#ELISA #elisa_in_hindi #antigen_antibody_reaction_in_hindi
#Enzyme_linked_Immunosorbent_assay
Akash gour
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What Is ELISA?
ELISA is the basic assay technique, known as enzyme-linked immunosorbent assay (also referred to as EIA: Enzyme Immunoassay) that is carried out to detect and measure antibodies, hormones, peptides and proteins in the blood.

Antibodies are blood proteins produced in response to a specific antigen. It helps to examine the presence of antibodies in the body, in case of certain infectious diseases.

ELISA is a distinguished analysis compared to other antibody-assays as it yields quantitative results and separation of non-specific and specific interactions that take place through serial binding to solid surfaces, which is normally a polystyrene multiwell plate.

Types Of ELISA
ELISA tests can be classified into three types depending upon the different methods used for binding between antigen and antibodies, namely:

Indirect ELISA – Antigen is coated to the microtiter well

Sandwich ELISA – Antibody is coated on the microtiter well

Competitive ELISA – Microtiter well which is antigen-coated is filled with the antigen-antibody mixture.

Indirect ELISA
Indirect ELISA detects the presence of an antibody in a sample.

The antigen is attached to the wells of the microtitre plate.

A sample containing the antibodies is added to the antigen-coated wells for binding with the antigen.

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All the free secondary antibodies are washed away. A specific substrate is added which gives a coloured product.

The absorbance of the coloured product is measured by spectrophotometry.

Sandwich ELISA
Sandwich ELISA helps to detect the presence of antigen in a sample.

The microtitre well is coated by the antibody.

The sample containing the antigen is added to the well and washed to remove free antigens.

Then an enzyme-linked secondary antibody, which binds to another epitope on the antigen is added. The well is washed to remove any free secondary antibodies.

The enzyme-specific substrate is added to the plate to form a coloured product, which can be measured.

Competitive ELISA
Competitive ELISA helps to detect antigen concentration in a sample.

The microtitre wells are coated with the antigen.
Antibodies are incubated in a solution having the antigen.

The solution of the antigen-antibody complex is added to the microtitre wells. The well is then washed to remove any unbound antibodies.

antibodies present in the well.

The concentration is then determined by spectrophotometry.

Also Read: Antigens and Immunology

Principle of ELISA
ELISA works on the principle that specific antibodies bind the target antigen and detect the presence and quantity of antigens binding. In order to increase the sensitivity and precision of the assay, the plate must be coated with antibodies with high affinity. ELISA can provide a useful measurement of antigen-antibody concentration.

ELISA Procedure
ELISA is one of the easiest blood tests that can be carried out. It is rapid, quick and requires a blood sample of the patient. The entire procedure of ELISA is mentioned below.

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Finally, the substrate is added. The substrate is converted by the enzyme to form a coloured product, which can be

Diseases That Can Be Diagnosed Using ELISA
ELISA can be used to detect some of these conditions:

Ebola

Pernicious anaemia

AIDS

Rotavirus

Lyme disease

Syphilis

Toxoplasmosis

Zika virus

Carcinoma of the epithelial cells

Advantages Of ELISA
Following are some of the advantages of the ELISA technique:

Results fetched from ELISA gives an accurate diagnosis of a particular disease since two antibodies are used.

Applications of ELISA
The applications of ELISA are discussed below:

The presence of antibodies and antigens in a sample can be determined