1. 1Department of Microbiology – shahr-e-Qods Branch – Islamic Azad University –Tehran / Iran 2. Department of Microbiology – shahr-e-Qods Branch – Islamic Azad University –Tehran / Iran 2Iranian Gene Fanavar Institu 3. Department of Microbiology – shahr-e-Qods Branch – Islamic Azad University –Tehran / Iran
mycoplasmas are one of the most important pollutants in biological products and cell cultures, and are also one of the most dangerous destructive factors for biological outcomes. one of the most common contaminating factors in cell cultures and biological products is mycoplasma arginini, the source of which is usually in serum of infected fetal bovine or the serum of infected newborn bovine. therefore, identification of this agent and speed of detection in control and prevention contamination of biological products is important.
to use a rapid and specific method for detection of mycoplasma arginini contamination in biological products and cell cultures by polymerase chain reaction (pcr).
in this study, 100 samples of positive culture were collected from the razi institute in a pplo-broth medium, isolated from biological products, cell culture media and other components. pcr test diagnosis of mycoplasma arginini was optimized by using standard strain and was tested for specificity and sensitivity. dna samples were extracted by dng-plus method, and the pcr test was performed with positive and negative control.
amplicon 545 bp was amplified by using mycoplasma arginini of specific primers and was detected in 1.5% agarose gel electrophoresis.
in specificity test only with dna of mycoplasma arginini amplicon was showed . from 100 tested samplaes, 3 positive samples with positive and negative controls were observed.
this study showed that a number of positive pplo-broth cultures are contaminated by mycoplasma arginini. since other methods seen to be time-consuming and costly, the use of pcr testing, which has high accuracy and sensitivity, is recommended for the detection of contamination of biological products and cell cultures.