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Incomplete antibody detection kit and detection method

A detection kit and complete antibody technology, applied in the field of medical detection, can solve the problems of cumbersome and time-consuming washing process, and achieve the effects of easy automation, improved detection efficiency, and easy interpretation

Inactive Publication Date: 2018-10-19
SUZHOU INST OF BIOMEDICAL ENG & TECH CHINESE ACADEMY OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The washing process is cumbersome and time-consuming, which is limited in clinical application

Method used

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  • Incomplete antibody detection kit and detection method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] A water-based gel-based detection method for incomplete red blood cell antibodies in a microwell plate, the specific steps are:

[0037] (1) Coating antihuman globulin on a U-shaped microwell plate: prepare antihuman globulin to a concentration of 10 μg / mL with pH9.6, 0.05M carbonate buffer, and add to each well of a U-shaped microwell plate 100 μL, put in the refrigerator at 4°C overnight for coating, take it out the next day, wash 3 times with pH 7.2, 0.01M phosphate buffer containing 0.05% Tween, tap dry, and store at 4°C for use;

[0038] (2) Add 200 μL of aqueous gel chromatography medium to the microwell plate, then add the plasma or serum to be tested, then add screening red blood cells, and incubate at 37°C for 30 minutes;

[0039] (3) Carry out centrifugal separation, wherein the rotating speed during centrifugal is 400g, and centrifugal time is 6 minutes;

[0040] (4) Observing the results, the red blood cells spread on the bottom of the U-shaped microwell pl...

Embodiment 2

[0044] A method for detecting incomplete platelet antibody on a microporous plate based on water-based gel, the specific steps are:

[0045] (1) Coating antihuman globulin on a U-shaped microwell plate: prepare antihuman globulin to a concentration of 15 μg / mL with pH9.6, 0.05M carbonate buffer, and add 100 μL, put in the refrigerator at 4°C overnight for coating, take it out the next day, wash 3 times with pH 7.2, 0.01M phosphate buffer containing 0.05% Tween, tap dry, and store at 4°C for use;

[0046] (2) Add 200 μL of aqueous gel chromatography medium to the microwell plate, then add the plasma or serum to be tested, then add platelets and indicator particles, that is, latex particles or magnetic microspheres coated with platelet antibodies, and incubate at 37°C for 30 minutes ;

[0047] (3) centrifugation or magnetic separation;

[0048] (4) Observing the results, it is positive if the indicated particles are spread flat on the bottom of the U-shaped microwell plate, ne...

Embodiment 3

[0050] A method for detecting pathogenic microorganism antibodies on a microporous plate based on water-based gel, the specific steps are:

[0051] (1) Coating antihuman globulin on a U-shaped microwell plate: prepare antihuman globulin to a concentration of 15 μg / mL with pH9.6, 0.05M carbonate buffer, and add 100 μL, put in the refrigerator at 4°C overnight for coating, take it out the next day, wash 3 times with pH 7.2, 0.01M phosphate buffer containing 0.05% Tween, tap dry, and store at 4°C for use;

[0052] (2) Add 200 μL of aqueous gel chromatography medium to the microwell plate, then add plasma or serum to be tested, then add latex particles or magnetic microspheres coated with pathogenic microorganism antigens, and incubate at 37°C for 30 minutes;

[0053] (3) centrifugation or magnetic separation;

[0054] (4) Observing the results, it is positive if the indicated particles are spread flat on the bottom of the U-shaped microwell plate, negative if they gather at the ...

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Abstract

The invention discloses an incomplete antibody detection kit which comprises a U-shaped micro-pore plate, an aqueous glue chromatography medium and granular antigen, wherein the U-shaped micro-pore plate is wrapped with antihuman globulin; the aqueous glue chromatography medium is used for incubating a mixture of serum or plasma with the granular antigen; the granular antigen is reacted with an incomplete antibody in the serum or plasma to form an antigen-antibody compound. The invention further relates to an incomplete antibody detection method. A granular antigen-antibody compound is separated from free antibodies in a centrifugal or magnetic mode, and a tedious washing process in a conventional method can be replaced. When results are observed, if the incomplete antibody is sensitized on the granular antigen, the incomplete antibody can be captured by the antihuman globulin and is spread at the bottom of the U-shaped micro-pore plate, and a positive result is judged; if sensitization is not resulted, the incomplete antibody is not captured but being focused at the bottom to form a small point, and a negative result is judged; a weakly positive result is judged if a situation falls in between. By adopting the method, operation steps are greatly simplified, the detection efficiency can be improved, results are easy to judge, in addition, a great amount of samples can be detected simultaneously, and automation can be achieved easily.

Description

technical field [0001] The invention relates to the field of medical detection, in particular to an incomplete antibody detection kit and detection method. Background technique [0002] Incomplete antibodies refer to antibodies that can bind (sensitize) to corresponding antigen particles in saline medium, but cannot show agglutination reaction visible to the naked eye. Incomplete antibodies are mainly IgG antibodies. After binding to the corresponding antigen of red blood cells (that is, sensitized red blood cells), special methods, such as anti-human globulin method, polybrene method, and enzymatic method, can be used to cause agglutination of sensitized red blood cells. Reaction to detect such antibodies. [0003] Incomplete antibody is the main cause of clinically common delayed transfusion reactions. If there are clinically meaningful antibodies in the blood recipient, the transfused red blood cells containing the corresponding antigens will be destroyed, and hemolytic...

Claims

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Application Information

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IPC IPC(8): G01N33/68G01N33/80
CPCG01N33/6854G01N33/80
Inventor 陈晔洲王红梅李勇段生宝丁少华魏双施蒙青林
Owner SUZHOU INST OF BIOMEDICAL ENG & TECH CHINESE ACADEMY OF SCI
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