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Background
The diagnostic feature of any autoimmune disease is the presence of autoantibodies against a particular or several self-antigens. Waaler has developed an idea of autoimmunity in rheumatoid arthritis (RA). He found that any irregularities in connective tissue contribute to the disease.
Autoantibody rheumatoid factor (RF) is increased in RA patients. Steffen also found that RA might be a collagen autoimmune disorder in 1970. Based on the clinical studies, in reaction to the RA etiology, many autoantibodies are produced like anti-collagen antibodies and RF in the synovial plasma cells. RF is still commonly used in the diagnostic treatment for RA.
The further research led to new markers like anti-keratin antibody, anti-RA33 and anti-citrullinated antibodies. They showed a specificity in the test of immunodiagnostic RA. Anti-keratin and anti-citrullinated protein antibodies are present in the perinuclear area of the cell.
Anti-RA33 antibody is present in the nucleolar area of the cell. Cordonnier researcher has tried to connect the RF, anti-RA33, and anti-CCP in the early stage of RA.
Positive RF, anti-CCP, and AKA were reduced in the early stage of RF. This was not occurred for the anti-RA33 antibody. This led to the important discovery that anti-RA33 antibody is the promising marker to diagnose the RA in the early stage.
Hassfeld discovered the anti-RA33 antibody in 1989. It was discovered to bind a specific nuclear protein in the HeLa cells. Anti-RA33 antibody targets a spliceosome the hn-RNP A2 protein in mice models who has RA. This protein with variation of B1 and B2 acts as an autoantigen in RA patients. Many epitopes of hn-RNP are linked to the sequence of protein which associated with the premature mRNA in hn-RNP complex.
Other hn-RNP family also involved in the autoantigens in autoimmune rheumatoid disease. Hn-RNP A2 is found in the lymph nodes, brain, skin, and reproductive organs. It absents in healthy joint synovium.
Anti-RA33 antibody shows specificity from 69 to 96 %. Anti-RA33 antibody shows sensitivity from 6 to 58%. It could be because of the severity of RA, ethnic background, or the quality of purification of anti-RA33 used as an autoantigen in ELISA test.
Indications/Applications
The diagnostic feature of any autoimmune disease is the presence of autoantibodies against a particular or several self-antigens. Waaler has developed an idea of autoimmunity in rheumatoid arthritis (RA). He found that any irregularities in connective tissue contribute to the disease.
Autoantibody rheumatoid factor (RF) is increased in RA patients. Steffen also found that RA might be a collagen autoimmune disorder in 1970. Based on the clinical studies, in reaction to the RA etiology, many autoantibodies are produced like anti-collagen antibodies and RF in the synovial plasma cells. RF is still commonly used in the diagnostic treatment for RA.
The further research led to new markers like anti-keratin antibody, anti-RA33 and anti-citrullinated antibodies. They showed a specificity in the test of immunodiagnostic RA. Anti-keratin and anti-citrullinated protein antibodies are present in the perinuclear area of the cell.
Anti-RA33 antibody is present in the nucleolar area of the cell. Cordonnier researcher has tried to connect the RF, anti-RA33, and anti-CCP in the early stage of RA.
Positive RF, anti-CCP, and AKA were reduced in the early stage of RF. This was not occurred for the anti-RA33 antibody. This led to the important discovery that anti-RA33 antibody is the promising marker to diagnose the RA in the early stage.
Hassfeld discovered the anti-RA33 antibody in 1989. It was discovered to bind a specific nuclear protein in the HeLa cells. Anti-RA33 antibody targets a spliceosome the hn-RNP A2 protein in mice models who has RA. This protein with variation of B1 and B2 acts as an autoantigen in RA patients. Many epitopes of hn-RNP are linked to the sequence of protein which associated with the premature mRNA in hn-RNP complex.
Other hn-RNP family also involved in the autoantigens in autoimmune rheumatoid disease. Hn-RNP A2 is found in the lymph nodes, brain, skin, and reproductive organs. It absents in healthy joint synovium.
Anti-RA33 antibody shows specificity from 69 to 96 %. Anti-RA33 antibody shows sensitivity from 6 to 58%. It could be because of the severity of RA, ethnic background, or the quality of purification of anti-RA33 used as an autoantigen in ELISA test.
Reference Range
The result will be positive for anti-RA33 antibodies if it the value is 25 U/mL or above.
Interpretation
Positive test result indicates the presence of RA. The level is falsely increased in patients with autoimmune disorders like mixed connective tissue disease, juvenile idiopathic arthritis, and systemic sclerosis.
Anti-RA33 antibody found in the early stage of RA. It is associated with the disease severity. The limiting factor is the sensitivity range. The values are 6% to 75%.
Sieghart researched that IgG-RA33, IgM- RA33, and IgA-RA33 antibodies specificity in RA is 97.2%, 95.8%, and 97.5%.
Collection And Panels
Sample type: Human plasma, serum, or cell culture supernatant. Organizations who have recombinant and natural hn-RNP/RA33 concentrations.
Sample collection method:
Serum: Serum separator tube.
Allow the blood samples to clot for at least 2 hours at the normal temperature or at 4°C for overnight. Centrifuge the samples at 1000×g for 15 minutes. Separate the serum and perform the assay.
Plasma: EDTA or heparin anticoagulant-containing tube.
Centrifuge the blood samples with the 30 minutes at 1000×g at 2 to 8 °C. Separate the plasma and perform the assay.
Tissue Homogenates: Rinse homogenize in 1 mL of 1×PBS, 100 mg tissue with 1×PBS. Store it at -20 °C for overnight. Perform 2 freeze and thaw cycles to breakdown the cell membranes. Perform centrifugation process to homogenates at 5000×g for 5 min. at 2 °C to 8 °C. Separate the supernatant and perform the assay.
Sample volume: 50 to 100 ÎĽL
Sample storage: Samples must be used with in the 5 days of collection. It is stored at 2 °C to 8 °C or at -20 °C to -80 °C to protect the samples from the contamination and loss of bioactivity.
Rejected samples: Hemolyze samples are not acceptable.
Method used: ELISA method (enzyme-linked immunosorbent assay) with double sandwich ELISA
Considerations: The level of the detection is 10 to 0.156 ng/mL. The level of sensitivity of detectable hn-RNP/RA33 is up to 0.05 ng/mL. The accuracy of this test is 8% or less than that. There is limited knowledge and skills. It is not possible to do the cross reactivity between the target antigen and analogues of the other organisms.
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