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Method for detecting double-chain DNA resistant antibody using gold magnetic particle as carrier

A gold magnetic particle and antibody technology, which is applied in the field of detecting anti-double-stranded DNA antibodies in the tested sample by enzyme-linked immunosorbent assay, can solve the problems of poor sensitivity of anti-double-stranded DNA antibodies, etc., and achieves high coating efficiency and large size. The effect of immobilization capacity, sensitivity and specificity improvement

Active Publication Date: 2012-09-19
XIAN GOLDMAG NANOBIOTECH
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Problems solved by technology

[0018] The object of the present invention is to provide a method for detecting anti-double-stranded DNA antibodies, which solves the shortcomings of the poor sensitivity of existing detection anti-double-stranded DNA antibodies

Method used

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  • Method for detecting double-chain DNA resistant antibody using gold magnetic particle as carrier
  • Method for detecting double-chain DNA resistant antibody using gold magnetic particle as carrier

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Experimental program
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Embodiment 1

[0069] Embodiment 1, detect anti-double-stranded DNA antibody on the surface of gold magnetic particles coated with double-stranded DNA

[0070] 1] Immobilization of double-stranded DNA on the surface of gold magnetic particles and sealing and preservation of gold magnetic particles: Take 1 mg of gold magnetic particles in a 2 mL centrifuge tube, magnetically separate, and discard the supernatant. Wash the gold magnetic particles with a coupling buffer (a mixed solution of 20% PEG8000 and 2.5M sodium chloride) until the preservation environment of the gold magnetic particles reaches the pH or salt concentration of the coupling buffer, and each coupling buffer washes The dosage is 400 μL. After the supernatant was discarded by magnetic separation, 14 μg of double-stranded DNA and 200 μL of the above-mentioned coupling buffer were added to the gold magnetic particles, and reacted at 37° C. and 180 rpm for 30 minutes. After discarding the supernatant by magnetic separation, add ...

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Abstract

The method comprises: 1) the GoldMag nano-particles are mixed with the double strand DNA; under certain condition, the double strand DNA is fixed on the surface of GoldMag nano-particles, or the GoldMag nano-particles are used to extract the DNA from the human blood, saliva and semen and to fix the extracted DNA on the surface of GoldMag nano-particles; 2) using indirect ELISA method, immumofluorescence method or chemiluminescent method to detect the anti double strand DNA antibody existed in the sampler under test.

Description

technical field [0001] The invention relates to a method for detecting anti-double-strand DNA antibodies in a test sample, in particular to a method for detecting anti-double-strand DNA antibodies in a test sample by using an enzyme-linked immunosorbent method. Background technique [0002] Systemic lupus erythematosus (SLE) is a relatively common autoimmune disease clinically. The prominent manifestation of patients is the presence of various autoantibodies, the most important of which is anti-double-stranded DNA antibody. This is an autoantibody that binds to natural DNA and is detected almost exclusively in SLE patients. The rise and fall of anti-double-stranded DNA antibody titer is related to the condition of SLE patients. With the control of disease activity, the titer of anti-double-stranded DNA antibody can decrease or disappear. [0003] Early methods such as indirect hemagglutination, complement fixation test and convective immunoelectrophoresis were eliminated du...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/553C12Q1/68
Inventor 崔亚丽陈超李芳
Owner XIAN GOLDMAG NANOBIOTECH
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