Western Blot Test
These tests are used to detect specific proteins in a sample.
Western blot test. It is based on the principle of immunochromatography where proteins are separated into polyacrylamide gel according to their molecular weight. A Western blot test is typically used to confirm a positive HIV diagnosis. The Western blot test, also called immunoblotting, is a test for a specific protein within a protein mixture.
In brief, the sample undergoes protein denaturation, followed by gel electrophoresis. A synthetic or animal-derived antibody is created that recognises and binds to a specific target protein. Western blot can be performed via venipuncture or by spinal tap (lumbar puncture) to test the cerebrospinal fluid.
Western blots tests are also known as protein immunoblots. Then that grid is probed with antibodies that react to the specific proteins that are being searched for. Results from a Western blot test are usually reported in terms of titers which is a measure of the amount of dilution a sample needs before no antibodies remain detectable.
The Western blot test is performed after gel-electrophoresis or an enzyme-linked immunosorbent assay (ELISA) test, and it uses antibodies to identify specific proteins. The electrophoresis membrane is washed in a solution containing the primary antibody, before excess antibody is washed. The western blot, or western blotting, is a widely used analytical technique in molecular biology and immunogenetics to detect specific proteins in a sample of tissue homogenate or extract.
Western blotting (protein blotting or immunoblotting) is a rapid and sensitive assay for detection and characterization of proteins. In case of a positive result from this test, the ELISA test was previously followed by a test called a Western blot to confirm the diagnosis. During the test, a small sample of blood is taken and it is used to detect HIV antibodies, not the HIV virus itself.
A high titer means, therefore, a high level of antibodies and is likely to indicate a major response by the body to a current or recent infection.
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