RAPID DETECTION OF MYCOPLASMA-BOVIS IN MILK SAMPLES AND NASAL SWABS USING THE POLYMERASE CHAIN-REACTION

Two methods are suggested that allow direct detection of Mycoplasma bo vis from clinical samples, i.e. milk and nasal swabs, respectively. Mi lk samples were trypsinized in the presence of Triton X-100 and passed through a DNA-binding filter membrane, from which DNA was extracted a nd subjected to PCR. The detection limit was 500 cfu ml(-1) on agarose gels and 50 cfu ml(-1) after Southern hybridization so that the metho d can be used to monitor low-titre samples from animals at the subclin ical stage. Results became available within 24 h, thus rendering the p rocedure more rapid than ELISA and culture techniques. Six other metho ds designed for milk or other complex samples were tested in this stud y, but found unsatisfactory. Rapid and specific detection of the patho gen by PCR from nasal mucus treated with lysis buffer and proteinase K was demonstrated for swabs taken from experimentally infected calves. Both methods represent useful tools for effective livestock monitorin g and single-animal diagnosis.