuming and so rapid contamination detection is critical for cell lines. For this reason PCR - ELISA method is recommended for rapid mycoplasma detection.