A kit for detecting anti-mutant citrullinated vimentin antibody

A technology of citrullination and vimentin, applied in the field of medical immunity, can solve the problems of low sensitivity and specificity, complicated detection steps, and long reaction time of the colloidal gold method, and achieve small coefficient of variation, improved specificity, and good specificity Effect

Active Publication Date: 2022-03-01
AUTOBIO DIAGNOSTICS CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The ELISA method requires special equipment, the reaction time is relatively long, and the detection steps are complicated; the colloidal gold method has low sensitivity and specificity, and its application is limited

Method used

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  • A kit for detecting anti-mutant citrullinated vimentin antibody
  • A kit for detecting anti-mutant citrullinated vimentin antibody
  • A kit for detecting anti-mutant citrullinated vimentin antibody

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Effect test

Embodiment 1

[0038] Embodiment 1 The present invention detects the kit of anti-mutant citrullinated vimentin antibody

[0039] 1. Preparation of magnetic particles coupled with MCV antigen

[0040] Take 30ul of magnetic bead buffer (0.01M, pH 6.0 MES buffer), blow and beat magnetic beads (particle size about 1um) repeatedly, place on a magnetic bead separator, discard the supernatant, add carbodiimide activation solution, React at room temperature for 1 hour; then add MCV antigen for coupling overnight; after coupling, discard the supernatant on the magnetic bead separator, add ethanolamine solution to block for 30 minutes to 1 hour; place the magnetic beads on the separator, discard the upper After cleansing, first add blocking solution 1 (0.01M Tris-Nacl buffer at pH 8.0 containing 2% casein and 0.5% Tetronic 1307) to mix evenly, place in a magnetic bead separator, discard the supernatant, repeat 3-5 times Afterwards, use blocking solution 2 to store at 2-8°C. Wherein, the blocking sol...

Embodiment 2

[0049] The detection method of embodiment 2 kit of the present invention

[0050] 1. Take a 20ul sample and dilute it with sample diluent 1:9;

[0051] 2. Take 50ul diluted sample, add 20ul magnetic particle complex reagent, pipette and mix well, then add 100ul enzyme conjugate solution, incubate at 37°C for 15min;

[0052] 3. Add 0.02M PBS washing solution to the above mixed solution, wash 5 times, add substrate (50ul each of solution A and solution B), and then detect in Antu automatic immunochemiluminescence instrument A2000.

Embodiment 3

[0053] The performance detection of embodiment 3 kits of the present invention

[0054] 1. Linear range:

[0055] Control kit 1: The magnetic particles coupled with MCV antigen are blocked by one-step method. The specific method is: take 30ul of magnetic bead buffer, blow and beat the magnetic beads (particle size is about 1um) repeatedly, place them on the magnetic bead separator, discard After supernatant, add carbodiimide activation solution and react at room temperature for 1 hour; then add MCV antigen for overnight coupling; after coupling, discard the supernatant on the magnetic bead separator, add ethanolamine solution for blocking for 30 minutes to 1 hour; The magnetic beads are also placed on the separator. After discarding the supernatant, first add blocking solution 1 (0.01M Tris-Nacl buffer solution at pH 8.0 containing 2% casein and 0.5% Tetronic 1307) to mix evenly, and place on the magnetic beads. Separator, discard the supernatant, repeat 3-5 times, use blocki...

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Abstract

The invention relates to the field of medical immunity, in particular to a kit for detecting anti-mutant citrullinated vimentin antibody. The kit includes: sample diluent, magnetic particles coupled with MCV antigen, enzyme conjugate and luminescent substrate; the enzyme conjugate is composed of enzyme-labeled mouse anti-human IgG antibody and enzyme conjugate diluent; the Sample diluent is 0.05M at pH 7.4 containing 0.5%‑2% bovine serum albumin, 0.5%‑1% Casein protein, 100‑500ng / ml heterophile antibody blocker and 0.1%‑5% metal chelator Phosphate buffer. The kit of the invention has high sensitivity, good specificity, wide linear range, good stability and convenient use. In addition, this kit can be equipped with an automated system, which is easy to operate and has a large throughput; it adopts a magnetic particle platform, which has a wide range of applications and low cost.

Description

technical field [0001] The invention relates to the technical field of medical immunity, in particular to a kit for detecting mutant citrullinated vimentin antibodies. Background technique [0002] Rheumatoid arthritis (RA) is an autoimmune disease mainly characterized by chronic inflammation of the joints, with a high disability rate. The deposition of citrulline-related autoantibodies in the joint cavity is the main reason for the persistence of RA synovitis. In addition, genetic factors and environmental factors are also included. The diagnosis of RA still adopts the RA diagnostic classification criteria revised by the American Rheumatology Association (ARA) (1987), in which rheumatoid factor (RF) is a relatively common serological indicator, and the IgM type of RF is specific in the diagnosis of RA. It is not high, and it can also be detected in other autoimmune diseases, infections and some elderly people. At present, anti-citrulline synthetic protein (ACPAS) has beco...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/535G01N33/543G01N21/76G01N33/68
CPCG01N33/535G01N33/54326G01N21/76G01N33/68
Inventor 李奎于林李双法丁蒙蒙刘珂毛宁宇付光宇
Owner AUTOBIO DIAGNOSTICS CO LTD
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