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Magnetic particles chemiluminescence detection method

A chemiluminescence detection and magnetic particle technology, applied in measurement devices, scientific instruments, instruments, etc., can solve the problems of low sensitivity, narrow linear range, and difficult to achieve full automation of enzyme-linked immunosorbent assay, and improve detection accuracy, Good stability and improved detection sensitivity

Inactive Publication Date: 2019-02-15
NINGBO AUCHEER BIOTECHNOLOGY CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Membrane strip method is a qualitative detection method, and enzyme-linked immunoassay method has methodological limitations such as low sensitivity, narrow linear range, and difficulty in realizing full automation.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] A method for detecting chemiluminescence of magnetic particles includes the following steps:

[0020] (1) Add 100ul of reference substance and sample to be tested in the reaction cup, and then add 100ul of fluorescein isothiocyanate-labeled antibody solution. The concentration of fluorescein isothiocyanate-labeled antibody solution is 1ug / ml, 100ul Alkaline phosphatase-labeled antibody solution, the concentration of alkaline phosphatase-labeled antibody solution is 1ug / ml, mix well, incubate at 37℃ for 30min; add 50ul magnetic particle suspension coated with fluorescein isothiocyanate antibody The concentration of the magnetic particle suspension coated with fluorescein isothiocyanate antibody is 1ug / ml, mix well, and incubate at 37°C for 5 min;

[0021] (2) Use a magnetic separator to wash the reaction cup, make the magnetic particles settle in a magnetic field, remove the supernatant, add 300ul of 0.01M PBS buffer containing 0.05% TW-20, pH 7.2, remove the magnetic field, ...

Embodiment 2

[0027] A method for detecting chemiluminescence of magnetic particles includes the following steps:

[0028] (1) In the reaction cup, add 50ul reference substance and sample to be tested in sequence, and then add 50ul containing fluorescein isothiocyanate-labeled antibody solution, the concentration of fluorescein isothiocyanate-labeled antibody solution is 1ug / ml, 50ul Alkaline phosphatase-labeled antibody solution, the concentration of alkaline phosphatase-labeled antibody solution is 1ug / ml, mix well, incubate at 37℃ for 30min; add 50ul magnetic particle suspension coated with fluorescein isothiocyanate antibody The concentration of the magnetic particle suspension coated with fluorescein isothiocyanate antibody is 1ug / ml, mix well, and incubate at 37°C for 5 min;

[0029] (2) Use a magnetic separator to wash the reaction cup, make the magnetic particles settle in the magnetic field, remove the supernatant, add 200ul of 0.01M PBS buffer containing 0.05% TW-20, pH 7.2, remove the...

Embodiment 3

[0035] The preparation method of the antibody solution containing fluorescein isothiocyanate label is: prepare 0.5mg / mL fluorescein isothiocyanate solution with 0.1mol / L pH=9.0 carbonate buffer; according to the ratio of antibody to FITC molecule Add the fluorescein isothiocyanate solution to the antibody solution at a ratio of 1:10, mix well, and let stand at room temperature for 12 hours to react to form an antibody-fluorescein complex; pass the antibody-fluorescein conjugate through the G-25 gel The column is separated to remove the unreacted fluorescein isothiocyanate to obtain a purified antibody-fluorescein complex; the purified antibody-fluorescein complex is used with 0.5% bovine serum albumin pH=8.0 0.1mol / L Tris The buffer is diluted to an antibody-fluorescein complex concentration of 1ug / mL.

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PUM

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Abstract

The invention discloses a magnetic particles chemiluminescence detection method. The method combines with the magnetic separation technology, the chemiluminescence technology, and the immunoassay technology. The method fully utilizes the rapid and easy automation performance of the magnetic separation technology, the high sensitivity performance of the chemiluminescence technology, and the specificity of the immunoassay, and exhibits an irreplaceable role in the field of biological analysis. Compared with a previous membrane strip immunoassay method and enzyme-linked immunosorbent assay method, the method has greatly improved the detection sensitivity, the detection range, the detection time, and the automation operation. According to the magnetic particles chemiluminescence detection method, the antigen-antibody binding surface area is large, and the detection sensitivity is improved by increasing the binding amount of the antigen-antibody; the reaction speed is fast, and the antigen-antibody can be rapidly captured; the combined phase and free phase are easy to separate, which can improve the detection accuracy; and the alkaline phosphatase luminescence system has good stability.

Description

Technical field [0001] The invention relates to the field of in vitro diagnostic reagents, in particular to a magnetic particle chemiluminescence detection method. Background technique [0002] Autoimmune diseases are diseases caused by autoimmune reactions to a certain degree. The most basic function of the immune system is to recognize itself and recognize foreign bodies, so as to protect itself and reject foreign bodies. There may be a variety of antibodies against self-antigens in normal human serum, but their levels are extremely low, not enough to destroy self-components, and can eliminate aging and degenerated self-tissues. This is an autoimmune response. When this reaction is too strong, causing serious tissue damage, and showing clinical symptoms, it is an autoimmune disease. The detection of these indicators can be used as a reference for the diagnosis of diseases, but the detection of a single indicator cannot directly lead to the results of the disease diagnosis. It...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/533G01N33/543
CPCG01N33/533G01N33/54326
Inventor 黄丹娣俞辰泽董文霞周义正
Owner NINGBO AUCHEER BIOTECHNOLOGY CO LTD
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