Flow micro-sphere method for detecting plastocyte specificity immune body

A technology of flow microspheres and platelets, which is applied in the direction of measuring devices, instruments, scientific instruments, etc., can solve the problems of limited application, high specificity, and low sensitivity, and achieve the effects of convenient operation, high sensitivity, and mature technology

Inactive Publication Date: 2008-08-20
侯明 +2
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Problems solved by technology

These methods have high specificity and low sensitivity, which l

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Embodiment Construction

[0011] Detection steps of the present invention are as follows:

[0012] 1 Microsphere coating: The polystyrene microspheres are coated by physical adsorption method, and the Fab segment remains active by utilizing the hydrophobic effect of the Fc segment of the antibody to adsorb on the microspheres. Resuspend the microspheres before coating, add the microspheres to the mouse anti-human platelet membrane glycoprotein monoclonal antibody diluted in the coating buffer (make the ratio of antibody to microspheres 70ug antibody / ml microspheres), and vortex at room temperature for 2 Hours (or overnight at 4°C with vortex shaking), washed three times with 0.01M PBS / TWEEN, then blocked the microspheres with 5% bovine serum albumin shaking for 2 hours, washed three times with 0.01M PBS / TWEEN, and washed in an isotonic medium containing 0.2% sodium azide Resuspend the microspheres in saline solution to a final concentration of 0.25% microspheres. Store at 4°C. On days 1, 2, 5, 10, 30...

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Abstract

The invention provides a method belonging to the field of cytometric bead array with high sensitivity for testing platelet specific antibody based on platelet basic experimental study. Platelet lysate is used to incubate the microsphere which is coated with platelet membrane glycoprotein monoclonal antibody, then goat-anti-human IgG polyclonal antibody labeled with phycoerythrin is added, which is analyzed in cytometric bead array. If autoantibodies exist on the surface of platelet, 'microsphere-platelet membrane glycoprotein monoclonal antibody-platelet membrane glycoprotein autoantibodies-goat-anti-human IgG polyclonal antibody labeled with phycoerythrin' complex structure is formed, the fluorescence intensity of the test microsphere is enhanced. The invention is easy to operate, and has mature technology, high sensitivity for testing platelet autoantibodies, which is good for basic experimental study of platelet antibody.

Description

Technical field: [0001] The invention belongs to a novel experimental method with high sensitivity for detecting platelet-specific antibodies in the basic experimental research of platelets. Background technique: [0002] According to the time when the platelet antibody detection method appears in the laboratory, it can be divided into phase I, phase II and phase III examination methods. Phase I examination appeared in the 1850s to 1970s, mainly to detect the effect of patient's serum or plasma on normal platelet function in vitro. The experimental methods used are all indirect methods with low sensitivity and specificity. Phase II assays were developed in the 1970s to detect platelet-associated antibodies. Platelet-associated antibodies have high sensitivity but low specificity. The Phase III experimental method is an antigen-specific assay developed in the mid-1980s. The recently developed glycoprotein capture assay—the monoclonal antibody immobilized specific platelet ...

Claims

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Application Information

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IPC IPC(8): G01N33/577G01N33/546
Inventor 侯明冀学斌彭军刘新光张爱军石艳秦平
Owner 侯明
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