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Marker and its application, kit, detection method of the marker

A kit and marker technology, applied in the biological field, can solve the problem of low specificity of RF detection and achieve high sensitivity, short time-consuming and simple method

Active Publication Date: 2021-02-05
GUANGZHOU LDEBIO TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] At present, the detection specificity of RF is low; the detection of ACPA using artificially synthesized citrullinated peptides can only reflect the level of self-generated antibodies, and the level of ACPA may not be associated with the disease state

Method used

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  • Marker and its application, kit, detection method of the marker
  • Marker and its application, kit, detection method of the marker
  • Marker and its application, kit, detection method of the marker

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0098] Example 1 Determination of the level of citrullinated antigen-antibody immune complex (ECP) at different dilutions in a case of RA patient serum

[0099] 1) Serum preparation

[0100] Fresh blood from RA patients was collected in blood collection tubes without any anticoagulant, centrifuged at 3000 rpm for 10 min, and the supernatant was collected as serum, which was stored at -20°C to -80°C for later use.

[0101] 2) ECP Elisa detection kit preparation

[0102] Coated microplate; sample diluent (5% skimmed milk powder); washing solution (0.5% Triton X-100); enzyme-labeled antibody; chromogenic solution (TMB); stop solution (1M sulfuric acid).

[0103] 3) Experimental operation steps

[0104] A. Dilute the serum sample 10 times, 20 times, 50 times, 100 times with the sample diluent; add it to the sample well of the reaction plate respectively, and the volume of each well is 100ul;

[0105] B. Incubate at room temperature (25°C) for 1 hour;

[0106] C. Add 200ul of w...

Embodiment 2

[0120] Example 2 Comparison of citrullinated antigen-antibody immune complex (ECP) levels in the serum of 35 normal people and 45 RA patients

[0121] 1) Serum preparation

[0122] Fresh blood from normal subjects and RA patients was collected in blood collection tubes without any anticoagulant, centrifuged at 3000 rpm for 10 min, and the supernatant was collected as serum, which was stored at -20°C to -80°C for later use.

[0123] 2) ECP Elisa detection kit preparation

[0124] Coated microplate; sample diluent (5% skimmed milk powder); washing solution (0.5% Triton X-100); enzyme-labeled antibody; chromogenic solution (TMB); stop solution (1M sulfuric acid).

[0125] 3) Experimental operation steps

[0126] A. Dilute the serum sample 10 times with the sample diluent; add it to the sample wells of the reaction plate respectively, and the volume of each well is 100ul; select the remaining wells and add 100ul each of the negative reference sample and the positive reference sa...

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Abstract

The invention relates to the field of biotechnology, in particular to a marker and its application, a kit, and a detection method for the marker. The present invention uses the principle that endogenous citrullinated antigens and ACPA can form antigen-antibody immune complexes to search for and explore a detection method and related ELISA kits for citrullinated antigen-antibody immune complexes, which can not only reflect the level of ACPA, It can also reflect the level of the citrullinated antigen-antibody immune complex, and the citrullinated antigen-antibody immune complex is associated with the state of the disease, and the change in its amount can directly reflect the progress of the disease and the evaluation of the treatment effect, so as to achieve rapid, The purpose of accurate and convenient diagnosis and evaluation of RA.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a marker and its application, a kit, and a detection method for the marker. Background technique [0002] Rheumatoid Arthritis (RA) is a chronic autoimmune disease whose main pathological changes are synovial inflammation. The pathological process of RA is closely related to the production of a variety of autoantibodies in patients and the formation of antigen-antibody immune complexes (ICs) with corresponding antigens. Some ICs in the circulation can be repeatedly deposited in the synovial tissue of the joints, which can be caused by activating the cascade reaction of the complement system. Inflammation of the synovium, causing joint damage. ICs in the synovial fluid and serum of RA patients can promote the growth of chondrocytes, the production of NO and apoptosis, suggesting that ICs play an important role in the pathogenesis of RA. ICs have high sensitivity in the detection of ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N21/31G01N33/68
CPCG01N21/31G01N33/68G01N2800/52
Inventor 楼建荣
Owner GUANGZHOU LDEBIO TECH CO LTD
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