Rapid molecular detection of the biological products contamination by Mycoplasma orale

Introduction: Mycoplasmas are one of the most serious pollutants of the biological products and cell cultures and it appears that they are one of the factors destroying the biological results. Among them, Mycoplasma orale is one of the five basic Mycoplasmas polluting the cell cultures, biological products and middle and final products which are present in upper part of the human respiratory system.Purpose: Optimization of the rapid detection test for Mycoplasma orale and evaluation of the contamination percent in biological products.Methodology: 100 PPLO-Broth culture samples suspect for Mycoplasma contamination were collected from Razi institute and then, their DNA was extracted using DNG PLUS method. PCR test was optimized with using 16SrRNA as a target gene and evaluated from the respect specificity and limit of detection. Then, optimized test was conducted on all samples along with positive and negative controls.Results: PCR product with the size of 583 bp observed in 1.5% agarose gel. In investigation of the specificity of primers, only formed the intended bands with Mycoplasma orale DNA. From 100 tested samples, only nine (9%) were positive.Conclusion: according to studies, it was found that PCR method is an appropriate method for detection of Mycoplasma orale. While a high percent of contamination of the biological products is due to of this agent.

Keywords:

Mycoplasma orale , biological products , contamination , Molecular detection , PCR