The Comparison of Microbial Culture and PCR Methods in Detection Of Cell Line To Mycoplasma

• Main Authors: Mohammad Reza Arabestani, Hossain Fazeli, Mahmmud Jedi Tehrani, Fazel Shokri
• Format: Article
• Language: fas
• Published: Vesnu Publications 2011-03-01
• Series: مجله دانشکده پزشکی اصفهان
• Subjects: Mycoplasma; Cell lines; Microbiological culture; PCR
• Online Access: http://jims.mui.ac.ir/index.php/jims/article/view/841

Background: Mycoplasma is a major contaminant of cell lines and is cosiderd as a serious problem of economic and biological importance in basic research, diagnosis, and biotechnology products. Detection of mycoplasma infection in cell cultures started on microbiological culture; later, other methods like DAPI staining and serological tests such as Indirect Immunofluorescnse, ELISA, DNA probe, PCR, PCR-ELISA, and Real-Timr PCR developed for detection of mycoplasma. Methods: In this study, a sensitive, specific, and rapid method was used for detection of varity of mycoplasma species in cell lines. This method was based on a PCR reaction using genus specific primers for 11 mycoplasma species. Results: Mycoplasma contamination using this assay was examind for 183 different cell line deposited in national cell bank of Iran. PCR showed that 48.6% of cell lines were contaminated with mycoplasma while 27.3% of them were found to be infected. In comparison to microbiological culture, PCR method was shown to be 100% sensitive and 70.7% specific. Conclusion: Our results using species specific primers reveald that the most important contaminating mycoplasma species in cell lines were mycoplasma fermentans, mycoplasma arginini, mycoplasma hyorhinis, and mycoplasma orale. We were also able to identify a number of cell lines which were contaminated whit more than one species of mycoplasma.