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Question

The key reagent used in ELISA is

A
catalase
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B
DNA probe
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C
alkaline phosphatase
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D
RNase
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Solution

The correct option is C alkaline phosphatase
ELISA is the basic assay technique, known as Enzyme Linked Immuno-sorbent Assay.

It is rapid, quick and requires a blood sample of the patient. ELISA is used to detect and quantify substances, including antibodies, antigens, proteins, glycoproteins, and hormones. The detection of these substances is accomplished by complexing antibodies and antigens to produce a measurable result.

The entire procedure of ELISA is as follows:

An antibody (referred to as primary antibody) is attached to a plate/solid surface that has an affinity towards pathogens.

Blood samples containing antigen are added, after which free antibodies are removed by washing.

A secondary antibody which can bind to the antigen is added. Secondary antibodies are usually conjugated with enzymes like alkaline phosphatase. Alkaline phosphatase is commonly used enzyme and is a key reagent to perform the ELISA test.
Free enzyme-linked secondary antibodies are removed by washing the plate.

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

The amount of reflection, absorption and the colour in the spectrophotometer identifies and measures the amount of antigen/antibody present in the blood.


Hence, ELISA testing is based on the principle of antigen-antibody interaction.
Infection by pathogen can be detected by the presence of antigens or by detecting the antibodies synthesised against the pathogen.

Catalase is an enzyme that breaks down harmful hydrogen peroxide into oxygen and water. It is present in the liver and is usually not used in the ELISA test.

DNA probes are single stranded fragments of DNA that contain a nucleotide sequence complementary to the target sequence (sequence to be detected). They are usually labelled with radioisotopes to enable their detection.

RNase or ribonuclease is a type of nuclease that catalyses the breakdown of RNA into smaller components.

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