LIVER DISEASE

Instructions for use

The MESACUP LKM-1 TEST is semi-quantitative enzyme-linked immunosorbent assay (ELISA) for the detection of anti-LKM-1 antibodies in human serum.

Background:

The causes of the chronic liver disease include viruses (HBV, HCV etc.), alcohol, metabolic errors, and others. An autoimmunity plays a definite role in pathogenesis of a group of chronic active hepatitis as auto-immune hepatitis (AIH). One of the main characteristics of AIH is the presence of autoantibodies such as anti-nuclear antibodies (ANA), anti-smooth muscle antibodies (ASMA) and others. Rizzetto et al. in 1974 reported a new type of autoantibody in patients with chronic active hepatitis which selectively reacts with proximal tubular epithelia of rat kidney sections with different pattern from anti-mitochondrial antibodies (AMA) by indirect immunofluorescence antibody method (IIF). This autoantibody was studied in detail by Homberg et al. and was named as anti-Liver / Kidney microsome (LKM) antibodies. Manns et al. characterized a subgroup of AIH type II by the presence of the antibodies and this clinical feature. Anti-LKM antibodies react with the tissues sections of both liver and kidney by IIF, and were classified into 3 subtypes by staining patterns; anti-LKM-1 antibodies are detected in type II AIH, anti-LKM-2 in tienilic acid-induced hepatitis and anti-LKM-3 in chronic hepatitis D. Alvarez et al. in 1985, reported that anti-LKM-1 antibody in patients sera recognize a 50KD protein in rat liver microsomes and the protein has been identified as cytochrome P450IID6 in human liver by Manns and others. At present anti-LKM antibody is examined by the IIF using rat kidney as a substrate. However, the technique needs some skillfulness in judgment especially when autoantibodies to ANA and AMA coexist.

The MESACUP LKM-1 TEST is for measuring anti-LKM-1 antibodies present in the serum with high sensitivity by ELISA.

Principle:

The MESACUP LKM-1 TEST measures anti-LKM-1 antibodies present in the serum by ELISA. Calibrators and patient serum are added to microwell coated with LKM-1 antigens, allowing anti-LKM-1 antibodies to react with the immobilized antigen (Sample incubation). After wash to remove any unbound serum proteins, horseradish peroxidase conjugated anti human IgG and IgM are added and incubated (Conjugate incubation). Following another washing step, the peroxidase substrate is added and incubated for an additional period of time (Substrate incubation). Acid solution is then added to each well to terminate the enzyme reaction and to stabilize the color development. The assay can be quantified by measuring the reaction photometrically and plotting the results.

Size: 96 wells

Method: ELISA

Substrate: Recombinant