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

Document Type : Original Article(s)

Authors

1 PhD Student, Department of Microbiology, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran

2 Assistant Professor, Department of Microbiology, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran

3 Professor, Department of Immunology, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran

4 Professor, Department of Immunology, School of Medicine, Tehran University of Medical Science, Tehran, Iran

Abstract

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.

Keywords

References

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Journal of Isfahan Medical School
Volume 28, Issue 121 - Serial Number 121
November and December 2011
Pages 1676-1683

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History

  • Receive Date: 20 March 2011
  • Revise Date: 01 November 2024
  • Accept Date: 06 April 2022

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