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ELISA can be defined as a polystyrene plate-based assay process intended for investigating and accounting for protein elements, peptides, hormones, and antibodies. An ELISA process is used to detect an antigen and that antigen should be static to a hard surface and then rebounded with an antibody to associate with an enzyme. ELISA full form refers to an Enzyme-linked immunoassay. ELISA test is used for ascertaining the antigen present in the blood cell. This process is finished in the process of compounded enzyme activity due to incubation. ELISAs are fundamentally executed in 96-well or 384-well plates of polystyrene, those are active to connect or bind proteins and antibody elements. This binding process and immobilization of compounds lead the ELISA process to accomplish its purpose and to detect antigen type.
Define ELISA or enzyme-linked immunoassay
An enzyme-linked immunosorbent assay, also called ELISA or EIA, is an experiment that discovers and evaluates antibodies and antigens in the blood. This test is effective to determine if a patient has specific antibody-related infections. The patient’s body generates in reaction to a harmful protein substance named antigens. In 1972, Eva Engvall and Peter Pearlman first invented the immunoassay process that greatly impacted immunology and medical science. An ELISA test is effective to diagnose diseases such as AIDS, pernicious anemia, Lyme disease, rotavirus, syphilis, Rocky Mountain spotted fever, toxoplasmosis, varicella-zoster virus, which gave rise to chickenpox, etc.
Principles of ELISA
ELISAs are generally implemented in plates of 96-well polystyrene. The incubation process has begun with the serum being stored well. Although every well of the assay consists of a variety of serum. There must be 96 specimens being investigated including the positive control and negative control.
The serum consists of various kinds of antigen and antibodies and is apprehended by correlating antigen or antibody enclosed onto rigid surfaces. With some passing time, the plate is cleaned to pull out the serum and unchain antibodies or antigen with a sequence of wash intermediaries.
To investigate the chained antibodies or antigen, subordinate antibodies that are secure to an enzyme for instance peroxidase or phosphatase are appended to every well. The unbound subordinate antibodies are washed away following the incubation timespan.
According to the ELISA principle, a color variation has been noticed at the time a suitable component is added up. This change of color will indicate a tool to investigate the function or accountability of the antigens. The potency of the color gives a prospectus of the quantity of antigen or antibody. The density of color is calculated at 450 nm to investigate the quality of the antigen
Limitation of ELISA Test
- The main limitation is yielding false results. The intensity of color strength will demonstrate the irregular investigation about the presence of antibodies in a specific specimen.
- Another limitation is insufficient scalability. Fundamentally performing ELISA within 96-well plated format, ELISA is unable to scale through to increase the miniaturization thoroughly.
- Instead of an automated plate washer, ELISA tends to be a tedious assay with numerous washing steps. The washing step is significant to generate a linear standard curve and an inadequate washing step will hamper the step.
Difference between Immuno-PCR and ELISA test
Immuno-PCR is an investigating tool with an ascending signal improved version of PCR. In which the PCR is utilized with a DNA marker to detect the targeted antigen-antibody. Both technologies have the same objective of detecting the antigen. But ELISA is detecting antigen in presence of enzyme whereas Immuno-PCR has utilized the DNA to accomplish the detection. Along with that Immuno -PCR obtained more sensibility than ELISA.
Conclusion
Therefore, this paper concludes with a brief discussion against the ELISA test or EIA. The main purpose of the ELISA is to detect the antigen in a specimen. This procedure helps to investigate many infections occurring from plants to humans or animals. ELISA Principle is that particular antibodies attach with the aimed antigen and dig out the existence and quality of the antigen. Especially disease or infection with blood borne diseases including HIV, HCV, and HTLV, for screening those diseases ELISA test is implemented in general. Hence, the ELISA test is used for early detection of the antigen quality and nature of the infection. Collective and transportable ELISA is accessible and a reduced cost lab kit is also significant for large amounts of population screening in lesser resource arrangements.
