A PCR Assay for Detection of Mycoplasma Contamination in Cell Culture by 16S rRNA Specific Primers

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.
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 Due to its high speed and accuracy, the method has potential for using in cell culture laboratories.