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Specific antigenic mark for rheumatoid arthritis and its use

A technology for rheumatoid arthritis and arthritis, applied in rheumatoid arthritis-specific antigenic markers and their application fields, can solve the problems of unclear pathological mechanism and untargeted treatment, achieve good practical value, and be easy to popularize and use , the effect of sensitive detection methods

Inactive Publication Date: 2005-12-28
SHANDONG MEDICAL BIO TECH RES CENT
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Like other autoimmune diseases, there is still no very effective targeted treatment, and its pathological mechanism is still unclear

Method used

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  • Specific antigenic mark for rheumatoid arthritis and its use
  • Specific antigenic mark for rheumatoid arthritis and its use
  • Specific antigenic mark for rheumatoid arthritis and its use

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Example 1: Determination of citrullinated fibronectin in patient blood

[0028] (1) Plasma preparation.

[0029] Fresh blood from rheumatoid arthritis patients, systemic lupus erythematosus patients and normal subjects was collected with blood collection tubes containing sodium citrate at a final concentration of 3.8%. Centrifuge at 1000X g for 30 minutes. Collect the supernatant. Plasma can be stored at -80°C.

[0030] (2) Solution preparation

[0031] PBS buffer formula: 8g / L NaCl, 0.2g / L KCl, 1.15g / L NaHPO 4 and 0.2g / L KH2PO 4 , pH7.5;

[0032] PBS-Tween eluent: prepare by adding Tween 20 into PBS buffer at a ratio of 1:1000, and mix well.

[0033] (3) Steps:

[0034] A. Dilute the anti-fibrin monoclonal antibody (Abcam product) 4000 times with 0.05M, pH9.6 carbonic acid / bicarbonate (carbonate-bicarbonate) buffer solution, and drop 100ul antibody solution in the sample well of the 96-well ELISA reaction plate , place the plate at 4°C for 12 hours to cross-link...

Embodiment 2

[0044] Example 2: Determination of citrullinated fibronectin in patient blood

[0045] (1) Plasma preparation.

[0046] Fresh blood from rheumatoid arthritis patients, systemic lupus erythematosus patients and normal subjects was collected with blood collection tubes containing sodium citrate at a final concentration of 3.8%. Centrifuge at 8000Xg for 30 minutes. Collect the supernatant. Plasma can be stored at -80°C.

[0047] (2) Solution preparation

[0048] PBS buffer formula: 8g / L NaCl, 0.2g / L KCl, 1.15g / L NaHPO 4 and 0.2g / L KH2PO 4 , pH7.6;

[0049] PBS-Tween eluent: prepare by adding Tween 20 into PBS buffer at a ratio of 1:1000, and mix well.

[0050] (3) Steps:

[0051] A. Dilute anti-fibrin monoclonal antibody (Abcam company product) 3000 times with 0.05M, pH9.6 carbonic acid / bicarbonate (carbonate-bicarbonate) buffer solution, drop 100ul antibody solution in the sample hole of 96-well ELISA reaction plate , place the plate at 4°C for 10-12 hours to cross-link...

Embodiment 3

[0061] Embodiment 3: kit composition

[0062] 1. Anti-fibronectin pre-coated ELISA plate (strip)

[0063] Take the anti-fibronectin antibody at a ratio of 1:3000, dilute it with pH9.6 carbonate buffer, add 100ul to each well, let it stand at room temperature for 4 hours, wash 3 times with PBS containing 0.05% Tween-20, and add 100ul to each well. Add 200 ul of 2% BSA and place at 37°C for 2 hours. After buckling dry, blow dry, vacuum seal in a tin-platinum bag, and store at 4°C.

[0064] 2. Anti-citrulline antibody, routinely labeled with horseradish peroxidase.

[0065] 3. Product diluent: containing 2% BSA, PBS.

[0066] 4. Washing liquid: containing 0.05% Tween-20PBS.

[0067] 5. Color developer: 0.4% TMB.

[0068] 6. Stop solution: 2% H 2 SO 4 .

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Abstract

A mark substance of specificity - anti-genicity for atrophic arthritis is carbamyl ornithine fibre connexin existied in blood plasma and joint synovium tissue of atrophic arthritis patient. The invented mark substance has excellent application value in preparing reagent and medicine for diagnosing and curing disease of atrophic arthritis.

Description

technical field [0001] The present invention relates to a rheumatoid arthritis marker and its application, in particular to a rheumatoid arthritis specific antigenic marker and its application. technical background [0002] Rheumatoid arthritis is a kind of chronic polyarthritis, and the incidence rate of Chinese people is 5 / 1000. The disease manifests as joint swelling and pain, and in the advanced stage, the joint tissue is severely damaged and the joint function is lost. It is reported that about 4 million new cases of rheumatoid arthritis are added every year in China. Like other autoimmune diseases, there is still no very effective targeted treatment, and its pathological mechanism is still unclear. Rheumatoid arthritis is an autoimmune disease. So far, clinically, various autoimmune antibodies (rheumatoid factor, anti-keratin filament protein antibody, anti-keratin antibody, anti-perinuclear factor and anti-cyclic citrulline antibody) in the patient's blood are dete...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/52
Inventor 常晓天朱有名韩金祥
Owner SHANDONG MEDICAL BIO TECH RES CENT
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