This assay is known as the tier-two blood test for the confirmation of Lyme disease with high accuracy. It detects Borrelia-specific peptide antibodies in human serum, was refined by different investigators, and was recommended for a better diagnosis of Lyme disease.
The advantage of Western Blot over the ELISA method is that while ELISA uses a mixture of proteins for testing, in WB, the antigens are separated based on their molecular size, for example, 18, 23, 28, 30, 39,
41, 45, 58, 66, and 93 kDa prior to completion of the ELISA steps. The disadvantage is that the antibody reaction with each pure antigen can only be observed qualitatively or simply a positive or negative, yes or no result.
A sample-positive Western Blot test is shown in the figure below.
The Western Blot (WB) assay has been widely used to detect the presence of antibodies in human serum and plasma to various infectious disease agents.
In this procedure, component proteins of purified, inactivated organisms are electrophoretically separated by SDS-polyacrylamide electrophoresis followed by electrotransfer to nitrocellulose sheets. Each strip serves as the solid-phase antigen for an ELISA test.
Specific antibodies present in human serum/plasma will bind to several of the separated polypeptides upon incubation with the strip. These antibodies are then treated with an enzyme-antibody conjugate which binds to the human immunoglobulins if present. The final product of the membrane antigen, a human antibody, and the conjugate is visualized upon incubation with a chromogenic enzyme substrate. This will result in a blue-colored “band” at the polypeptide location on the membrane strip if the specific human antibody is present.
This panel is beneficial for a wide variety of patients with:
The Borrelia burgdorferi IgG, IgM by Western Blot should be considered when patients present the following symptoms and conditions:
This test confirms the presence of ongoing Lyme disease based on CDC criteria.