Volume 12, Issue 2 (Paramedical Sciences and Military Health (Summer 2017) 2017)                   Paramedical Sciences and Military Health 2017, 12(2): 13-20 | Back to browse issues page

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soheily Z, Soleimani M, Majidzadeh K. A PCR Assay for Detection of Mycoplasma Contamination in Cell Culture by 16S rRNA Specific Primers. Paramedical Sciences and Military Health. 2017; 12 (2) :13-20
URL: http://jps.ajaums.ac.ir/article-1-103-en.html
1- Department of Microbiology, Islamic Azad University, Qom branch, Qom, Iran
2- Department of Microbiology, Faculty of Medicine, AJA University of Medical Sciences, Tehran, Iran , soleimanidor@yahoo.com
3- Tasnim Biotechnology Research Center (TBRC), Faculty of Medicine, AJA University of Medical Sciences, Tehran, Iran
Abstract:   (2955 Views)

Introduction: Mycoplasmas are one of the most serious contaminants of cellular cultures and their biological productions. Mycoplasma diagnosis is conducted on the basis of culture and molecular methods. These methods are different from each other in terms of accuracy, reliability, and sensitivity. This study aimed to trace the mycoplasma contaminations in culture samples using 16S rRNA specific primers based on the PCR method.
Methods and Materials: 16S rRNA gene sequence of cell culture contaminant mycoplasmas were extracted from gen bank and were sorted using gene sequence pattern. Specific primers were designed using obtained synonymies. To build positive control structure, 16S rRNA gene proliferated based on the PCR methods then cloned in a plasmid vector.
Results: Electrophorus of PCR product of 16S rRNA revealed a band of 219bp length on Agarose gel. This pattern was not observed in negative control samples. Moreover, the fragment proliferation with the same size in recombinant plasmid confirmed the authenticity of cloning.
Discussion and Conclusion: Due to its high speed and accuracy, the method has potential for using in cell culture laboratories.

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Type of Study: Research | Subject: full articles
Received: 2017/06/14 | Accepted: 2017/07/30 | Published: 2017/09/21

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