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Title: Dot-immunobinding assay.
Authors: Sumi, S
Mathai, A
Radhakrishnan, V V
Keywords: Immunology
Issue Date: 2009
Publisher: Methods in molecular biology (Clifton, N.J.)
Citation: Methods in molecular biology (Clifton, N.J.). 536; ; 89-93
Abstract: Dot-immunobinding assay (Dot-Iba) is a simple and highly reproducible immunodiagnostic method. Antibody or antigen is dotted directly onto nitrocellulose membrane (NCM) discs. The diagnostic material to be checked can be incubated on this disc. Presence of antigen-antibody complex in NCM discs can be directly demonstrated with enzyme-conjugated antiglobulins and substrate. Development of a purple-pink colored, insoluble substrate product in the NCM will be considered a positive result in the assay. This assay allows the processing of multiple specimens at a time and the entire operational procedures require only 4-6 h. Dot-Iba is rapid, and the technical steps involved in the assay are much simpler than in the other immunoassays such as enzyme-linked immunosorbant assay in detecting circulating antigen and antibody in clinical samples. The Dot-Iba showed an overall sensitivity of 60% for tuberculous meningitis diagnosis and no false positive results were encountered. Hence, this assay is highly specific for the diagnosis of paucibacillary diseases such as extrapulmonary tuberculosis. Dot-Iba is best suited to laboratories in developing world where there are constraints in laboratory resources.
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