Immunochromatographic flow-injection method for detection of human serum immunoglobulin G antibodies to Helicobacter pylori |
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Authors: | D Narinesingh TT Ngo |
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Institution: | a Faculty of Agriculture and Natural Sciences, Department of Chemistry, University of the West Indies, St. Augustine, Trinidad and Tobago b Sino-American Medical Inc., Tustin, CA, USA |
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Abstract: | This study reports on the development of an immunochromatographic flow-injection (FI) method for the quantitation of human serum IgG antibodies to Helicobacter pylori. Patients’ sera were injected into the carrier stream of a FI manifold incorporating an H. pylori immuno-adsorbent reactor. The immuno-adsorbent was prepared by covalently linking H. pylori antigens to periodate oxidized Sepharose CL-4B. Any H. pylori antibody present in the serum is bound to the immuno-sorbent. The bound antibody was quantified by injecting into the carrier stream anti-human IgG peroxidase conjugate followed by TMB/H2O2 as the enzyme substrate. The FI system was optimized with respect to antigen loading (1.0 mg/ml gel), bioreactor length (3.0 cm), enzyme conjugate (0.2 mg/ml) and sample loop size (250 μl). One hundred clinical samples were analyzed for their H. pylori antibody concentrations and the results evaluated with respect to their receiver-operator characteristics (ROC). When a cut-off absorbance of between 0.7 and 0.75 was used for positive and negative samples a specificity (>95%), a sensitivity (>90%)and an overall accuracy (>94%) were obtained. |
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Keywords: | H pylori antibodies Immunochromatography Flow-injection analysis |
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