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A kit for detecting blood group antibodies and its application

A blood group antibody and kit technology, applied in the field of blood group antibody detection and identification, can solve the problems of only qualitative or semi-quantitative detection, inability to achieve standardization of detection, and complicated and complicated detection methods.

Active Publication Date: 2018-06-08
GENERAL HOSPITAL OF PLA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] An object of the present invention is to provide a blood group antibody identification and / or detection method, which solves the defect that the existing detection method can only qualitatively or semi-quantitatively detect blood group antibodies in blood, and realizes the detection of blood group antibodies, especially blood group antibodies. Quantitative identification and / or detection of weak antibodies, improving the safety of blood transfusion
[0009] Another object of the present invention is to provide a blood group antibody identification and / or detection method, which solves the problem of cumbersome and complicated operations in the existing detection methods, and can realize the automation and batch production of irregular antibodies with ten lines and above detection
[0010] Another object of the present invention is to provide a kit for blood type antibody identification and / or detection, which solves the problems of too short storage period of red blood cells, poor detection reproducibility, and inability to achieve standardization of detection in existing detection products

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Example 1 Preparation of microwell plate coated with ABO antigen

[0037] (1) Add 0.1g / L L-polylysine to the microwell plate, the amount of addition is 100μL / well, put it in a water bath at 37°C for 4h, discard the liquid in the well, wash with distilled water 3 times, 5min / time, and place at 37°C dried and stored for later use;

[0038] (2) use 60 Co irradiated polystyrene microporous plate with an irradiation dose of 8kGy;

[0039] (3) Take 2 mL of fresh type A venous blood, centrifuge at 3000 rpm for 5 minutes to remove the supernatant, and take packed red blood cells; add 20 mL of 5% sodium dodecyl sulfate to the packed red blood cells, leave it at room temperature for 3 minutes, and then centrifuge at 3000 rpm to remove Supernatant; add 20mL pure water to the precipitate, shake and mix well, centrifuge again at 3000rpm to remove the supernatant, repeat 5 times, the obtained precipitate is the red blood cell membrane protein extract containing the A antigen;

[0...

Embodiment 2

[0041] Example 2 Preparation of ABO-type antigen-coated microwell plates

[0042] (1) Add 0.5g / L L-polylysine to the microwell plate, the amount of addition is 200μL / well, put it in a water bath at 37°C for 4h, discard the liquid in the well, wash with distilled water 3 times, 5min / time, and place at 37°C dried and stored for later use;

[0043] (2) use 60 Co irradiated polystyrene microporous plate with an irradiation dose of 8kGy;

[0044] (3) Take 2 mL of fresh B-type venous blood, centrifuge at 3000 rpm for 5 minutes to remove the supernatant, and take packed red blood cells; add 20 mL of 5% sodium dodecyl sulfate to the packed red blood cells, leave it at room temperature for 3 minutes, and then centrifuge at 3000 rpm to remove Supernatant; add 20mL of pure water to the precipitate, shake and mix well, centrifuge again at 3000rpm to remove the supernatant, repeat 5 times, the obtained precipitate is the erythrocyte membrane protein extract containing B antigen;

[0045...

Embodiment 3

[0046] Example 3 Preparation of ABO-type antigen-coated microwell plates

[0047] (1) Add 0.1g / L L-polylysine to the microwell plate, the amount of addition is 100μL / well, put it in a water bath at 37°C for 4h, discard the liquid in the well, wash with distilled water 3 times, 5min / time, and place at 37°C dried and stored for later use;

[0048] (2) use 60 Co irradiated polystyrene microporous plate with an irradiation dose of 8kGy;

[0049](3) Take 2 mL of fresh type A venous blood, centrifuge at 3000 rpm for 5 minutes to remove the supernatant, and take packed red blood cells; add 20 mL of 5% sodium dodecyl sulfate to the packed red blood cells, leave it at room temperature for 3 minutes, and then centrifuge at 3000 rpm to remove Supernatant; add 20mL pure water to the precipitate, shake and mix well, centrifuge again at 3000rpm to remove the supernatant, repeat 5 times, the obtained precipitate is the red blood cell membrane protein extract containing the A antigen;

[0...

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Abstract

The invention provides a kit used for blood group antibody authentication and / or detection and a method for carrying out blood group antibody authentication and / or detection through the kit. The kit comprises a solid phase carrier where blood group antigens are fixed, complements, anticomplements labeled through enzymes and color developing liquid. The kit used for blood group antibody authentication and / or detection is long in storage period and convenient to store, and the blood group antibody detection convenience is improved; meanwhile, by means of the method for blood group antibody authentication and / or detection, antibodies in blood can be quantitatively detected, detection of weak antibodies in the blood is achieved, personal errors possibly caused by naked eye judgment in an existing detection method are avoided, detection accuracy and detection reproducibility are improved, and blood transfusion safety is guaranteed; in addition, it can be achieved that automatic and batch detection of irregular antibodies can be carried out through cells in the tenth lineage or the higher lineage.

Description

technical field [0001] The invention belongs to the technical field of blood type antibody detection and identification, and in particular relates to a kit for blood type antibody detection and its application in blood type antibody detection. Background technique [0002] Blood typing is very important in blood transfusion therapy, which is based on cell agglutination caused by antigen-antibody reaction. Detection of blood group antibodies in serum / plasma using RBC surface antigens is called reverse typing. In clinical blood transfusion, it is necessary to ensure that the identification results of positive and negative stereotyping methods are consistent in order to ensure the safety of blood transfusion. At present, most of the anti-typing tests use the active erythrocyte surface antigen as the standard antigen. However, fresh erythrocytes are not easy to store, and the antigenicity of the erythrocyte surface antigen gradually decreases with the prolongation of storage ti...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/80
CPCG01N33/80
Inventor 汪德清杨璐
Owner GENERAL HOSPITAL OF PLA
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