Detection of Mycoplasma salivarium Contamination in Cell Culture using PCR Method

Authors
V. Rostamizadeh 1
S. A. Pourbakhsh 2
E. Asli 2
A. Hadadi 1
1 Department of Microbiology, Karaj Branch of Islamic Azad University, Karaj, Iran
2 Mycoplasma Referance Laboratory, Razi Vaccine and Serum Research Institute, Karaj, Iran
Abstract
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 16S 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 62 cell culture samples were sent to the Mycoplasma Reference Laboratory at Razi Institute, Karaj, Iran for detection of Mycoplasma contamination. A total of 42 (67.75%) out of 62 samples scored positive according to the Mycoplasma genus. From these 42 samples, 15 (35.72%) reacted positively with a clear band of 434 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.

Keywords

 
 
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Pathobiology Reserach
Volume 16, Issue 3
December 2013
Pages 109-118