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Applied Immunology: Enzyme-Linked Immunosorbent and Enzyme Linked Immunospot Assays - Research Paper Example

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The author describes the enzyme-linked immunosorbent assay, a highly sensitive method employed to detect the presence of antigens in a variety of samples, and the Enzyme-Linked Immunospot assay which measures the frequency of cytokine-secreting cells at a single cell level…
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Applied Immunology: Enzyme-Linked Immunosorbent and Enzyme Linked Immunospot Assays
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Download file to see previous pages First introduced in the early 1970s, the enzyme-linked immunosorbent assay is a highly sensitive method employed to detect the presence of antigens in a variety of samples. The technique, which uses an enzyme as a label, involves binding antigen to the solid phase, incubating then wash in the ELISA plate to clear unbound excess antigen. The diluted serum suspected to be containing specific immunoglobulin antibody for bound antigen is added to the ELISA plate/ well and allowed time to react. Enzyme-labeled immunoglobulin is then added, allowed time for attachment then wells washed. The substrate of the enzyme-labeled antibody is then added. The enzyme hydrolyzes or breaks down the substrate causing color production. The optical density of the final color is directly proportional to the quantity or amount of unknown antibody or antigen present. (NOTE: Same procedure is repeated for the detection of antigen in a serum) (Bach, 1982, 58) ELISA is mainly applied in immunodiagnosis of infectious disease including measurement of viral antigens such as herpes, coxsackie and so on, measurement of bacterial and mycotic antigen such as brucella antigen, salmonella antigen and aflatoxin antigen, measurement of antigens to Trypanosoma cruzi and Schistosoma mansoni and the measurement of antibodies to viruses such as rubella, measles, herpes, rabies and so on. Various components in the blood sample of non-infected individuals can also be measured through ELISA, for example, hormone-insulin (Guttmann, 1981, 365). Some of the advantages of the Enzyme-linked immunosorbent assay include the fact that it does not require a radioactive substance/ radioisotope or a costly radiation counter (radiation counting apparatus). It is also advantageous in the sense that it is a highly sensitive and specific method and hence relatively very accurate. However, there are a few disadvantages relating to its application in immunology.  ...Download file to see next pages Read More
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