They are used to determine whether pathogen-specific antibodies are present in patient serum or plasma. The IgG and IgM Western Blots are qualitative immunoassays in which antibodies are visualized. Each spot that develops in the assay represents a single reactive cell. The IgXSpot assay allows visualization of the secretory product(s) of individual activated or responding cells to borrelia-specific antigens. Therefore, the IgXSpot test is recommended for detection of very early and/or late borrelia/pathogen infection and in seronegative patient’s whole blood samples.ĮLISPOT is a widely used method for detecting and monitoring cellular immune responses to specific antigens. In some patients with a chronic form of the disease, the humoral response is poor. In some patients, sero-conversion from cellular to humoral response does not occur or occurs much later in the disease. ![]() The cellular immune response develops much earlier than the humoral response in most patients who are infected with borrelia species. It is well documented that both humoral and cellular immune responses develop in borrelia/pathogen infection. The IgXSpot is an enzyme-linked immunospot assay that detects human T cells reactive to borrelia/pathogen-specific antigens in vitro. ![]() Bands are visualized and scored for intensities relative to the positive and negative controls. A dark purple colored precipitate develops on the antigen-antibody complexes. Bound antibodies react with BCIP/NBT, a chromogenic substrate. Step 3: After washing off the unbound serum, the strip is incubated with alkaline phosphate-conjugated goat anti-human antibody. If specific antibodies to pathogen antigens are present, they will bind to the corresponding antigen bands. Step 2: Patient serum or plasma is incubated with the IB strip. The recombinant proteins are applied to a membrane and cut into strips. ![]() Step 1: IB strip preparation: IB strips are prepared from recombinant proteins of interest. These tests are generally more sensitive and specific than the Western Blot, ELISA, and IFA tests. The IgM and IgG ImmunoBlot (IB) are qualitative immunoassays in which antibodies are visualized.
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