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Anti-double-stranded DNA antibodies IgG chemiluminescence immunoassay kit and preparation method thereof

A chemiluminescence and reagent kit technology, which is applied in the direction of measuring devices, scientific instruments, instruments, etc., can solve the problems of narrow detection linear range, low detection sensitivity, and high detection cost, and achieve high sensitivity, wide linear range, and easy operation.

Inactive Publication Date: 2017-02-22
SHENZHEN YHLO BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0013] The current anti-double-stranded DNA antibody IgG detection technology has the following disadvantages: high detection cost, low detection sensitivity, narrow detection linear range, low reproducibility, inability to quantify, complicated operation, etc.

Method used

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  • Anti-double-stranded DNA antibodies IgG chemiluminescence immunoassay kit and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Example 1: Preparation method of anti-double-stranded DNA antibody IgG chemiluminescence immunoassay kit

[0024] (1) Preparation of nano-magnetic beads coated with anti-double-stranded DNA antibody IgG monoclonal antibody:

[0025] Take 50mg of carboxylated magnetic particles (particle size: 0.05-1um) suspension, magnetically separate to remove the supernatant, resuspend with 0.02 M, pH 5.5MES buffer, add 0.5-2mL of newly prepared 10 mg / mL EDC Aqueous solution to activate the carboxyl groups on the surface of magnetic beads, add 3-5mg anti-double-stranded DNA antibody IgG monoclonal antibody, suspend at room temperature for 2-10 h, magnetically separate, remove the supernatant, and use 0.1 M pH 8.0 containing 2% BSA Tris buffer was resuspended to 1 mg / mL to obtain magnetic particles coated with anti-double-stranded DNA antibody IgG monoclonal antibody, and each bottle was divided into 5 mL bottles and stored at 4°C for later use.

[0026] (2) Preparation of acridinium...

Embodiment 2

[0034] Embodiment 2: Anti-double-stranded DNA antibody IgG chemiluminescent immunoassay method:

[0035] The present invention uses a fully automatic chemiluminescence immunoassay analyzer as a detection tool, and the methodological mode of the present invention is a competitive method, that is, the instrument sequentially adds 20 uL of samples and 50 uL of magnetic particles coated with anti-double-stranded DNA antibody IgG monoclonal antibody And 50uL of acridinium ester coated with anti-double-stranded DNA antibody IgG. After reacting for 10 minutes, perform magnetic separation. The instrument sends the reaction mixture into the darkroom, and sequentially adds 50uL of chemiluminescence pre-excitation solution and 50uL of chemiluminescence excitation solution for luminescence reaction. , and finally record the luminous intensity, and calculate the anti-double-stranded DNA antibody IgG content of the tested sample from the standard curve.

Embodiment 3

[0036] Example 3: Performance evaluation of anti-double-stranded DNA antibody IgG chemiluminescent immunoassay kit

[0037] The detection curve is attached figure 1 .

[0038] Sensitivity detection:

[0039] The sensitivity of the anti-double-stranded DNA antibody IgG chemiluminescence immunochromatography kit was calculated according to the recommended experimental protocol in the CLSI EP17-A document, and the obtained sensitivity was 2.0 IU / mL.

[0040] Linear detection:

[0041] Perform linear analysis on standard substances with concentrations of 0 IU / mL, 20 IU / mL, 40 IU / mL, 80 IU / mL, 160 IU / mL, and 320 IU / mL, and calculate the linear correlation coefficient, r=0.9985. In addition, the The linear range of the kit for detection of anti-double-stranded DNA antibody IgG samples is 2-300 IU / mL.

[0042] Precision determination:

[0043]Take two anti-double-stranded DNA antibody IgG samples with a concentration of 50 IU / mL and 200IU / mL, make three parallels for each concen...

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Abstract

The invention discloses an anti-double-stranded DNA antibodies IgG chemiluminescence immunoassay kit. The kit comprises biotinylated double-stranded DNA antigens, streptavidin-coated magnetic particles, acridine ester-labeled murine anti-human IgG monoclonal antibodies, anti-double-stranded DNA antibody IgG scaffolds, pre-excitation fluid and excitation fluid. Moreover a preparation method of the anti-double-stranded DNA antibodies IgG chemiluminescence immunoassay kit is also disclosed. Compared with the current kit the kit has the advantages of simple operation, high sensitivity and wide detection scope and the like.

Description

technical field [0001] The invention relates to the field of in vitro diagnostic immunodetection. Specifically, the invention provides a chemiluminescence immunodetection anti-double-stranded DNA antibody IgG kit and a preparation method thereof. Background technique [0002] Double-stranded deoxyribonucleic acid (double stranded DNA, dsDNA) antibody is an autoantibody that can bind to natural DNA. Anti-double-stranded DNA antibodies exist in the serum of more than 90% of active systemic lupus erythematosus (SLE) patients, and double-stranded DNA antibodies are closely related to the pathological changes, activity, curative effect and prognosis of SLE. [0003] SLE is an autoimmune disease that seriously endangers human health. It often occurs in young women and can affect various systems of the body. The important immunological feature of SLE disease is the production of autoantibodies, which are not only of great value in the diagnosis of the disease, but also play an imp...

Claims

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Application Information

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IPC IPC(8): G01N33/543G01N33/577
CPCG01N33/54326G01N33/577
Inventor 郝建华徐向红夏福臻钱纯亘代洪飞
Owner SHENZHEN YHLO BIOTECH
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