Selected MAP binders used to coat MyOne™ tosylactivated Dynabeads® for magnetic separation evaluation.

<p>Selected MAP binders used to coat MyOne™ tosylactivated Dynabeads® for magnetic separation evaluation.</p

Comparison of IMS-PCR and Direct PCR results for 280 lymph node samples –74 non-visibly lesioned (NVL) and 206 visibly lesioned (VL).

a<p>Determined by McNemar's Chi-Square test using Genstat version 14.2.</p>b<p>Includes 166 VL and 74 NVL samples from TB skin test reactor animals plus 40 VL samples from non-reactor animals detected at routine slaughter.</p><p>Data represent number of samples with each combination of test results.</p

Distribution of <i>M. bovis</i> spoligotypes isolated by culture and IMS-MGIT culture.

<p>Spoligotypes 269 (140*), 142* and 263* are thought to be spoligotypes 140, 142 and 263 with spacer 15 signal absent which have arisen due to lesser amounts of <i>M. bovis</i> DNA being present in IMS-MGIT cultures.</p

Schematic overview of the phage display biopanning approach taken to identify MAP-specific peptide binders using irradiated MAP B4 whole cell antigen (WCA) and ethanol extracted antigen (EEA) targets.

<p>Schematic overview of the phage display biopanning approach taken to identify MAP-specific peptide binders using irradiated MAP B4 whole cell antigen (WCA) and ethanol extracted antigen (EEA) targets.</p

Interrelationships between Culture, Direct PCR and IMS-PCR positive results for all 280 lymph node samples, irrespective of type.

<p>Interrelationships between Culture, Direct PCR and IMS-PCR positive results for all 280 lymph node samples, irrespective of type.</p

MAP capture sensitivity of magnetic beads coated with monoclonal antibodies and peptides to capture whole cells of MAP strain TB13-008781 from PBS dilutions containing approx. 10⁴−10 cfu/ml when used (A) individually, (B) as 50:50 bead combinations, or (C) as dually-coated beads.

<p>DNA extracted from the bacterial suspensions before MS was used as a control. The expected IS900 PCR product is 394 bp. Lane MW indicates a 100 bp DNA ladder (TrackIt™, Life Technologies).</p

Comparison of results for alternative detection methods and culture (MGIT broth or Lowenstein-Jensen slope).

a<p>Determined by McNemar's Chi-Square test using GenStat Release 14.2.</p><p>Results for all 280 lymph node samples are not broken down by lymph node type.</p

Outgrowth of milk bacteria non-specifically adhering to four types of coated magnetic beads in Middlebrook 7H9 OADC broth without (■) and with antibiotic supplements (NOA ◆ and PANTA ▼) after MS was applied to raw milk.

<p>Outgrowth of milk bacteria non-specifically adhering to four types of coated magnetic beads in Middlebrook 7H9 OADC broth without (■) and with antibiotic supplements (NOA ◆ and PANTA ▼) after MS was applied to raw milk.</p

Competitive ELISA results obtained for WCA and EEA generated monoclonal antibodies.

<p>Bacterial suspension of irradiated MAP was added wells as ‘free MAP cells’ prior to the addition of the antibody. Negative control wells received assay buffer followed by the antibody. A decrease in absorbance values in the presence of ‘free MAP cells’ over negative control wells is indicative of MAP binding. Figure depicts the mean absorbance values (n = 3) ± standard error. Mean % binding inhibition was calculated based on the difference in the % binding of the antibody to the attached MAP cells in the presence and absence of ‘free MAP cells’, where negative control wells represent 100% binding.</p

Percentage non-specific capture of a range of <i>Mycobacterium</i> spp. (detailed in Materials and Methods) by variously coated Dynabeads.

<p>Figure depicts mean percentage capture ± standard error of results for ten different <i>Mycobacterium</i> spp. captured by each bead or bead combination: MAb 6G11 coated beads; beads coated individually with MAb 6G11 and MAb 15D10 and used as 50:50 mix; beads dually coated with MAb 6G11 and MAb 54.3.1.AA; beads coated individually with biotinylated aMp3 and aMptD peptides and used as 50:50 mix (control).</p