1. Department of biology –Faculty of science, East Tehran Branch -Islamic Azad University –Tehran / Iran 2. Department of Microbiology – shahr-e-Qods Branch – Islamic Azad University –Iranian Gene Fanavar Institute (IGF), Tehran 3. Department of biology –Faculty of science, East Tehran Branch -Islamic Azad University –Tehran / Iran
Mycoplasmas are the major contributors to biological products and cell lines throughout the world and are easily able to pass through bacteriological filters. mycoplasma hyorhinis is one of the most important factors in cell culture infections which is the source of contamination of commercial animal serums used in cell culture. therefore, identification and diagnosis of these microorganisms is important in order to prevent significant damage resulting from the contamination of these microorganisms and need for a reliable method to diagnose this species from mycoplasma in manufacturing centers.
the purpose of this study was to set up and apply polymerase chain reaction method as a rapid, sensitive, and specific method for detecting mycoplasma hyorhinis in cell culture and biological contamination.
In this study, 100 suspected mycoplasma cultivars isolated from biological products on pplo-broth were collected from the razi institute and then dna samples were extracted by dng plus method. after of optimization, specificity and evaluation of limit of detection, test carry out on all of the samples.
The product of 604 bp amplicon was observed at 1.5% agarose gel. in the specificity test all of microorganism’s dna were negative and just observed with mycoplasma hyorhinis dna. from 100 samples tested, 63 samples (63%) were positive.
due to the high rate of cell culture contamination with mycoplasma hyorhinis and the specificity, accuracy and sensitivity of the pcr test, as well as the time and cost of other methods, using of this method is recommended for detection of biological products in production facility.