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Magnetic particle-based quantitative chemiluminescent assay kit for anti-histone antibody IgG, and preparation and detection methods thereof

A chemiluminescence and histone technology, which is applied in chemiluminescence/bioluminescence, analysis by making materials react chemically, measurement devices, etc., can solve problems such as low sensitivity, no detection, narrow linear range, etc.

Inactive Publication Date: 2016-09-21
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

Membrane immunoassay uses membrane strip chromogenic technology, which is characterized by fixing several items in the same membrane strip for measurement, generally through manual or semi-automatic membrane strip instrument for experimental operation, and finally qualitative judgment by naked eyes, this technology has low sensitivity, The reaction time is long, the detection items can only be fixed and combined, and the flexibility is poor
The sensitivity of enzyme-linked immunosorbent assay has been improved on the basis of membrane strip immunoassay, but it is still low, and the linear range is narrow, the repeatability is poor, and the reaction time is long, which still cannot meet the clinical application well.
[0006] At present, the application of magnetic particle chemiluminescence analysis in anti-histone antibody IgG immunoassay products has not yet been seen.

Method used

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  • Magnetic particle-based quantitative chemiluminescent assay kit for anti-histone antibody IgG, and preparation and detection methods thereof
  • Magnetic particle-based quantitative chemiluminescent assay kit for anti-histone antibody IgG, and preparation and detection methods thereof
  • Magnetic particle-based quantitative chemiluminescent assay kit for anti-histone antibody IgG, and preparation and detection methods thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0069] Preparation of anti-histone antibody IgG calibrator:

[0070] Step 1) Prepare anti-histone antibody IgG calibrator dilution:

[0071] Add 800ml of purified water, 11.2g of Tris, 8.6g of sodium chloride and 2ml of Proclin300 into the container, stir well until completely dissolved; use 4M HCL to adjust the pH of the solution to 7.0-7.5; add 40g of bovine serum albumin Add to the container, stir well until completely dissolved; then adjust the pH of the solution to 7.0-7.5 with 4M HCL; dilute the solution to 1L with purified water, and filter with a 0.2μm filter to obtain the anti-histone antibody IgG calibrator dilution solution, stored at 2-8°C for later use;

[0072] Step 2) Prepare anti-histone antibody IgG calibrator:

[0073] Dilute anti-histone antibody IgG with anti-histone antibody IgG calibrator diluent to each concentration point of 0, 5, 20, 50, 100, 200 RU / mL.

Embodiment 2

[0075] Preparation of anti-histone antibody IgG quality control:

[0076] Dilute the anti-histone antibody IgG with the above-mentioned anti-histone antibody IgG calibrator diluent to 20, 100 RU / mL at each concentration point.

Embodiment 3

[0078] Preparation of Reagent No. 1:

[0079] Step 1) Prepare reagent No. 1 dilution:

[0080] Add 800ml of purified water, 12.1g of Tris, 5.8g of sodium chloride and 2ml of Proclin300 into the container, stir well until completely dissolved; add 5g of bovine serum albumin into the container, stir well until completely dissolved; dissolve the solution with 4M HCL Adjust the pH value of the solution to 7.0-7.5; dilute the solution to 1L with purified water, filter it with a 0.2μm filter to obtain the No. 1 dilution of the reagent, and store it at 2-8°C for later use;

[0081] Step 2) Prepare reagent No. 1:

[0082] Dissolve the histone antigen in purified water, dialyze it with carbonate buffer solution with a concentration of 0.2M and a pH of 9.0 at 2-8°C for 2 hours, and then concentrate it to an antigen solution with a concentration of 2-4 mg / mL. Prepare a biotin solution with a concentration of 0.5-1.0mg / ml in 0.2M carbonate buffer solution with a pH of 8.5-9; add it to t...

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Abstract

The invention discloses a magnetic particle-based quantitative chemiluminescent assay kit for an anti-histone antibody IgG. The kit comprises an anti-histone antibody IgG calibrator, an anti-histone antibody IgG quality control product, a Tris buffer containing biotin-labeled histone antigen and bovine serum albumin, a Tris buffer containing an alkaline phosphatase-labeled goat anti-human polyclonal antibody and bovine serum albumin, a Tris buffer containing streptavidin-labeled magnetic particles and bovine serum albumin, and a rinsing solution. The detection method using the kit improves sensitivity and a linearity range by 3 to 5 orders of magnitudes on the basis of a traditional membrane strip immunization method and a traditional enzyme linked immunosorbent assay method, realizes real quantitative determination, is rapid in reaction and reliable in results, can be automatically used in cooperation with an automatic chemiluminescence immunity analyzer and is of irreplaceable important value to clinical diagnosis.

Description

technical field [0001] The invention relates to the technical field of biological detection, in particular to a magnetic particle chemiluminescence quantitative assay kit for anti-histone antibody IgG and a preparation and detection method. Background technique [0002] Histone is a basic nucleoprotein consisting of five subunits (H1, H2A, H2B, H3, H4). Anti-histone antibodies can appear in a variety of connective tissue diseases and are not diagnostically specific, but the positive rate is higher in patients with systemic lupus erythematosus and drug-induced lupus erythematosus. [0003] Anti-histone antibodies are associated with various autoimmune diseases. In patients with systemic lupus erythematosus (SLE), the detection rate of anti-histone antibodies is 30%-70%, and the positive rate in patients with rheumatoid arthritis without complications 15%-50%, the positive rate is 60% in patients with juvenile rheumatoid arthritis, and the detection rate of anti-histone prote...

Claims

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Application Information

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IPC IPC(8): G01N21/76G01N33/532G01N33/535
CPCG01N21/76G01N33/532G01N33/535
Inventor 不公告发明人
Owner 北京贝尔医疗设备有限公司
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