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Kit for quantitative detection of anti-M2-3E antibody IgG by using magnetic particle chemiluminescence, and preparation method and detection method thereof

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

Inactive Publication Date: 2016-04-06
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

[0004] 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 quantitative detection of anti-M2-3E antibody IgG by using magnetic particle chemiluminescence, and preparation method and detection method thereof
  • Kit for quantitative detection of anti-M2-3E antibody IgG by using magnetic particle chemiluminescence, and preparation method and detection method thereof
  • Kit for quantitative detection of anti-M2-3E antibody IgG by using magnetic particle chemiluminescence, and preparation method and detection method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0065] A kit for quantitatively detecting anti-M2-3E antibody IgG using magnetic particle chemiluminescence, the kit comprising: Anti-M2-3EIgG calibrator, Anti-M2-3EIgG reagent No. 1, Anti-M2-3EIgG reagent No. 2, Anti-M2-3EIgG Magnetic Separation Reagent, Anti-M2-3EIgG Quality Control and Cleaning Solution.

[0066] The Anti-M2-3EIgG 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-M2- Tris buffer of 3E IgG and bovine serum albumin;

[0067] The Anti-M2-3EIgG reagent No. 1 is a bottle containing 5 mL of Tris buffer containing biotin-labeled M2-3E antigen and bovine serum albumin;

[0068] The Anti-M2-3EIgG reagent No. 2 is a bottle of 15mL Tris buffer containing alkaline phosphatase-labeled goat anti-human polyclonal antibody and bovine serum albumin;

[0069] The Anti-M2-3EIgG magnetic separation reagent is a Tris buffer bottle conta...

Embodiment 2

[0116] A preparation method of a kit for quantitative detection of anti-M2-3E antibody IgG using magnetic particle chemiluminescence comprises the following steps:

[0117] Step 1. preparing described Anti-M2-3EIgG calibrator comprises: Anti-M2-3EIgG calibrator diluent preparation step and Anti-M2-3EIgG calibrator preparation step:

[0118] The preparation step of described Anti-M2-3EIgG calibrator dilution comprises:

[0119] 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;

[0120] The preparation steps of described Anti-M2-3EIgG calibrator comprises: ...

Embodiment 3

[0138] On the basis of Example 2, the kit also includes a cleaning solution, the preparation steps of the cleaning solution:

[0139] 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 particularly relates to a kit for quantitative detection of an anti-M2-3E antibody IgG by using magnetic particle chemiluminescence, and a preparation method and a detection method thereof. The kit for quantitative detection of the anti-M2-3E antibody IgG by using magnetic particle chemiluminescence includes an Anti-M2-3E IgG calibrator, an Anti-M2-3E IgG reagent No. 1, an Anti-M2-3E IgG reagent No. 2, an Anti-M2-3E IgG magnetic separation reagent, an Anti-M2-3E IgG control material and a cleaning solution. The invention also discloses a preparation method and a detection method of the above kit. Based on the traditional film strip immunoassay and enzyme-linked immunosorbent assay, the detection method increases the sensitivity and linearity range by 3-5 orders of magnitude, achieves the quantitative detection, and has the advantages of high sensitivity and specificity, good accuracy, low cost, simple operation and the objective judgment of results. The method cooperates with an automated chemiluminescence immunoassay analyzer to reach fully automation, and has broad application prospects.

Description

technical field [0001] The invention relates to the technical field of immunological detection, in particular to a kit for quantitative detection of anti-M2-3E antibody IgG by using magnetic particle chemiluminescence and a preparation method and detection method thereof. Background technique [0002] Anti-M2-3E antibody IgG is a serological marker for the diagnosis of primary biliary cirrhosis (PBC), which has high sensitivity and specificity. It is also suitable for those who lack obvious abnormal liver function or symptoms of cholestasis in the early stage. At present, there are membrane strip immunoassay and enzyme-linked immunosorbent assay for clinical detection of anti-M2-3E antibody IgG. 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 judgme...

Claims

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Application Information

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IPC IPC(8): G01N33/68G01N33/543
CPCG01N33/6854G01N33/54326G01N33/6893G01N2800/085
Inventor 不公告发明人
Owner 北京贝尔医疗设备有限公司
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