Method for detecting platelet antibody specificity by using flow cytometry and detection kit

A platelet antigen specific technology, applied in the direction of measuring devices, instruments, scientific instruments, etc., can solve the problem of scarcity of detection methods and achieve the effect of less sample consumption, simple steps and accurate results

Pending Publication Date: 2020-05-19
天津美瑞特医疗科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Furthermore, there are currently few clinical detection methods for antibodies against a specific platelet-specific antigen (HPA) in the body of the person being tested.

Method used

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  • Method for detecting platelet antibody specificity by using flow cytometry and detection kit
  • Method for detecting platelet antibody specificity by using flow cytometry and detection kit
  • Method for detecting platelet antibody specificity by using flow cytometry and detection kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Example 1: Platelet Antibody Specific Detection Kit

[0030] A detection kit for detecting the specificity of platelet antibodies by flow cytometry, comprising platelet antigen-specific fluorescent microspheres coated with different anti-platelet-specific glycoprotein antibodies on the surface of different fluorescent microspheres, and microsphere buffer solution, platelet standards, platelet lysate, calibration beads, and detection antibodies.

Embodiment 2

[0031] Example 2: Preparation of a specific detection kit for platelet antibody detection by flow cytometry.

[0032] This embodiment is the preparation of the platelet antibody specific detection kit, which is carried out according to the following steps:

[0033] 1. Preparation of microsphere buffer

[0034] The microsphere buffer solution involved in this example is a PBS solution containing 5% FBS by volume. NaN3 (w / v), mix well, and filter through 0.45 μm water filter membrane.

[0035] 2. Preparation of platelet antigen-specific fluorescent microspheres

specific Embodiment approach

[0036]All platelet antigen-specific fluorescent microspheres of the present invention are prepared by coating anti-platelet-specific glycoprotein antibodies on carboxylated polystyrene microspheres labeled with APC fluorescein. The method is the same as the reaction system, so this embodiment Take the preparation of anti-CD109 antibody-specific fluorescent microspheres as an example. The specific implementation is as follows:

[0037] 2.1 Solution preparation

[0038] 2.1.1 Preparation of buffer A: Weigh 0.96g NaH2PO4•2H2O, add H2O 600mL to dissolve, adjust the pH value to 6.0, add H2O to the volume to 800 mL, filter with 0.45 μm water filter membrane;

[0039] 2.1.2 Preparation of buffer B: Weigh 7.5g of glycine, add 800mL of H2O to dissolve, adjust the pH value to 7.5, add H2O to the volume to 1000mL, and filter with a 0.45 μm water filter membrane;

[0040] 2.1.3 Preparation of 50 mg / mL 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC) solution: Weigh 1 g ...

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Abstract

The invention provides a method for detecting platelet antibody specificity by using flow cytometry. The method comprises the following steps: (1) preparing platelet antigen specific fluorescent microspheres; (2) preparing a detection antibody; (3) preparing a platelet reference substance; (4) detecting a reaction system; and (5) detecting a sample to be detected. The invention also aims at providing a detection kit for detecting the specificity of the platelet antibody by applying the flow cytometry. The kit comprises platelet antigen specific fluorescent microspheres obtained by coating thesurfaces of different fluorescent microspheres with different anti-platelet specific glycoprotein antibodies, a microsphere buffer solution, a platelet reference substance, a platelet lysate, a cleaning solution, calibration microspheres and a detection antibody. The invention discloses a platelet specific antibody detection method. In the prior art, the platelet antibody detection mainly aims atthe platelet antibody of the detected person; compared with the prior art, the method has the advantages that the problem of free platelet antibodies in blood plasma or blood serum is solved, the typeof the platelet specific antigen (HPA) of the antibody existing in a detected person can be further determined, and the method is small in sample use amount, more accurate in result, simple in step,easy to operate and higher in efficiency.

Description

technical field [0001] The invention belongs to the field of immunodiagnosis and detection, and in particular relates to a method and a detection kit for detecting platelet antibody specificity by using flow cytometry. Background technique [0002] Platelet antibodies are pathogenic antibodies produced by patients exposed to foreign platelet antigens during blood transfusion, pregnancy or transplantation, which can sensitize and destroy platelets, causing thrombocytopenia, leading to platelet transfusion refractoriness (plalelet transfusion refractoriness) , PTR), fetal-maternal alloimmune thrombocytopenia (FMAIT), posttransfusion purpura (posttransfusion purpura, PTP), and transplantation-associated alloimmune thrombocytopenia (transplantation-associated alloimmune thrombocytopenia , TAAT) and other diseases, patients are prone to symptoms such as petechiae, ecchymosis and bleeding, and even death in severe cases. Patients with immune diseases can also produce antibodies a...

Claims

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Application Information

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IPC IPC(8): G01N33/533G01N33/543
CPCG01N33/533G01N33/54313
Inventor 洪建
Owner 天津美瑞特医疗科技有限公司
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