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Kit for quantitatively determining anti-nRNP/Sm antibody IgG by using magnetic particle chemiluminescence and detection method

A chemiluminescence, quantitative detection technology, applied in the field of immunological detection, can solve the problems of narrow linear range, poor repeatability, low sensitivity, etc.

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

[0005] 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 the 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 for quantitatively determining anti-nRNP/Sm antibody IgG by using magnetic particle chemiluminescence and detection method
  • Kit for quantitatively determining anti-nRNP/Sm antibody IgG by using magnetic particle chemiluminescence and detection method
  • Kit for quantitatively determining anti-nRNP/Sm antibody IgG by using magnetic particle chemiluminescence and detection method

Examples

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Effect test

Embodiment 1

[0066] A kit for quantitatively detecting anti-nRNP / Sm antibody IgG using magnetic particle chemiluminescence, the kit comprising: Anti-nRNP / SmIgG calibrator, Anti-nRNP / SmIgG reagent No. 1, Anti-nRNP / SmIgG reagent No. 2, Anti-nRNP / SmIgG Magnetic Separation Reagent, Anti-nRNP / SmIgG Quality Control and Cleaning Solution.

[0067] The Anti-nRNP / SmIgG calibrator includes 6 liquid calibration bottles with a loading capacity of 0.5mL, and the target concentrations in the liquid calibration bottles are respectively 0, 5, 20, 50, 100, 200RU / mL Anti-nRNP / SmIgG Tris buffer for SmIgG and bovine serum albumin;

[0068] The Anti-nRNP / SmIgG reagent No. 1 is a bottle of 5 mL Tris buffer containing biotin-labeled nRNP / Sm antigen and bovine serum albumin;

[0069] The Anti-nRNP / SmIgG reagent No. 2 is a bottle of 15mL Tris buffer containing alkaline phosphatase-labeled goat anti-human polyclonal antibody and bovine serum albumin;

[0070] The Anti-nRNP / SmIgG magnetic separation reagent is a T...

Embodiment 2

[0117] A preparation method of a kit for quantitative detection of anti-nRNP / Sm antibody IgG using magnetic particle chemiluminescence comprises the following steps:

[0118] Step 1. preparing described Anti-nRNP / SmIgG calibrator comprises: Anti-nRNP / SmIgG calibrator diluent preparation step and Anti-nRNP / SmIgG calibrator preparation step:

[0119] The preparation step of described Anti-nRNP / SmIgG calibrator dilution comprises:

[0120] 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;

[0121] The preparation step of described Anti-nRNP / SmIgG calibrator ...

Embodiment 3

[0139] On the basis of Example 2, the kit also includes a cleaning solution, the preparation steps of the cleaning solution:

[0140] 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, filter with a 0.2μm filter, and store at a temperature range of 2-8°C. Before the experiment, the cleaning solution was diluted by adding purified water to 100 mL for every 7.5 mL and mixed well before use.

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Abstract

The invention relates to the technical field of immunology detection, and concretely relates to a kit for quantitatively determining anti-nRNP / Sm antibody IgG by using magnetic particle chemiluminescence and a detection method. The kit for quantitatively determining anti-nRNP / Sm antibody IgG by using magnetic particle chemiluminescence comprises an anti-nRNP / Sm IgG calibrator, an anti-nRNP / Sm IgG No.1 reagent, an anti-nRNP / Sm IgG No.2 reagent, an anti-nRNP / Sm IgGmagnetic separation reagent, an anti-nRNP / Sm IgG quality-control sample and a cleaning fluid. The invention also discloses preparation and detection methods of the above kit. On the basis of a conventional membrane-strip immunization method and an enzyme linked immunosorbent assay adsorption method, the detection method improves the sensitivity and the linearity range by 3-5 magnitude orders, realizes quantitative detection, possesses the advantages of high sensitivity, strong specificity, good accuracy, low cost, simple operation and visual result determination, is matched with a full-automatic luminescence immunoassay instrument for full-automatic usage, and possesses wide application prospect.

Description

technical field [0001] The invention relates to the technical field of immunological detection, in particular to a kit for quantitatively detecting anti-nRNP / Sm antibody IgG by using magnetic particle chemiluminescence, and a preparation method and a detection method thereof. Background technique [0002] In 1971, Sharp found high titers of anti-ribonucleoprotein antibodies in the serum of a group of patients with special connective tissue diseases, and named this connective tissue disease as mixed connective tissue disease. Mixed connective tissue disease is a combination of systemic lupus erythematosus, polymyositis, scleroderma, and rheumatoid arthritis at the same time or at different times. Patients mainly present with Raynaud's phenomenon, swollen fingers, rash, joints and Lung damage and other diseases. The detection rate of high-titer anti-nRNP / Sm antibody IgG can reach 95-100% in patients with mixed connective tissue disease. Antibody titer is related to disease ac...

Claims

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Application Information

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IPC IPC(8): G01N21/76G01N33/553
Inventor 不公告发明人
Owner 北京贝尔医疗设备有限公司
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