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Kit used for quantitative determination anti-nucleosome antibody Ig G via magnetic micro particle chemiluminiscence, and preparation method and detection method thereof

A nucleosome antibody and chemiluminescence technology, applied in the field of immunological detection, can solve the problems of narrow linear range, poor repeatability, and low sensitivity

Inactive Publication Date: 2016-02-03
北京贝尔医疗设备有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Chinese patent CN103105489A discloses an immunoblotting kit for detecting autoimmune disease antibodies and a preparation method thereof, which relates to a immunoblotting kit for detecting antibodies related to various autoimmune diseases, and solves the problem that currently there is no such kit for various autoimmune diseases Technical deficiencies in immunoblotting products for physical examination screening: the nitrocellulose membrane or nylon membrane contains: respectively composed of dsDNA, Sm / RNP, CCP, SSA, SSB, GAD, ICA, IA-2A, TG, AMA-M2 A total of 10 kinds of at least two natural or recombinant antigens coated with more than two parallel detection lines, 1 high-concentration quality control band, 1 medium-concentration quality control band and 1 low-concentration quality control band bring
The patent discloses a membrane strip immunoassay, which can better explain the problems of low sensitivity, narrow linear range, poor repeatability and long reaction time of the membrane strip immunoassay.

Method used

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  • Kit used for quantitative determination anti-nucleosome antibody Ig G via magnetic micro particle chemiluminiscence, and preparation method and detection method thereof
  • Kit used for quantitative determination anti-nucleosome antibody Ig G via magnetic micro particle chemiluminiscence, and preparation method and detection method thereof
  • Kit used for quantitative determination anti-nucleosome antibody Ig G via magnetic micro particle chemiluminiscence, and preparation method and detection method thereof

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Effect test

Embodiment 1

[0063] A kit for quantitatively detecting anti-nucleosome antibody IgG using magnetic particle chemiluminescence, the kit comprising: anti-nucleosome antibody IgG calibrator, anti-nucleosome antibody IgG reagent No. 1, anti-nucleosome antibody IgG reagent No. 2, anti-nucleosome antibody IgG magnetic separation reagent, anti-nucleosome antibody IgG quality control substance and cleaning solution.

[0064] Analytical performance evaluation of kit of the present invention:

[0065] 1) Accuracy evaluation

[0066] Add anti-nucleosomal antibody IgG sample A with a concentration of about 200RU / mL, which allows a concentration deviation of ±20%, to serum or other corresponding matrix sample B, and the volume of A added does not exceed the total volume of A+B 10%, calculate the recovery rate R according to formula (1), the recovery rate of this method is in the range of 85-115%, see Table 1 for the data.

[0067] R = C ×...

Embodiment 2

[0109] A preparation method of a kit for quantitative detection of anti-nucleosome antibody IgG using magnetic particle chemiluminescence comprises the following steps:

[0110] Step 1. The preparation of the anti-nucleosome antibody IgG calibrator comprises: the anti-nucleosome antibody IgG calibrator dilution preparation step and the anti-nucleosome antibody IgG calibrator preparation step:

[0111] The preparation step of described anti-nucleosome antibody IgG calibrator dilution comprises:

[0112] Add 800mL of purified water and 11.2g of Tris to the container, stir and mix evenly, add 8.5g of sodium chloride and 2mL of Proclin300, stir until completely dissolved, adjust the pH value to 7.0-7.5, add 40g of bovine serum white Put the protein into the container, stir until completely dissolved, then adjust the pH value to 7.0-7.5, dilute to 1L with purified water, filter with a 0.2μm filter, and store the obtained dilution of the calibration product at 2-8°C until use;

[...

Embodiment 3

[0131] On the basis of Example 2, the kit also includes a cleaning solution, the preparation steps of the cleaning solution:

[0132]Add 800mL of purified water, Tris12.1g and NaCl8.5g to a 1L container, stir until completely dissolved; then add 5g of Tween-20 and 5g of Triton100, stir until completely mixed, and adjust the pH value to 7.5 -8.0, constant volume 1L, filtered with a 0.2μm filter, and stored at a temperature range of 2-8°C.

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Abstract

The invention belongs to the technical field of immunological detection, and more specifically relates to a kit used for quantitative determination anti-nucleosome antibody Ig G via magnetic micro particle chemiluminiscence, and a preparation method and a detection method thereof. The kit comprises an anti-nucleosome antibody Ig G calibration material, an anti-nucleosome antibody Ig G reagent 1, an anti-nucleosome antibody Ig G reagent 2, an anti-nucleosome antibody Ig G magnetic separation reagent, an anti-nucleosome antibody Ig G quality control product, and a cleaning solution. The invention also discloses the preparation method and the detection method of the kit. The detection method is invented based on conventional membrane strip immunization and enzyme linked immunosorbent assay, sensitivity and linearity range are increased 10<3> to 10<5>, quantitative determination is realized, sensitivity, specificity, and accuracy are high, cost is low, operation is simple and convenient, result judgment is objective, fully automatic application is realized via combination with an automatic chemicaluminescence immunity analyzer, and application prospect is promising.

Description

technical field [0001] The invention relates to the technical field of immunological detection, in particular to a kit for quantitatively detecting anti-nucleosome antibody IgG by using magnetic particle chemiluminescence, and a preparation method and a detection method thereof. Background technique [0002] Anti-nucleosome antibody (anti-nucleosomeantibody, AnuA) is mainly an autoantibody against nucleosome molecules produced by B cells. Nucleosome is the functional subunit of chromosome, which is a complex formed by DNA and histone, and exists in the nucleus. In systemic lupus erythematosus (SLE), nucleosome is the main autoantigen, and anti-nucleosome antibody is one of the early indicators of SLE. Many literatures believe that anti-nucleosome antibody is a good marker for the diagnosis of SLE, which is superior to anti-dsDNA, with high sensitivity and high specificity. Studies have shown that nucleosomes are the main autoantigens of SLE, which may be directly related t...

Claims

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Application Information

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IPC IPC(8): G01N33/543
CPCG01N33/54326
Inventor 不公告发明人
Owner 北京贝尔医疗设备有限公司
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