Problems: Contamination of cell lines and biological products is one of the major problems of cell culture techniques. Rapid detection of mycoplasma contamination in cell culture is an important part of controlled laboratory. In order to existence of infected cells, leading to unreliable results, the need for a reliable method for laboratories is unavoidable. Polymerase chain reaction (PCR), a technique of rapid, sensitive and specific detection of bacteria is considered as a precise method to detect contamination. The aim of this study was to evaluate the efficacy of PCR in the detection of contaminants in cell cultures and other biological products.
Experimental approach: In this study, PCR techniques were optimized by use of MGSO and GPO-1 primers and16SrRNA target gene. Also the utilized PCR method was evaluated in terms of sensitivity and specificity. Finally, a simple DNA extraction and PCR of 164 cell culture of adipose tissue derived mesenchymal stem cells were done.
Findings: A 715 bp product was amplified by the primers and was confirmed by sequencing. The Reviews feature, with none of the tested DNA was amplified products. This method has a sensitivity limit of 10 copies of the target DNA. No cross-reactivity with genomic DNA of other microorganisms was observed.
Conclusion: The results of this study indicate that molecular methods based on polymerase chain reaction (PCR) have great importance due to their innate features. The observed results in this study are based on accuracy, speed, sensitivity and high specification of PCR technique according to conserved and common sequence of 16srRNA to detect Mycoplasma contamination in cell cultures.