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Method and system for detecting whether incomplete antibodies exist on surfaces of red blood cells and application thereof

A complete antibody and red blood cell technology, applied in measuring devices, biological testing, material inspection products, etc., can solve problems affecting the stability of results, sample volume and antibody batch differences, time-consuming and labor-intensive, etc., to achieve clear and easy-to-read reaction results , get rid of the dependence of gel, and avoid the effect of manual operation

Pending Publication Date: 2021-07-13
JIANGYIN LIBO MEDICINE BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, these methods have drawbacks, such as:
[0004] The test tube method requires manual mixing steps, which is time-consuming and laborious. In addition, the test tube method needs to shake the test tube when judging the degree of agglutination. The strength will easily affect the test results. Different people with the same sample may produce different judgment results. Especially for some weak positive samples, it is easy to produce manual errors;
[0005] The sample volume of antibodies and red blood cells in the microplate method and the difference between antibody batches will affect the stability of the results;
[0006] The reagent gel of the micro-column gel method is easy to deform and generate air bubbles during transportation, and the temperature will also affect the size of the gel molecular sieve, thus affecting the stability and reliability of the final test results

Method used

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  • Method and system for detecting whether incomplete antibodies exist on surfaces of red blood cells and application thereof
  • Method and system for detecting whether incomplete antibodies exist on surfaces of red blood cells and application thereof
  • Method and system for detecting whether incomplete antibodies exist on surfaces of red blood cells and application thereof

Examples

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Embodiment 1

[0046] Example 1: A method for detecting whether there are incomplete antibodies on the surface of red blood cells

[0047] This embodiment provides a method for detecting whether there are incomplete antibodies on the surface of red blood cells (see the principle of detection) figure 1 , for possible test results see figure 2 ), the method comprises the steps of:

[0048] (1) Coating: Dilute gamma globulin with a mass volume ratio of 10% by 2048 times with a carbonate buffer solution of pH 9.6 to obtain a dilution; After the well plate was coated, it was coated at 4°C for 12 hours to obtain a coated microwell plate;

[0049] (2) The first plate washing step: Add pH 7.2 PBS buffer solution to the coated microwell plate at an amount of 250 μL per well for washing, repeat washing 5 times, and the microwell plate needs to be washed every time. Blot the remaining liquid in to obtain washed microwell plate A;

[0050] (3) Blocking: After adding 250 μL per hole to the washed mi...

Embodiment 2

[0054] Example 2: A method for detecting whether there are incomplete antibodies on the surface of red blood cells

[0055] This embodiment provides a method for detecting whether there are incomplete antibodies on the surface of red blood cells (see the principle of detection) figure 1 , for possible test results see figure 2 ), the method comprises the steps of:

[0056] (1) Coating: Dilute gamma globulin with a mass volume ratio of 10% by 1024 times with a carbonate buffer solution of pH 8.9 to obtain a dilution; After the well plate was coated, it was coated at 2°C for 8 hours to obtain a coated microwell plate;

[0057] (2) The first plate washing step: Add pH 6.5 PBS buffer solution to the coated microwell plate in an amount of 200 μL per well for washing. Blot the remaining liquid in to obtain washed microwell plate A;

[0058] (3) Blocking: After adding the carbonate buffer of pH 8.9 containing BSA with a volume concentration of 0.1% and Tween20 with a volume conc...

Embodiment 3

[0062] Example 3: A method for detecting whether there are incomplete antibodies on the surface of red blood cells

[0063] This embodiment provides a method for detecting whether there are incomplete antibodies on the surface of red blood cells (see the principle of detection) figure 1 , for possible test results see figure 2 ), the method comprises the steps of:

[0064] (1) Coating: Dilute gamma globulin with a mass volume ratio of 10% by 512 times with a carbonate buffer solution of pH 9.8 to obtain a dilution; After the well plate was coated at 8°C for 16 hours, the coated microwell plate was obtained;

[0065] (2) The first plate washing step: Add pH 7.8 PBS buffer solution to the coated microwell plate in an amount of 300 μL per well for washing, and repeat the washing 5 times. Blot the remaining liquid in to obtain washed microwell plate A;

[0066] (3) Blocking: After adding 300 μL per hole to the washed microwell plate A, the carbonate buffer containing pH 9.8 o...

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Abstract

The invention relates to a method and system for detecting whether incomplete antibodies exist on the surfaces of red blood cells and application thereof, and belongs to the technical field of biological detection. The invention provides a method for detecting whether incomplete antibodies exist on the surfaces of red blood cells based on solid-phase adsorption, which comprises the following steps: adding red blood cells to be detected and anti-human globulin into a container coated with gamma globulin for centrifuging; observing whether to-be-detected red blood cells are adsorbed at the bottom of the container coated with the gamma globulin after the centrifuging is ended; if the to-be-detected red blood cells are adsorbed at the bottom of the container coated with the gamma globulin, determining that incomplete antibodies exist on the surfaces of the to-be-detected red blood cells, and if the to-be-detected red blood cells are not adsorbed at the bottom of the container coated with the gamma globulin, determining that the surfaces of the red blood cells to be detected has no incomplete antibody. The method has lower detection cost on the premise of keeping higher sensitivity and specificity, has the advantage of easiness in automatic operation, and can avoid personal errors in the operation process and improve the working efficiency.

Description

technical field [0001] The invention relates to a method, a system and an application thereof for detecting whether there are incomplete antibodies on the surface of red blood cells, and belongs to the field of biological detection. Background technique [0002] Antiglobulin test (antiglobulin test) is a technique founded by Coombs in 1945 to detect incomplete blood group antibodies other than IgM antibodies on blood recipients or pregnant women, newborn serum (plasma) or red blood cells, also known as Coombs test, among them, the method used to detect other incomplete blood group antibodies except IgM antibody on red blood cells is called direct antiglubin test (Direct antiglubin test, DAT). The direct antihuman sphere test is used to assess the risk of hemolysis caused by incomplete blood group antibodies other than IgM in blood recipients, pregnant women, and fetuses. [0003] At present, the general direct anti-human globulin test is based on the principle of incomplete...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/569G01N33/68
CPCG01N33/80G01N33/6854
Inventor 高明陈玉平陈芳芳王布强徐丹门泉禄徐孟静
Owner JIANGYIN LIBO MEDICINE BIOTECH
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