Kit for quantitatively detecting anti-SS-A antibody IgG by utilizing magnetic particle chemiluminescence, preparation method and detection method thereof

A chemiluminescent, quantitative detection technology, applied in the field of immunological detection, can solve the problems of long reaction time, poor repeatability, narrow linear range, etc.

Inactive Publication Date: 2016-03-30
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 detecting anti-SS-A antibody IgG by utilizing magnetic particle chemiluminescence, preparation method and detection method thereof
  • Kit for quantitatively detecting anti-SS-A antibody IgG by utilizing magnetic particle chemiluminescence, preparation method and detection method thereof
  • Kit for quantitatively detecting anti-SS-A antibody IgG by utilizing magnetic particle chemiluminescence, preparation method and detection method thereof

Examples

Experimental program
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Effect test

Embodiment 1

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

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

[0068] The Anti-SS-AIgG reagent No. 1 is a bottle of Tris buffer containing biotin-labeled Anti-SS-A antigen and bovine serum albumin with a capacity of 5 mL;

[0069] The Anti-SS-AIgG 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-SS-AIgG magnetic separation reagent is a Tris buffer bottle conta...

Embodiment 2

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

[0118] Step 1. preparing described Anti-SS-AIgG calibrator comprises: Anti-SS-AIgG calibrator diluent preparation step and Anti-SS-AIgG calibrator preparation step:

[0119] The preparation step of described Anti-SS-AIgG 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-SS-AIgG calibrator comprises:

[012...

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 immunological detection, specifically relates to a kit for quantitatively detecting anti-SS-A antibody IgG by utilizing magnetic particle chemiluminescence, a preparation method and a detection method thereof. The kit for quantitatively detecting anti-SS-A antibody IgG by utilizing magnetic particle chemiluminescence comprises an Anti-SS-A IgG calibration product, an Anti-SS-A IgG reagent 1#, an Anti-SS-A IgG reagent 2#, an Anti-SS-A IgG magnetic separation reagent, an Anti-SS-A IgG quality control material and a cleaning fluid. The invention also discloses the preparation method and the detection method for the kit. According to the detection method, on the basis of the traditional film strip immunization and enzyme linked immunosorbent assay, the sensitivity and the linearity range are increased by 3-5 orders of magnitudes and the quantitative detection is realized; the kit has the advantages of high sensitiveness, strong specificity, high accuracy, low cost, convenience in operation and objective judging result; the full-automatic chemiluminiscence immune analyzer is used for realizing the full-automatic use and has wide application prospect.

Description

technical field [0001] The invention relates to the technical field of immunological detection, in particular to a kit for quantitative detection of anti-SS-A antibody IgG by using magnetic particle chemiluminescence and a preparation method and detection method thereof. Background technique [0002] Anti-SS-A antibodies are associated with various autoimmune diseases, and are common in patients with Sjogren's syndrome, systemic lupus erythematosus, and chronic active hepatitis. In addition, anti-SS-A antibodies can also be detected in 100% of neonatal lupus erythematosus, and anti-SS-A antibodies can be transmitted to the fetus through the placenta, causing inflammatory reactions and congenital heart block in newborns. The positive rate of SS-A antibody in other diseases and normal human serum is very low, and the antibody titer is also low, and it is easy to identify in combination with clinical manifestations. [0003] At present, there are membrane strip immunoassay and...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/564G01N33/531
CPCG01N33/564G01N33/531
Inventor 不公告发明人
Owner 北京贝尔医疗设备有限公司
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