Presence, characterization, and genotype profiles of Mycobacterium avium subspecies paratuberculosis from unpasteurized individual and pooled milk, commercial pasteurized milk, and milk products in India by culture, PCR, and PCR-REA methods

Agarose gel with the IS900 specific primers, which generate a fragment that migrates at a molecular weight of 229bp. Lane 1=markers; lanes 2–10 are all samples that had been grown in culture, had had DNA extracted, and were now tested for the presence of IS900-specific MAP DNA sequences. All samples shown here tested positive for the presence of IS900.

Agarose gel with the IS900 specific primers, which generate a fragment that migrates at a molecular weight of 229bp. Lane 1=markers; lane 2 is the positive MAP DNA control; lane 3 is the negative control (water processed identically with the other samples); lanes 4–10 are all samples that had been isolated from the pellet of samples in this study. All samples show a positive signal.

Differentiating MAP into ‘bison type’ and ‘cattle type’ using HinfI and MseI restriction endonuclease analysis (REA) and IS1311 PCR. Shown is a 4% agarose gel of digested DNA subjected to MAP-specific IS1311 PCR. Lane 1=molecular weight markers; ‘cattle type’ MAP results in three fragments (lanes 2, 3, and 5);⁴³ in contrast, ‘bison type’ MAP generates only the two larger bands (lanes 4, 6, 7, and 8).⁴³.