What is Elisa's detection?

The enzyme connected by the immunosorbent test (ELISA) is a test performed in the immunological laboratory for determining protein levels in the biological sample. The ELISA detection concerns the last step of the test in which a clear solution or substrate is added to the plastic plate containing a bound antibody marked enzyme. The enzyme cleaves the substrate and the color changes. The absorbance of the final color solution is then measured on the ELISA reader or spectrophotometer. The indirect Elisa is used to detect a protein known as an antibody, in the patient's serum. An example of the indirect Elisa test is the HUV detection (HIV antibodies detection. The ELISA sandwich test detects protein or antigen, capturing between two antibodies. The detection of the hormone of human chorionic gonadotropin (HCG), which is Evybirán during pregnancy, is performed with the ELISA sandwich test.

both tests have a detection at the end of the testElisa's step. This step involves adding an antibody that has the enzyme molecule attached to it. After adding an enzyme -marked antibody, a colorless solution containing a substrate -specific molecule is added. The enzyme cleaves the substrate molecule and the solution changes color depending on the combination used. Determination of the amount of antibody or antigen in the patient sample is measured by measuring the intensity of color change.

There are several combinations of enzyme substrates for use in the ELISA detection step. The most common enzyme is the horseradish peroxidase (HRP), which may, among other things, break down the molecules of ortho-phenylemendiamine dihydrochloride (OPD) and tetramethylbenzidine (TMB). The cleavage of both OPDA TMB results in yellow color and the optical density or absorbance of the light of these substrates is measured by the ELISA reader. OPD absorbance is measured at a wavelength of 490 nanometers (Nm), while TMB is measured at 450 Nm.

Another common enzyme used in the detection step of ELISA is alkaline phosphatase. This enzymeIt is used with a substrate by p-nitrophylphosphate (PNPP) and also produces a yellow solution. PNPP absorbs light on a wavelength of 405 nm.

The selection of the regions of the enzyme substrate is usually based on what antibodies marked with the enzyme are commercially available and also what devices will be used to measure light absorbance. Many combinations available for ELISA detection make the Elisa test very versatile. It is an important tool in testing diseases and research laboratories.