شناسايي عفونت مايكوپلاسمايي در رده هاي سلولي با روش PCR-ELISA

Detection of Mycoplasma Infection in Cell Cultures by PCR-ELISA

Abstract Background and objective: Mycoplasmas, particularly species 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 Mycoplasma species in cell lines by PCR-ELISA. Material and methods: 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 biotin. Results: 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 infected 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. Conclusions: Microbial culture and staining are common 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