Performance of SS-NIE-1 ELISA and Luminex.

<p>Note: * CI = confidence interval</p><p>Performance of SS-NIE-1 ELISA and Luminex.</p

Cross-reactivity of SS-NIE-1 ELISA and Luminex.

<p>NT = Not tested</p><p>Cross-reactivity of SS-NIE-1 ELISA and Luminex.</p

Box and whiskers plot of seven groups of sera tested for anti-NIE IgG antibodies.

<p>The plots summarize the results of gold standard <i>Strongyloides</i> samples (n = 54) and six control groups comprised of healthy individuals (n = 7) as well as those with trichinosis (n = 8), filariasis (n = 9), schistosomiasis (n = 9), echinococcosis (n = 6) and amebiasis (n = 8). The lower and upper boxes represent the samples in the second and third quartile respectively while the error bars above and below the box correspond to the 95^th and 5^th percentiles. The horizontal lines separating the boxes represent the median and the diamond denotes the mean. X represents the minimum and maximum values. A significant difference was observed between the gold standard <i>Strongyloides</i> and all control groups (<i>p</i><0.001).</p

Schematic representation of a dotLab biosensor.

<p>(A) Each sensor consists of a contiguous array of 8 assay locations spotted on the bottom of a 10 µL flow channel where reagents and samples are introduced. Each assay location is comprised of a repeating pattern of capture molecules arranged in a defined series of parallel lines creating a diffraction grating. (B) Illumination of an assay spot with a laser generates a predictable diffraction image. The intensity of the diffraction image is monitored in real time by a photodiode detector. (C) Increases in the height (<i>h</i>) of the diffraction grating due to molecular binding events results in a proportionate increase in the diffraction image intensity (Δ<i>DI</i>).</p

Serum dilution optimization.

<p>A series of different dilutions of <i>Strongyloides</i> positive serum were analyzed to determine the optimal serum concentration for use in dot-serology assays. Dilutions of 1∶10 and 1∶20 generated the highest antibody signal with no differences between the two dilutions (<i>p</i> = 0.17), while a significant decrease in signal intensity was observed between 1∶20 and 1∶50 dilutions (<i>p</i><0.001). A serum dilution of 1∶20 was determined to be optimal for the dot-based <i>Strongyloides</i> assay. Data represent mean ± SD.</p

Specificity of the Toxocariasis Luminex assay.

<p>Specificity of the Toxocariasis Luminex assay.</p

Performance of Tc-CTL-1 and Tc-TES-26 Luminex based assays.

<p>Note</p><p>* = 95% Confidence Interval</p><p>Performance of Tc-CTL-1 and Tc-TES-26 Luminex based assays.</p

Representative trace of a dot-based serological analysis of a <i>Strongyloides</i> positive serum sample.

<p>The dotLab mX System outputs a real time trace displaying each reagent incubation and wash step in the assay. Note the binding curves representing the immobilization of NIE@D conjugate and serum anti-NIE antibodies. Significant signal amplification is achieved using anti-human IgG antibody conjugated gold nanoparticles (GNP). The three superimposed traces represent the results of a single assay performed with three spot monitoring.</p

Identified proteins from mass spectrometry analysis.

<p>Note</p><p>+ = Molecular Weight</p><p>* = Sequence Coverage</p><p>Identified proteins from mass spectrometry analysis.</p

2-D gel electrophoresis, silver staining and western blotting of <i>Toxocara canis</i> Excretory Secretory Antigens (TES-Ag).

<p>The TES-Ag sample was separated and analyzed using 2D gel electrophoresis and western blotting. Three of the 2DE gels were transferred to nitrocellulose membranes and probed with a strong EIA positive <i>Toxocara</i> human sera pool (A), a negative human serum sample (B), and <i>Baylisascaris procyonis</i> positive serum (C).A reference gel was stained using silver stain (D). The circled spots in D represent proteins that were excised and subjected to mass spectrometry analysis.</p