Detection of quinolones resistance (QRDR) genes
Amplification of genes (gyr A and par C) encoding for
quinolone resistance (QRDRs) was carried out by PCR. The PCR reactions
were performed in 50 μl volume for each isolate, with 30 pmol/μl of each
primer and 100 ng DNA. Conditions for the PCR were as follows: 95°C for
3 min, 30 cycles of denaturation for 30 sec at 95°C, followed by
annealing of 30 sec at 56oC and extension at 72°C for
45 sec with a final extension at 72°C for 10 min. The type strainM. bovis ATCC 25523/ PG45 was used as quality control