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Magnetic particle chemiluminescence quantitative determination reagent kit of anti-centromere antibody IgG and preparation and detection method

An anti-centromere antibody, chemiluminescence technology, applied in chemiluminescence/bioluminescence, biological testing, analysis by chemical reaction of materials, etc., can solve the problems of low sensitivity, unseen application, narrow linear range, etc.

Inactive Publication Date: 2016-07-20
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.
[0004] At present, the application of magnetic particle chemiluminescence assay in anti-centromere antibody IgG immunoassay products has not yet been seen.

Method used

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  • Magnetic particle chemiluminescence quantitative determination reagent kit of anti-centromere antibody IgG and preparation and detection method
  • Magnetic particle chemiluminescence quantitative determination reagent kit of anti-centromere antibody IgG and preparation and detection method
  • Magnetic particle chemiluminescence quantitative determination reagent kit of anti-centromere antibody IgG and preparation and detection method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0067] Preparation of anti-centromere antibody IgG calibrator:

[0068] Step 1) Prepare anti-centromere antibody IgG calibrator dilution:

[0069] 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 In the container, stir well until it is 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 it with a 0.2μm filter to obtain the anti-centromere antibody IgG calibrator dilution solution, stored at 2-8°C for later use;

[0070] Step 2) Prepare anti-centromere antibody IgG calibrator:

[0071] Dilute anti-centromere antibody IgG with anti-centromere antibody IgG calibrator diluent to each concentration point of 0, 5, 20, 50, 100, 200 RU / mL.

Embodiment 2

[0073] Preparation of anti-centromere antibody IgG quality control:

[0074] Dilute the anti-centromere antibody IgG with the above-mentioned anti-centromere antibody IgG calibrator diluent to 20, 100 RU / mL at each concentration point.

Embodiment 3

[0076] Preparation of Reagent No. 1:

[0077] Step 1) Prepare reagent No. 1 dilution:

[0078] 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; use 4M HCL to dissolve the solution Adjust the pH value 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 until use;

[0079] Step 2) Prepare reagent No. 1:

[0080] Dissolve the centromere antigen in purified water, dialyze with carbonate buffer solution with a concentration of 0.2M and pH 9.0 for 2 hours at 2-8°C, and then concentrate to an antigen solution with a concentration of 2-4mg / mL. Prepare a biotin solution with a concentration of 0.5-1.0mg / ml in a carbonate buffer solution of 0.2M and pH 8.5-9; prepare the biotin solution at the cent...

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Abstract

The invention discloses a magnetic particle chemiluminescence quantitative determination reagent kit of anti-centromere antibody IgG. The reagent kit comprises an anti-centromere antibody IgG calibration material, an anti-centromere antibody IgG quality control product, a Tris buffering solution containing biotin-labeled centromere antigens and bovine serum albumin, a Tris buffering solution containing alkaline phosphatase-labeled goat-anti-human polyclonal antibodies and bovine serum albumin, a Tris buffering solution containing streptavidin-labeled magnetic particles and bovine serum albumin, and a cleaning solution. According to a detection method of the reagent kit, on the basis of a traditional membrane strip immunization method and enzyme linked immunosorbent assay, sensitivity and the linear range are improved by 3-5 magnitude orders, quantitative determination in real sense is achieved, responding is fast, the result is reliable, and the magnetic particle chemiluminescence quantitative determination reagent kit can be full automatically used in cooperation with a full-automatic chemiluminescence immunity analyzer and has irreplaceable important value on 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-centromere antibody IgG and a preparation and detection method. Background technique [0002] Anticentromere antibody (anticentromereantibody, ACA) refers to a class of antibodies that recognize protein antigens present in the centromere region. Currently known CENP-B, CENP-A and CENP-C are the three main autoantigens of ACA. ACA is a biomarker for the diagnosis of some autoimmune diseases, especially for the diagnosis of limited scleroderma (limited cutaneous systemics clerosis, lcssc). At present, ACA is recognized as an important biomarker for the diagnosis and prognosis of systemic scleroderma. [0003] At present, there are membrane strip immunoassay and enzyme-linked immunosorbent assay for clinical detection of anti-centromere antibody IgG. Membrane immunoassay uses membrane strip chromog...

Claims

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Application Information

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IPC IPC(8): G01N21/76G01N33/68
CPCG01N21/76G01N33/68
Inventor 不公告发明人
Owner 北京贝尔医疗设备有限公司
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