Blood platelet magnetizing and immunolabeling analysis method

An analysis method and immunolabeling technology, applied in the direction of biological testing, material inspection products, etc., can solve the problems of lack of platelet antibody detection and difficulty in being widely used in clinical practice, and achieve easy automation, reduce the process of sealing and washing, and overcome non-specific The effect of heteroadsorption

Inactive Publication Date: 2013-09-25
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

[0006] Other platelet antibody detection methods include: flow cytometry, modified antigen capture ELISA, etc. These methods have their own advantages and disadvantages, and are difficult to be widely used in clinical practice
Therefore, so far, there is no "gold standard" method for platelet antibody detection in the world.

Method used

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  • Blood platelet magnetizing and immunolabeling analysis method

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Comparison scheme
Effect test

Embodiment 1

[0023] The present invention provides a platelet magnetization immune labeling analysis method, which can be used for platelet antibody screening, comprising the following steps:

[0024] (1) Preparation of magnetic beads

[0025] 0.85g, 3.1 mmol of FeCl 3 6H 2 O, 0.30g FeCl 2 4H 2 O was dissolved in 200 mL of water under nitrogen protection. Add an appropriate amount of surfactant, and slowly add 1.5 mol / L ammonia solution to the above solution under strong stirring. When the pH of the solution rises to 6~7, a large amount of black Fe will be produced in the solution. 3 o 4 Particles; continue to add ammonia water to pH = 8, so that the hydrolysis is complete. Aged at 80° C. for 0.5 hour. The resulting solid was separated, washed 3 times with distilled water, and then dispersed in 100 mL of distilled water under the action of ultrasound to obtain Fe 3 o 4 The colloidal solution was collected for later use.

[0026] (2) Sample collection and processing

[0027] 1) N...

Embodiment 2

[0040] The invention provides a platelet magnetization immune labeling analysis method, which can be used for platelet cross-matching, comprising the steps of:

[0041] (1) Preparation of magnetic beads

[0042] 0.85g, 3.1 mmol of FeCl 3 6H 2 O, 0.30g FeCl 2 4H 2 O was dissolved in 200 mL of water under nitrogen protection. Add an appropriate amount of surfactant, and slowly add 1.5 mol / L ammonia solution to the above solution under strong stirring. When the pH of the solution rises to 6~7, a large amount of black Fe will be produced in the solution. 3 o 4 Particles; continue to add ammonia water to pH = 8, so that the hydrolysis is complete. Aged at 80° C. for 0.5 hour. The resulting solid was separated, washed 3 times with distilled water, and then dispersed in 100 mL of distilled water under the action of ultrasound to obtain Fe 3 o 4 The colloidal solution was collected for later use.

[0043] (2) Sample collection and processing

[0044] 1) Donor platelets: a: ...

Embodiment 3

[0057] The invention provides a platelet magnetization immune labeling analysis method, which can be used for platelet antigen typing, comprising the steps of:

[0058] (1) Preparation of magnetic beads

[0059] 0.85g, 3.1 mmol of FeCl 3 6H 2 O, 0.30g FeCl 2 4H 2 O was dissolved in 200 mL of water under nitrogen protection. Add an appropriate amount of surfactant, and slowly add 1.5 mol / L ammonia solution to the above solution under strong stirring. When the pH of the solution rises to 6~7, a large amount of black Fe will be produced in the solution. 3 o 4 Particles; continue to add ammonia water to pH = 8, so that the hydrolysis is complete. Aged at 80° C. for 0.5 hour. The resulting solid was separated, washed 3 times with distilled water, and then dispersed in 100 mL of distilled water under the action of ultrasound to obtain Fe 3 o 4 The colloidal solution was collected for later use.

[0060] (2) Sample collection and processing

[0061] 1) Platelets to be test...

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Abstract

The invention discloses a blood platelet magnetizing and immunolabeling analysis method. The method comprises the following steps: uniformly mixing blood platelets and magnetic beads; magnetizing the blood platelets to obtain suspension; uniformly mixing, incubating and washing the magnetized blood platelets and a detected sample; adding a labeled second antibody and incubating to obtain mixed liquid; applying magnetic force to layer the mixed liquid; determining upper reaction liquid or determining after washing the magnetized blood platelets in the lower layer to obtain a result, namely performing double-phase complementary immune analysis. In the mode, the blood platelet magnetizing and immunolabeling analysis method provided by the invention is used for detecting blood platelet related antigen antibody and cross-matching of blood; the magnetic force is applied so that the magnetized blood platelet is washed without centrifuging; the method is easy and convenient to operate; by a labeled antibody adsorption test for directly determining the upper reaction liquid, the non-specific adsorption in an immunolabeling technology is overcome, the processes of closing and washing are reduced, and the time consumption is short. The method can detect a large number of samples and facilitates automation; the two detection results of the double phases can be verified mutually, so that the accuracy of the detection results is improved.

Description

technical field [0001] The invention relates to a method for detecting platelet antigen antibody and cross-matching type, in particular to a platelet magnetization immunolabeling analysis method. Background technique [0002] Platelet antibodies are produced by allogeneic or autologous platelet antigens acting on the body, including alloantibodies, autoantibodies, alloantibodies, drug-dependent antibodies, and nonspecific antibodies. These antibodies act on human platelets and can lead to platelet destruction And cause maternal alloimmune thrombocytopenia, essential thrombocytopenia, platelet transfusion ineffectiveness, posttransfusion purpura and other platelet immune diseases. Platelet transfusion is an effective measure to treat various bleeding diseases caused by thrombocytopenia. With the increase of platelet transfusion volume and transfusion times, the production of immune-related platelet antibodies in patients increases accordingly. In order to prevent the producti...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/53
Inventor 王红梅李勇段生宝丁少华田晶晶陈烨洲史素霞李冬
Owner SUZHOU INST OF BIOMEDICAL ENG & TECH CHINESE ACADEMY OF SCI
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