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Reagent kit for auxiliary diagnosis of autoimmune disease through glucosan specificity antibody

A technology for auxiliary diagnosis and kit, applied in the field of kits for auxiliary diagnosis of autoimmune diseases through dextran-specific antibodies

Inactive Publication Date: 2012-11-07
PEKING UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

For example, for systemic lupus erythematosus and rheumatoid arthritis, currently commonly used laboratory diagnostic indicators are antinuclear antibodies and rheumatoid factor, but they still have a high rate of missed detection and false detection, so looking for and identifying new It is important to use laboratory indicators to diagnose or assist in the diagnosis of autoimmune diseases

Method used

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  • Reagent kit for auxiliary diagnosis of autoimmune disease through glucosan specificity antibody
  • Reagent kit for auxiliary diagnosis of autoimmune disease through glucosan specificity antibody
  • Reagent kit for auxiliary diagnosis of autoimmune disease through glucosan specificity antibody

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] Astragalus polysaccharide (APS): purchased from Pharmagenesis, composed of D-glucose, the main chain is connected by α-1,4-glycosidic bonds, and the side chains are connected by α-1,6-glycosidic bonds.

[0039] Horseradish peroxidase (Horseradish peroxidase, HRP) labeled goat anti-human IgG: purchased from SBA Company;

[0040] Bovine serum albumin (BSA): purchased from Hyclone Company;

[0041] PBS buffer: the concentration of PBS buffer is 0.15mol / L, and the pH is 7.2;

[0042] PBST solution: add Tween 20 to the above PBS buffer, the volume ratio of Tween 20 and PBS buffer is 0.05%;

[0043] PBST-BSA solution: add bovine serum albumin (BSA) to the above-mentioned PBST solution, and the final concentration of bovine serum albumin is 1 gram / L;

[0044] OPD-phosphoric acid citrate buffer (pH 9.6): add o-phenylenediamine in the phosphoric acid citrate buffer, the mass percent concentration of o-phenylenediamine is 0.06; the pH of the phosphoric acid citrate buffer is 5....

Embodiment 2

[0079] 2. Application of kit

[0080] Use the kit prepared in step 1 to assist in the detection of RA disease.

[0081] 1. Serum collection

[0082] Sera were collected from the following 3 groups of people:

[0083] The first group (RA): 30 confirmed cases of RA patients who meet the diagnostic criteria of the American Rheumatology Association and are active patients;

[0084] The second group (HS): 30 cases of healthy people were randomly selected as negative controls;

[0085] The third group (CA): 30 diagnosed cancer patients, including lung cancer patients (7 patients), leukemia patients (4 patients), cervical cancer patients (6 patients), ovarian cancer patients (7 patients), endometrial cancer patients Patients (6 people).

[0086] 2. Detection

[0087] APS was used to coat the ELISA plate, and the APS-specific antibody IgG in the sera of the three groups of people were detected respectively. The specific method is as follows:

[0088]1) Coat 96-well ELISA plate ...

Embodiment 3

[0111] Horseradish peroxidase (Horseradish peroxidase, HRP) labeled goat anti-human IgG: purchased from SBA Company;

[0112] Bovine serum albumin (BSA): purchased from Hyclone Company;

[0113] PBS buffer: the concentration of PBS buffer is 0.15mol / L, and the pH is 7.2;

[0114] PBS-T solution: add Tween 20 to the above PBS buffer, the volume ratio of Tween 20 and PBS buffer is 0.05%;

[0115] PBST-BSA solution: add bovine serum albumin (BSA) to the above-mentioned PBST solution, and the final concentration of bovine serum albumin is 1 gram / L;

[0116] OPD-phosphoric acid citrate buffer (pH 9.6): O-phenylenediamine is added to the phosphoric acid citrate buffer, and the mass percentage concentration of o-phenylenediamine is 0.06%; the pH of the phosphoric acid citrate buffer is 5.0, and the phosphoric acid The concentration is 0.1mol / L, and the concentration of citric acid is 0.05mol / L;

[0117] sulfuric acid;

[0118] 96-well ELISA plate: Costar Company.

[0119] 2. App...

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Abstract

The invention discloses a kit used for helping to diagnose autoimmune diseases by a dextran specific antibody. Both an ELISA kit used for helping to diagnose systemic lupus erythematosus and an ELISA kit used for helping to diagnose rheumatoid arthritis which are discloses by the invention comprise antigens used for coating; the antigen used for coating can be Alpha-1 and 4-dextran, and astragalus polysaccharide is preferable; and the antigen used for coating can also be Alpha-1 and 6-dextran, and dextran is preferable. The kit can be used for helping to diagnose the systemic lupus erythematosus and the rheumatoid arthritis and has important application value.

Description

technical field [0001] The invention relates to a kit for assisted diagnosis of autoimmune diseases by using dextran-specific antibodies. Background technique [0002] The most basic function of the immune system is to identify self and non-self, and then achieve the purpose of protecting itself from external pathogenic microorganisms. Autoimmunity is the ability of the body's immune system to mount an immune response against its own components and occurs in all individuals. Under normal circumstances, the autoimmune response does not cause pathological damage; however, under certain conditions, the immune system still produces an excessive immune response to its own cells or tissue components, resulting in inflammatory damage to its own tissues or organs. A disease state caused by an exaggerated immune response to a component is called an autoimmune disease. Autoimmune diseases can be divided into organ-specific autoimmune diseases and systemic autoimmune diseases. Compa...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/53
Inventor 高晓明戴慧董红亮龙锴张岩徐晓军
Owner PEKING UNIV