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

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...

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Main Authors: Mohammad Reza Arabestani, Hossain Fazeli, Mahmmud Jedi Tehrani, Fazel Shokri
Format: Article
Language:fas
Published: Isfahan University of Medical Sciences 2011-03-01
Series:مجله دانشکده پزشکی اصفهان
Subjects:
Online Access:http://jims.mui.ac.ir/index.php/jims/article/view/841
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author Mohammad Reza Arabestani
Hossain Fazeli
Mahmmud Jedi Tehrani
Fazel Shokri
author_facet Mohammad Reza Arabestani
Hossain Fazeli
Mahmmud Jedi Tehrani
Fazel Shokri
author_sort Mohammad Reza Arabestani
collection DOAJ
description 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.
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spelling doaj.art-88647adf0859478b8af423b05077010a2023-09-02T14:42:56ZfasIsfahan University of Medical Sciencesمجله دانشکده پزشکی اصفهان1027-75951735-854X2011-03-012812116761683569The Comparison of Microbial Culture and PCR Methods in Detection Of Cell Line To MycoplasmaMohammad Reza Arabestani0Hossain Fazeli1Mahmmud Jedi Tehrani2Fazel Shokri3PhD Student, Department of Microbiology, School of Medicine, Isfahan University of Medical Sciences, Isfahan, IranAssistant Professor, Department of Microbiology, School of Medicine, Isfahan University of Medical Sciences, Isfahan, IranProfessor, Department of Immunology, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, IranProfessor, Department of Immunology, School of Medicine, Tehran University of Medical Science, Tehran, IranBackground: 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.http://jims.mui.ac.ir/index.php/jims/article/view/841MycoplasmaCell linesMicrobiological culturePCR
spellingShingle Mohammad Reza Arabestani
Hossain Fazeli
Mahmmud Jedi Tehrani
Fazel Shokri
The Comparison of Microbial Culture and PCR Methods in Detection Of Cell Line To Mycoplasma
مجله دانشکده پزشکی اصفهان
Mycoplasma
Cell lines
Microbiological culture
PCR
title The Comparison of Microbial Culture and PCR Methods in Detection Of Cell Line To Mycoplasma
title_full The Comparison of Microbial Culture and PCR Methods in Detection Of Cell Line To Mycoplasma
title_fullStr The Comparison of Microbial Culture and PCR Methods in Detection Of Cell Line To Mycoplasma
title_full_unstemmed The Comparison of Microbial Culture and PCR Methods in Detection Of Cell Line To Mycoplasma
title_short The Comparison of Microbial Culture and PCR Methods in Detection Of Cell Line To Mycoplasma
title_sort comparison of microbial culture and pcr methods in detection of cell line to mycoplasma
topic Mycoplasma
Cell lines
Microbiological culture
PCR
url http://jims.mui.ac.ir/index.php/jims/article/view/841
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