An automated PCR with fluorescent probes (molecular beacons) detected Mycobacterium avium subsp. paratuberculosis in bovine feces. When the PCR was compared with culture in testing 41 fecal samples, kappa scores of 0.94 to 0.96, a sensitivity of 93 to 96%, and a specificity of 92% were obtained. Results were quantitated by using a standard curve derived from a plasmid containing IS900. A minimu...

The aim of this study was to detect the IS900 region of Mycobacterium avium subsp. paratuberculosis (MAP) in bovine milk samples using real-time polymerase chain reaction (qPCR) and conventional PCR, and to study the agreement between these tests. A total of 121 bovine milk samples were collected from herds considered positive for MAP, from the State of Pernambuco, Brazil. MAP DNA was detected ...

The objective of the work was to evaluate two ELISA’s for detection anti-Map antibodies. 460 serum and fecal samples bovines (n=267), ovines (n=140), goats (n=53) were evaluated. Serums used detecting antibodies using different ELISAs, one in-house ELISA made with low-molecular weight proteins (<100 kD) from antigen ultra-filtrate Map strain 3065, a commercial paratuberculosis diagnost...

The aim of this study was to confirm clinical diagnosis of paratuberculosis in two cows showing suggestive clinical signs of the disease. Based on clinical signs, in culture and in IS900 PCR results from the individual milk samples it was possible to diagnose paratuberculosis in the cows studied.

Mycobacterium avium subsp. paratuberculosis is the etiologic agent of Johne's disease and is endemic to the national cattle herds of many countries. Because of the very low level of genetic heterogeneity of this organism, it is difficult to select a workable procedure for strain differentiation at a resolution sufficient to investigate epidemiological links between herds or different ruminant s...

With the objective of identifying presence Paratuberculosis (PTB), an infectious disease caused by Mycobacterium avium ssp paratuberculosis (MAP), in sheep, through pathological studies, bacterial culture and IS900 PCR, as well estimating seroprevalence to MAP. The present cross-sectional study, was conducted 16 different flocks, with serum 2415 adult sheeps, analyzed Enzyme-Linked ImmunoSorben...

Bacterial culture is the 'gold standard' for detecting Mycobacterium avium subspecies paratuberculosis (MAP) infection, but is time consuming, laborious, and recovery of organism varies with species of animal tested. PCR has been used for detection of MAP DNA in feces and tissues. We used PCR to detect MAP DNA isolated from tissues from 25 free-ranging North American bison (Bison bison), each w...

The analysis of the gyrA and gyrB genes of a panel of Mycobacterium avium subsp. paratuberculosis isolates from types I, II, and III detected type-specific single nucleotide polymorphisms. Based on these results, we developed a PCR and restriction enzyme analysis to discriminate type I and III isolates. The application of this technique would be the unique strategy to characterize these strains...

Mycobacterium avium subspecies paratuberculosis (MAP) causes Johne's disease, a chronic progressive enteritis. It is generally assumed that calves rarely shed MAP bacteria and that calf-to-calf transmission is of minor importance. The objectives were 1) to estimate the prevalence of MAP-shedding young stock in MAP-infected dairy herds, and identify predictors for test-positive young stock; and ...

Figure 1. Representative PCR results showing amplification (or otherwise) of 298-bp genomic fragment corresponding to IS900 in diabetic patients (lanes 1–8). Lanes 9 and 10, Control subjects without type 1 diabetes mellitus. Lanes 11 and 12, Mycobacterium avium subspecies paratuberculosis–positive and M. avium subspecies paratuberculosis–negative control subjects, respectively. M, molecular mar...