Rapid identification of Mycobacterium avium subsp. paratuberculosis laboratory strains by IS900-nested polymerase chain reaction

Publication date: Available online 27 October 2016 Source:International Journal of Mycobacteriology Author(s): M. Mohammad Taheri, N. Mosavari, M.M. Feizabadi, K. Tadayon, R. Keshavarz, R. Aref Pajoohi, K. Soleimani, Sh. Dashti Pour Mycobacterium avium subsp. paratuberculosis (MAP) causes paratuberculosis (Johne’s disease) in ruminants. As a species, M. avium comprises M. avium subsp. hominissuis and a number of clones that are known to have evolved from this subspecies, namely M. avium subsp. avium (MMA), M. avium subsp. silvaticum, and MAP. Despite the very high genomic similarity of MAP and MAA, the insertion sequence IS900, which is 1,451-bp long, is now understood to be exclusively present in 10–20 copies in the genome of MAP. In the present study, a multidiscipline polymerase chain reaction (PCR)-based algorithm targeting16SrRNA, IS6110, IS901, IS1245, and IS900 markers has been employed to differentiate between six laboratory strains of M. avium complex (including MAP 316F, III&V, and 2e plus MAA D4), Mycobacterium tuberculosis DT, and Mycobacterium bovis AN5 strains used at the Razi Institute (Tehran, Iran) for the preparation of paratuberculin, avian, human, and bovine tuberculin, respectively. Three laboratory strains of III&V, 2e, and 316F were subcultured on Herrold’s egg yolk medium, whereas the MAA strain of D4 along with M. bovis AN5 and M. tuberculosis DT were subcultured on Lowenstein–Jensen slopes. All the inoculated culture...
Source: International Journal of Mycobacteriology - Category: Infectious Diseases Source Type: research