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Antibody Detection Using Serological Assays
As clinical diagnostic and surveillance tools, serological assays such as the enzyme-linked immunosorbent assay (ELISA) and the lateral flow immunoassay (LFIA) have been extensively used in the detection and identification of viruses and other pathogens. They achieve this aim through the use of highly specific recombinant antigens, or fusion proteins, to detect serum antibodies, which are used by the immune system to neutralize pathogenic bacteria, viruses or other microorganisms that cause disease. Serological assays have been successfully used in the detection of hepatitis viruses, the Ebola virus, HIV and influenza.
Serological tests that have been rigorously validated for accuracy have the potential to transition from the laboratory setting to the point-of-care. Point-of-care-testing is a medical diagnostic test administered at or near the patient, such as a medical facility. (It is important to note that AAT Bioquest provides reagents intended for research use only.)
Representation of direct and indirect antigen detection using target-specific antibodies.
ELISA, or enzyme-linked immunosorbent assay, is a plate-based immunoassay used to detect and quantify biomolecules such as antibodies, proteins, hormones or peptides, as well as for the characterization of protein-protein and protein-nucleic acid interactions. In an ELISA, the target of interest, typically an antigen, is immobilized onto a solid surface and subsequently probed with an antibody conjugated to an enzyme such as horseradish peroxidase (HRP) or alkaline phosphatase (AP).
Secondary antibodies are key components of the indirect detection method. Used in sequence with primary antibodies, secondary antibodies facilitate in the detection, quantification and purification of target antigens by binding to the primary antibody, which directly binds to the antigen. Reporter molecules (e.g. an enzyme or a fluorophore) conjugated to the secondary antibodies enables visualization of the immune complex.