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Composite detection antigen and application thereof

A technology for compound detection and combination of antigens, applied to detection kits and their application in the detection of rheumatoid arthritis And other issues

Inactive Publication Date: 2018-01-26
GUANGZHOU LDEBIO TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0008] The object of the present invention is to provide a composite detection antigen and its application. The detection antigen is a combination of citrullinated protein and carbamylated protein, which can detect multiple RA autoantibodies at one time, and overcomes the sensitivity and limitations of single autoantibody detection. Insufficient specificity and the cumbersome operation of a single detection of multiple autoantibodies one by one greatly improve the detection efficiency and the accuracy of result judgment

Method used

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  • Composite detection antigen and application thereof
  • Composite detection antigen and application thereof
  • Composite detection antigen and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0067] Embodiment 1: the assembly of kit

[0068] 1. Main reagents:

[0069] (1) Coating buffer: 0.01M-0.5M pH9.0-pH9.6 carbonate buffer, 0.01%-0.5% ProClin300, pH 6.9-9.4;

[0070] (2) Blocking solution: 0.5-10% BSA, 0.01M-0.5M PBS buffer, 150mM NaCl, 0.01%-0.5% ProClin300, pH 6.9-9.4;

[0071] (3) Sample diluent: 0.5-10% BSA, 0.01M-0.5M Tri-HCL buffer, 0.01%-0.5% ProClin300, pH 6.9-9.4;

[0072] (4) Diluent of negative and positive reference substances: 0.5-10% BSA, 0.01M-0.5M PBS buffer, 0.01%-0.5% ProClin300, pH 6.9-9.4;

[0073] (5) Washing solution: 0.01%-0.2% Tween-20, 0.01M-0.5M PBS buffer, 0.01%-0.5% ProClin300, pH 6.9-9.4;

[0074] (6) Enzyme-labeled antibody diluent: 0.5-10% BSA, 0.01M-0.5M PBS buffer, 0.01%-0.5% ProClin300, pH 6.9-9.4;

[0075] (7) Chromogenic buffer: 0.01M-1M phosphate-citrate buffer, pH 3.0-5.0;

[0076] (8) Chromogenic solution: TMB;

[0077] (9) Termination solution: sulfuric acid.

[0078] 2. Preparation of reaction plate working soluti...

Embodiment 2

[0085] Embodiment 2: the assembly of kit

[0086] 1. Main reagents:

[0087] (1) Coating buffer: 0.01M-0.5M PBS buffer, 0.01%-0.5% ProClin300, pH6.9-9.4;

[0088] (2) Blocking solution: 0.5-10% BSA, 0.01M-0.5M PBS buffer, 0.01%-0.5% ProClin300, pH6.9-9.4;

[0089] (3) Sample diluent: 0.5-10% BSA, 0.01M-1M Tri-HCL buffer, 0.01%-0.5% ProClin300, pH 6.9-9.4;

[0090] (4) Diluent of negative and positive reference substances: 0.5-10% BSA, 0.01M-0.5M PBS buffer, 0.01%-0.5% ProClin300, pH 6.9-9.4;

[0091] (5) Washing solution: 0.01%-0.2% Tween-20, 0.01M-0.5M PBS buffer, 0.01%-0.5% ProClin300, pH 6.9-9.4;

[0092](6) Enzyme-labeled antibody diluent: 0.5-10% BSA, 0.01M-0.5M PBS buffer, 0.01%-0.5% ProClin300, pH 6.9-9.4;

[0093] (7) Chromogenic buffer: 0.01M-1M phosphate-citrate buffer, pH 3.0-5.0;

[0094] (8) Chromogenic solution: TMB;

[0095] (9) Termination solution: sulfuric acid.

[0096] 2. Preparation of reaction plate working solution

[0097] Two types of post-tran...

Embodiment 3

[0103] Embodiment 3: the usage method of kit

[0104] (1) Pretreatment: place each component of the kit at room temperature to equilibrate for 0.5-1h, and dilute the sample 1:50 with the sample diluent;

[0105] (2) Adding samples: Add 100 μl / well of the diluted sample to be tested and negative and positive controls to the corresponding wells of the microplate, and seal the plate with a sealing film;

[0106] (3) Incubation: place the loaded microtiter plate on a shaker at 200rpm-400rpm and incubate at room temperature for 30-60min;

[0107] (4) Washing: the incubated ELISA plate was discarded and patted dry, and washed 4-6 times with washing liquid;

[0108] (5) Add enzyme-labeled antibody: add enzyme-labeled antibody to the enzyme-labeled plate at 100 μl / well, and seal the plate with a sealing film;

[0109] (6) Incubation: Place the added ELISA plate on a 200rpm-400rpm shaker and incubate at room temperature for 30-60min;

[0110] (7) Washing: the incubated ELISA plate w...

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Abstract

The invention relates to a composite detection antigen and application thereof, in particular to a composite rheumatoid arthritis detection antigen and application thereof. The detection antigen is acombined antigen of citrullinated protein and carbamylated protein, a kit composed of the detection antigen can detect multiple rheumatoid arthritis autoantibodies at a time, overcomes the disadvantages that single autoantibody detection has inadequate sensitivity and specificity and one-by-one detection of multiple autoantibodies is tedious in operation, and greatly improves the detection efficiency and result judgment accuracy, moreover, the kit provided by the invention also can be used for clinical examination of rheumatoid arthritis and post-treatment curative effect assessment, thus having broad market prospects and huge economic benefits.

Description

technical field [0001] The invention relates to the technical field of biological immune detection, in particular to a composite detection antigen and its application, in particular to a composite detection antigen, a detection kit and its application in the detection of rheumatoid arthritis. Background technique [0002] Rheumatoid arthritis (RA) is a common and difficult-to-cure disease that causes irreversible damage to the joints. The late stage of the disease often leads to disability, affects the quality of life, and loses the ability to work. It is characterized by persistent synovitis, systemic inflammation, and the presence of autoantibodies. At present, the etiology and pathogenesis of RA are still poorly understood, and timely diagnosis and early treatment are of great significance for the control of the disease. At present, there are many clinical methods for diagnosing RA. The 2010 American College of Rheumatology (ACR) diagnostic criteria stipulates the number...

Claims

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Application Information

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IPC IPC(8): G01N33/68
Inventor 黄小燕杨翔李杨茹志伟楼建荣周单
Owner GUANGZHOU LDEBIO TECH CO LTD
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