Journal des sciences vétérinaires et du diagnostic médical

Detection of Mycoplasma Spp. in Cell Cultures by Genus-Specific Polymerase Chain Reaction Protocol

Claudia F. Lobos, María A. Martínez and Carlos O. Navarro

Contamination of cell cultures with Mycoplasma spp. complicates the basic investigation and the development of biological products. The effects of these bacteria on cultivated cells are changes in the metabolism, immunological and biochemical properties, growth, viability, etc. The Mycoplasma spp. infection on cell cultures might
not be detected by visual inspection or common microscopy. Hence, it is important to go through routine periodic evaluations with a highly sensible and highly specific fast method. Regarding the previous statement, this memoir was based on the molecular diagnosis of Mycoplasma spp., by detecting the 16S rRNA gene through the conventional polymerase chain reaction technic, on cell culture samples from different laboratories of the University of Chile and the Institute of Public Health of Chile. The results obtained in positive controls as in negative controls, allowed the validation of this method in the Faculty of Veterinary Sciences and by applying it on suspicious samples from the Institute of Biomedical Sciences of the University of Chile. This finding was verified by alignment of nucleotide sequences using the Clustal Ω and BLAST software, both online freeware, giving a 97% of nucleotide identity percentage respect to Mycoplasma spp. from the GeneBank®.