PMC2730323_01
Ziehl-Neelsen-stained micrograph of Mycobacterium avium subsp. paratuberculosis colonies growing on mycobactin-supplemented Middlebrook agar.
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PMC2730323_01_01-0388-F1.jpg
F1
Middlebrook slants with colorless dysgonic colonies. Microscopic examination of these colonies showed acid-fast bacilli (Figure 1).
Agarose gel electrophoresis of amplified IS900 fragments. Lanes 1 and 6: molecular weight marker (2176, 1766, 1230, 1033, 653, 517, 453, 394, 298, 234-220, 154 bp); lanes 2 and 3: two patient samples; lane 4: positive control (Mycobacterium avium subsp. paratuberculosis type strain); and lane 5: negative control (Mycobacterium avium strain).
01-0388-F2.jpg
PMC2730323_01_01-0388-F2.jpg
F2
TGTTCGGGGCCGTCGCTTAG; IS900-2: 5 -CGTTCCAGCGCCGAAAGTAT), which is present only in MAP strains, was done with the two most recent positive cultures. This assay showed clearly positive results from the two cultures tested and the MAP type strain, while the M. avium strains remained negative (Figure 2).