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Anti-IgA antibody detection kit

A technology of antibody detection and kit, applied in the field of medical devices, to reduce the risk of blood transfusion and improve the safety of blood transfusion

Active Publication Date: 2015-02-18
SHANGHAI BLOOD CENT
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, this patent is used to detect whether there is a lack of IgA antigen in the serum of patients or blood donors, and there is no report on an anti-IgA antibody detection kit for detecting whether anti-IgA antibodies are contained in the serum of patients or blood donors

Method used

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  • Anti-IgA antibody detection kit

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] The preparation method of embodiment 1 IgA antigen standard substance (coated antigen)

[0044] 1. Centrifuge normal human serum at rpm20000-30000 for 1-2h at 4°C, dilute it with 0.01M-0.05M pH 7.2-PH7.4 PBS for 1-2 times, and then perform affinity chromatography through protein G affinity column. 0.01M-0.05M pH 7.2-PH7.4 PBS buffer equilibrates about 5-10 times the column bed volume, and most IgG is adsorbed on the column.

[0045] 2. Slowly load the sample at a flow rate of 0.3ml-0.8ml / min, collect the passing liquid, and perform protein A affinity chromatography, and equilibrate the column with 0.01M-0.05M pH 7.2-PH7.4 PBS buffer for about 5-10 times bed volume. Constant flow pump controls the elution flow rate, slowly 0.1M-0.5M pH 2.0-PH4.0 glycine-hydrochloric acid buffer elution 5-10 times the volume of the column bed to obtain an eluate rich in IgA, which may also contain a small amount of other antibodies Components (a small amount of IgM and IgG, etc.). Coll...

Embodiment 2

[0049] Experimental steps:

[0050] 1. Dilute the IgA antigen standard 100-1000 times with coating buffer, and add 100ul of diluted IgA to each well. Incubate overnight at 4°C.

[0051] 2. Wash the plate 4 times with plate washer. Add blocking solution 200ul / well after buckling dry. After 30 minutes in a water bath at 37°C, the plate was washed three times and dried, and stored at a low temperature of 4°C. Add 100ul of diluted anti-IgA (+) standard, positive control, negative control and test serum to each well of the flat bottom plate, and bathe in water at 37°C for 1 hour. The serum to be tested was repeated in 4 wells, and the control was repeated in 2 wells.

[0052] 3. Dry the liquid in the wells, wash the plate 5 times with the plate washing solution, and then dry it. Add 100ul of the enzyme-labeled secondary antibody diluted 100-1000 times with the diluent to each well, and place in a water bath at 37°C for 1 hour.

[0053] 4. Dry the liquid in the well, wash the p...

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Abstract

The invention relates to an anti-IgA antibody detection kit. The anti-IgA antibody detection kit consists of a reagent and a kit body, wherein the reagent is placed in the kit body and consists of a sample diluent, a coating buffer solution, a plate washing solution, a confining liquid, a developing solution, a stop solution, a coating antigen, a second enzyme-labeled antibody, an anti-IgA(+) standard sample, a positive control group and a negative control group. The anti-IgA antibody detection kit has the advantages of being capable of being used for routine blood detection in clinical medical treatment units, blood centers and central blood stations to detect whether a blood sample contains the IgA antibody or not; and the anti-IgA antibody detection kit can be used for providing the blood transfusion service for patients, and providing suitable blood products for coordinative infusion so as to improve the blood transfusion safety and reduce the blood transfusion risk.

Description

technical field [0001] The present invention relates to the field of medical devices, in particular to anti-IgA antibody detection technology, in particular to an anti-IgA kit for screening IgA-deficiency patients and patients with autoimmune diseases. Background technique [0002] In the prior art, there are methods for detecting anti-IgA antibodies using radioimmunoassay technology and gel card technology. These methods have high requirements for laboratories, have radioactive pollution, are complicated to operate, require special supporting instruments, are expensive, and are not suitable for Blood donors and patients are screened and tested, which is not conducive to popularization and application. [0003] In the prior art, there are technical problems such as long detection time of anti-IgA antibody, high laboratory requirements, and danger to detection personnel. [0004] IgA is an immunoglobulin, which accounts for 10-15% of total serum immunoglobulins. According t...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/53
Inventor 陆萍凌冰李睿书刘达庄
Owner SHANGHAI BLOOD CENT
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