Michael Bovee
The enzyme-linked immunosorbent assay is a widely used analytical chemical assay pioneered by Engwall and Perlman in 1971. The assay includes a non-gel solid phase antibody immunoassay that detects the presence of antigen in an inorganic -soluble or insoluble solution using specific antibodies guided by an antigen binding strategy.
The elISA test can be performed on a variety of biological samples, including: human blood, human saliva, mouse blood, mouse serum, and animal serum. The assay was originally developed to measure antigen levels in blood samples, but its sensitivity has also increased dramatically. Today it is widely used to measure the level of antibodies in various biological fluids. It is a very accurate and sensitive method for detecting tiny amounts of antigen in biological fluids. This precision makes elISA an important tool in the diagnosis of various diseases and health conditions.
The elISA test is based on the assumption that antibodies bind to an antigen. Antibodies are produced by different types of cells in the immune system. They usually attack pathogenic microorganisms present in the body, but some of them are designed specifically to attack pathogenic antigens and cause allergies or even chronic diseases. Elisa uses antibodies to discriminate between pathogenic and non-pathogenic antigens for reliable results.
As previously mentioned, elISA can be performed on various types of biological samples, including: human blood, human saliva, mouse blood, mouse serum, and animal serum. ELISA can also be performed using a variety of solvents including water, acetonitrile, hexamethylenetetramine, and dimethylformamide.
The ELISA testing procedure consists of several stages. The first step involves preparing a sample solution that includes an antigen binding agent, an indicator antigen, and a buffer.
After preparation of the samples, the sample solution is tested using the elISA test on a laboratory plate by exposing it to a dye in the absence of antigen and antibody. The plate reader analyzes the color and concentration of the dye to determine the concentration of antigen and antibodies bound to the plate. The second step involves obtaining an electrophoretic mobility shift curve, which is obtained from a test plate, to confirm the antigen binding relationship between the sample and the dye.
The third step involves checking the ELISA test results by observing the change in plate displacement after the dye has been replaced with water. The fourth step involves preparing a reference ELISA plate to compare the dye reactive stain on the sample to the reference plate. The fifth step involves preparing a control plate for comparison purposes as well as an elastomeric control for the gel used for the elisabet test. Finally, the gel is incubated in a buffer containing a small amount of antigen binding agent in an incubator at room temperature for up to 2 hours to ensure that the gel solidifies well and allows the antigen binding agent to react with the dye.
As mentioned earlier, an ELISA test is used to detect the specific binding of antibodies to an antigen. Therefore, it can only detect antigens that are in the same solution as the antigens tested. Moreover, it cannot detect small amounts of other molecules. in the sample. However, it can detect other molecules such as sugars, lipids, glycerol and triglycerides, as well as certain proteins that are not present in the antigen binding solution.
An ELISA test is usually performed using a plate reader to read the result. The use of a tablet reader ensures high sensitivity of the test.
Some patients have been treated with the elisa test, but there is still debate about its safety and effectiveness. Some of these patients have some degree of hemolysis. The most common treatment is intravenous heparin or intravenous cannula infusion, which is given after the patient is admitted to the hospital for hemolytic anemia.
The elisa test is widely used in the United States for clinical purposes in the fields of gastroenterology and cardiology. This test is used in the USA instead of the LFT test and is sometimes combined with the ETS test. in the case of patients with mild anemia. In addition, it is used in the field of the cardiovascular system to determine the effect of drugs on the concentration and function of red blood cells.