Development of DNA extraction and PCR amplification protocols for detection of Mycoplasma bovis directly from milk samples.

Mycoplasmas are micro-organisms that are widely distributed in nature and they are able to colonise animals, humans, plants and soil. Mycoplasma spp are grouped under the class Mollicutes (Bsoft skin^) and they differ phenotypically from other bacteria for their small dimensions (0.2j1 mm) and by the fact that they do not have the genetic ability to produce a cell wall. Between pathogenic species for animals, Mycoplasma bovis has a determinant role in many bovine diseases. This organism is a causative agent of mastitis, arthritis, pneumonia, otitis media, abortion, subcutaneous abscesses and fertility disorders. The diagnosis of Mycoplasma bovis is most commonly made by microbiological procedures, using different specific growth media and long incubation times. Thus, isolation and identification of Mycoplasma bovis are notoriously difficult and time consuming. The use of DNA-based methods for Mycoplasma bovis identification, based on amplification of specific, highly-conserved genes, reduces the assay time to hours as opposed to days or weeks, with high sensitivity and specificity.