Journal of Veterinary Science & Medical Diagnosis ISSN: 2325-9590

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Research Article, J Vet Sci Med Diagn Vol: 7 Issue: 2

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

Claudia F. Lobos1, María A. Martínez2 and Carlos O. Navarro1*

1Animal Preventive Medicine Department, Microbiology Laboratory, Faculty of veterinary sciences and livestock, University of Chile, Chile

2Microbiology Program ICBM, Medicine Faculty, University of Chile, Chile

*Corresponding Author : Carlos O Navarro
Professor of Biochemistry, Faculty of veterinary sciences and livestock, University of Chile, Chile
Tel: + (56 2) 2978 5627
E-mail:
[email protected]; [email protected]

Received: May 05, 2018 Accepted: May 16, 2018 Published: May 21, 2018

Citation: Lobos CF, Martínez MA, Navarro CO (2018) Detection of Mycoplasma Spp. in Cell Cultures by Genus-Specific Polymerase Chain Reaction Protocol. J Vet Sci Med Diagn 7:2. doi: 10.4172/2325-9590.1000256

Abstract

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®. 

Keywords: Cell cultures; Mycoplasma spp.; 16S rRNA gene; PCR

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