



| Name | Mitochondria ELISA test kit |
|---|---|
| Category Name | Autoimmune Disease kits |
| Test | 96 |
| Method | ELISA method: Enzyme Linked Immunosorbent Assay |
| Principle | ELISA principle - Peroxidase conjugated |
| Detection Range | Qualitative elisa assay: Positive, low positive, negative |
| Sample | 10ul |
| Specificity | 100% |
| Sensitivity | 100% |
| Total Time | ~60 min |
| Shelf Life | 12 months |
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The Diagnostic Automation Mitochondria Enzyme- Linked Immunosorbent Assay (ELISA) is intended for the detection and semi-quantitation of antibodies to mitochondria in human sera. The assay is to be used to detect antibodies in a single serum specimen. The results of the assay are to be used as an aid to the diagnosis of primary biliary cirrhosis.
Systemic autoimmune disease is characterized by the presence of circulating auto-antibodies directed to a wide variety of cellular antigens. Systemic lupus erythematosis (SLE) commonly referred to as Lupus is the best known of these diseases. Other possible connective tissue diseases include mixed connective tissue disease (MCTD), Sjogren syndrome, scleroderma, and polymyositis/dermatomyositis. The majority can be diagnosed by clinical presentation and their antibody profiles to the various antigens involved, which include dsDNA, Sm, RNP, Ro, La, Scl-70, Jo-1 and Histones. Therefore, immunoassays for autoantibody are useful for diagnostic and prognostic evaluations of autoimmune disease. Mitochondria (M2) antigen is part of the pyruvate dehydrogenase complex. Approximately 90-95% of patients with primary biliary cirrhosis (PBC) have antibodies to M2. Antimitochondria antibodies occur occasionally in other liver conditions and scleroderma. The antibody is rarely seen in other conditions. Classically, antibodies to autoimmune antigens are detected by double immunodiffusion. However, the test is lengthy and suffers from weak sensitivity. Enzyme-Linked Immunosorbent Assays (ELISAs) combine greater sensitivity with ease of use. Many ELISAs have been developed and validated for detecting autoantibody to various antigens.
The DAI Mitochondria test is an Enzyme-Linked Immunosorbent Assay to detect IgG, IgA, and IgM antibodies to Mitochondria antigens. Purified Mitochondria antigens are attached to a solid phase microassay well. Enzyme-Linked Immunosorbent Assays (ELISA) relies on the ability of biological materials (i.e., antigens) to adsorb to plastic surfaces such as polystyrene (solid phase). When antigens bound to the solid phase are brought into contact with a patient's serum, antigen specific antibody, if present, will bind to the antigen on the solid phase forming antigen- antibody complexes. Excess antibody is removed by washing. This is followed by the addition of goat anti-human IgG, A, M conjugated with horseradish peroxidase which then binds to the antibody-antigen complexes. The excess conjugate is removed by washing, followed by the addition of Chromogen/Substrate, tetramethylbenzidine (TMB). If specific antibody to the antigen is present in the patient's serum, a blue color develops. When the enzymatic reaction is stopped with 1N H2SO4, the contents of the wells turn yellow. The color, which is indicative of the concentration of antibody in the serum, can be read on a suitable spectrophotometer or ELISA microwell plate reader.

