Autoantibodies against Liver Antigens
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Indirect Immunofluorescence Test: EUROPLUS™ Rat Kidney and M2-3E BIOCHIPs
- Rat kidney and M2-3E BIOCHIP: antibodies against mitochondria (AMA).
- Screening test for the detection of antibodies against mitochondria (AMA) including simultaneous confirmation of the subtype AMA M2.
- Indication: primary biliary cirrhosis (PBC).
- Initial dilution 1 : 100; polyvalent conjugate anti-human IgAGM, FITC-labelled.
- Rat kidney is the standard substrate for detecting anti-mitochondrial antibodies. Nine AMA types (M1 to M9) can be differentiated.
- The BIOCHIP coated with M2-3E permits monospecific confirmation of antibodies against the native pyruvate dehydrogenase complex and the recombinant M2 fusion protein (BPO) in one single test procedure, thus a PBC can be diagnosed serologically with confidence.
- This BIOCHIP Mosaic™ can be supplemented as required using additional substrates, e.g. HEp-2 cells (ANA, nuclear dots), rat liver (liver-kidney microsomes, LKM) or rat stomach (ASMA).
Indirect Immunofluorescence Test: BIOCHIP Mosaic™ Rat Liver/Rat Kidney (Liver Mosaic 1)
- Rat liver and rat kidney: antibodies against liver-kidney microsomes (anti-LKM).
- Specific detection of antibodies against liver-kidney microsomes (anti-LKM).
- Indications: autoimmune hepatitis, often associated with extrahepatic syndromes such as arthralgias, glomerulonephritis, vitiligo and chronic inflammatory bowel diseases.
- Initial dilution 1 : 100; polyvalent conjugate anti-human IgAGM, FITC-labelled.
- Autoantibodies against liver-kidney microsomes react with rat liver and generate a smooth staining in the cytoplasm of the hepatocytes.
- In rat kidney, particularly in the cortex area, a fine granular fluorescence of the proximal tubules - recognizable by the luminal brush border - is visible. The distal tubules are negative. The fluorescence intensity of the liver cells is normally stronger than that of the proximal renal tubules.
- The differentiation between autoimmune hepatitis and virus-induced hepatitis can additionally be accomplished by investigating the appropriate viral parameters.
Indirect Immunofluorescence Test: Liver Mosaic 8
- HEp-2 cells: anti-nuclear dots. Primate liver: anti-LSP. Rat kidney: AMA. Rat liver: ANA. Rat stomach: ASMA. VSM47: Anti-actin.
- Screening and differentiation test for the detection of liver-specific antibodies, antibodies against mitochondria (AMA), antibodies against cell nuclei (ANA), antibodies against smooth muscles (ASMA), F-actin and other autoantibodies.
- Indications: autoimmune hepatitis, primary biliary cirrhosis, rheumatic diseases.
- Initial dilution 1 : 100; polyvalent conjugate anti-human IgAGM, FITC-labelled.
- The BIOCHIP Mosaic™ consists of 6 substrates: human epithelial cells (HEp-2), primate liver, rat kidney, rat liver, rat stomach, VSM47. Thus, a broad spectrum of antigens is present, allowing not only targeted serological diagnoses, but also frequently yielding additional results with clinical relevance.
- Antibodies against cell nuclei (ANA) can be particularly well demonstrated using HEp-2 cells and primate liver, and are identified according to their fluorescence patterns. However, they also stain the cell nuclei of the other tissues more or less intensely. Clinical significance: rheumatic diseases, primary biliary cirrhosis (antibodies against nuclear dots).
- With primate liver, several liver-specific autoantibodies can be investigated e.g. antibodies against liver cell membrane (anti-LMA) and liver-specific protein (anti-LSP). Clinical significance: autoimmune hepatitis.
- Antibodies against mitochondria (AMA) show a granular cytoplasmic fluorescence on all 6 substrates. With the standard substrate rat kidney, the proximal and distal tubule cells fluoresce equally. Clinical significance: primary biliary cirrhosis.
- Autoantibodies against liver-kidney microsomes (anti-LKM) react with rat liver and rat kidney (see below). The other substrates are essentially negative.
- In the case of autoantibodies against smooth muscles (ASMA), the tunica muscularis, the lamina muscularis mucosa as well as the interglandular contractile fibrils fluoresce on the rat stomach. ASMA directed against the target antigen F-actin furthermore react with the cytoskeleton of HEp-2 cells and the bile canaliculi of primate liver. The substrate VSM47 reacts very specifically, showing a filamentous, needle-like fluorescence. Clinical significance: autoimmune (lupoid) chronic active hepatitis.
- The BIOCHIP Mosaic™ can be supplemented as required with additional substrates, e.g. Crithidia luciliae (antibodies against dsDNA), musculus iliopsoas (antibodies against skeletal muscles), heart (antibodies against striated muscles, antibodies against intercalated disks, AMA M7), different EUROPLUS™ substrates (AMA M2, Sp100, SLA/LP, LC-1).
EUROLINE: Profile Autoimmune Liver Diseases
- Incubated EUROLINE Profile Autoimmune Liver Diseases.
- Differentiation of antibodies in autoimmune liver diseases.
- Indications: autoimmune hepatitis, primary biliary cirrhosis, overlap syndromes.
- Serum dilution 1 : 100, conjugate class anti-human IgG, AP-labelled.
- With the EUROLINE Profile Autoimmune Liver Diseases, nine autoantibodies can be determined: antibodies against AMA M2, M2-3E (BPO), Sp100, PML, gp210, LKM-1, LC-1, SLA/LP and Ro-52.
- Test strips can be automatically incubated and evaluated using the systems EUROBlotMaster und EUROLineScan.
| Anti- |
| Associated diseases |
M2, Sp100, PML, gp210 | Primary-biliary cirrhosis | ||
LKM-1, SLA/LP, LC-1 | Autoimmune hepatitis | ||
Ro-52 | Autoimmune hepatitis, rheumatic diseases |
Microplate ELISA: Anti-M2-3E
- Incubated ELISA anti-M2-3E.
- Differentiation of mitochondrial antibodies (AMA).
- Indication: primary biliary cirrhosis (PBC).
- Serum dilution 1 : 100; conjugate class anti-human IgG, POD-labelled.
- Antibodies against M2 can be determined quantitatively in RU/ml.
- Antigen: native pyruvate dehydrogenase complex plus recombinant M2 fusion protein (BPO) containing the immunogenic domains of the E2 subunits of PDH, BCOADH and OGDH.
Microplate ELISA: Anti-SLA/LP, Anti-LC-1, Anti-LKM-1
- Incubated ELISA Anti-SLA/LP, Anti-LC-1, Anti-LKM-1 .
- Monospecific determination of antibodies against soluble liver antigen/liver-pancreas antigen (SLA/LP), cytosolic liver antigen type 1 (LC-1) and liver-kidney microsomes type 1 (LKM-1).
- Indication: autoimmune hepatitis.
- Serum dilution 1 : 100, conjugate class anti-human IgG, POD-labelled.
- 3-point calibration, quantitative.
- Identical incubation conditions and times: all tests can be combined without difficulty on one and the same microplate.
- Recombinant antigens: soluble liver antigen/liver-pancreas antigen (SLA/LP), formiminotransferase-cyclodeaminase (LC-1) and cytochrome P450 IID6 (LKM-1). The corresponding human cDNA was expressed in E. coli (SLA/LP) or insect cells (LC-1, LKM-1).
