Detection of Mycoplasma salivarium Contamination in Cell Culture using PCR Method

Objective: Mycoplasma salivarium (M. salivarium) isone of the most common contaminants present in cell culture laboratories that cause undesirable effects on cell cultures. Thus, the identification and rapid diagnosis in controlling and prevention of this contaminant are important. The aim of this study is the detection of Mycoplasma salivarium contamination in cell culture using polymerase chain reaction (PCR) method. Methods: A ۱۶S rRNA-based Mycoplasma genus and specific primer PCR method for M. salivarium was developed. The sensitivity and specificity of this method were determined. The PCR test was used after we extracted DNA from the cultured isolates. Results: A total of ۶۲ cell culture samples were sent to the Mycoplasma Reference Laboratory at Razi Institute, Karaj, Iran for detection of Mycoplasma contamination. A total of ۴۲ (۶۷.۷۵%) out of ۶۲ samples scored positive according to the Mycoplasma genus. From these ۴۲ samples, ۱۵ (۳۵.۷۲%) reacted positively with a clear band of ۴۳۴ bp in the M. Salivarium-specific PCR method. Conclusion: Due to the high percentage of M. salivarium contamination in cell cultures, we recommend aseptic conditions be used in the laboratory when working with cell cultures. The PCR method is a suitable and valuable tool for the detection of M. salivarium contamination in cell cultures with appropriate and specific primers. This PCR method can be processed in less than one day.

کلیدواژه ها:

PCR ، PCR ، Contamination ، Mycoplasma salivarium ، cell culture ، مایکوپلاسما سالیواریوم ، آلودگی ، کشت سلول