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A reagent card for detecting the curative effect of thienopyridine antiplatelet drugs and its use method and application

A technology of anti-platelet drugs and detection reagents, which is applied in the direction of testing pharmaceutical preparations, biological tests, material inspection products, etc., to achieve practical effects.

Active Publication Date: 2018-01-26
北京乐普诊断科技股份有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] However, there is currently no report on the use of optical turbidimetry to detect platelet aggregation in whole blood samples. Therefore, there is an urgent need in this field to develop a detection reagent that can more accurately detect platelet aggregation.

Method used

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  • A reagent card for detecting the curative effect of thienopyridine antiplatelet drugs and its use method and application
  • A reagent card for detecting the curative effect of thienopyridine antiplatelet drugs and its use method and application
  • A reagent card for detecting the curative effect of thienopyridine antiplatelet drugs and its use method and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0060] microsphere labeling

[0061] (1) Washing balls: take 100 μL of fluorescent microspheres, wash twice with 1 mL of MES pH4.5 buffer, centrifuge at 8000 g, 4°C for 10 min, discard the supernatant, and reconstitute to 1 mL by adding MES pH4.5 after ultrasonication for one minute.

[0062] (2) Activation of microspheres: Add EDC and NHS solutions to a final concentration of 0.5mg / mL, activate at room temperature for 15 minutes, centrifuge at 4°C (8000g, 10min), discard the supernatant, and use 0.02M pH8.5 borate buffer 1 mL was washed twice and reconstituted to 1 mL with this buffer.

[0063] (3) Add labeled protein: Sonicate the activated microspheres for one minute, mix with a vortex mixer, add GpIIb / IIIa receptor ligand to a final concentration of 0.5 mg / mL, and incubate at room temperature for 2 hours.

[0064] (4) Blocking: after the incubation, BSA with a final concentration of 0.5% was added and blocked at room temperature for 30 min.

[0065] (5) Centrifugation: A...

Embodiment 2

[0073] 1. Inhibition of GpIIb / IIIa receptor activity

[0074] Blood samples were collected from 31 healthy volunteers, anticoagulated with 3.2% sodium citrate, and 2 tubes were taken from each case, each 2 mL. An appropriate amount of GpIIb / IIIa receptor activity inhibitor abciximab (Reopro) was dissolved in 0.01M PBS buffer to prepare 0.15mM. One blood sample was taken from each case as the experimental group, 2 μL of 0.15 mM abciximab was added respectively, and the final concentration of abciximab in the blood sample was 0.15 μM. Gently invert the blood sample 10 times to mix the abciximab and the blood sample thoroughly, and incubate at 37 degrees for 1 hour. 2 μL of 0.01M PBS buffer solution was added to each remaining blood sample, and incubated at 37°C for 1 hour, which was used as a control group.

[0075] GpIIb / IIIa receptor activity assay

[0076] After the above-mentioned experimental incubation, take the blood samples of the experimental group and detect the GpI...

Embodiment 3

[0078] Clopidogrel Drug Inhibition Rate Evaluation Test

[0079]Collect 2 mL of sodium citrate whole blood from patients taking clopidogrel, and gently invert the blood collection tube several times to mix well. Insert the thienopyridine antiplatelet drug curative effect detection reagent card into the matching optical turbidimetric detection equipment, insert the blood collection tube into the sampling needle of the reagent card, and click Start to start the detection. The instrument automatically detects the change degree of the light transmittance of the four detection slots of the reagent card, and converts it into an electrical signal through the corresponding software, draws the change curve of the electrical signal, and finally gives the platelet aggregation rate and drug inhibition rate. For platelet aggregation detection results of the four channels of the reagent card, see image 3 . The platelet aggregation percentage PAP% of the ADP activation pathway measured in...

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Abstract

The invention provides a reagent card for detecting the therapeutic effect of thienopyridine antiplatelet drugs and an application method and application of the reagent card. The reagent card comprises four detection channels which are sequentially arranged in parallel and are independent of one another, the detection channel I contains channel I detection reagent, and the channel I detection reagent contains a lyophilized agent, wherein the lyophilized agent is obtained by firmly bonding activated dye microspheres with receptor ligand of blood platelets GpIIb / IIIa through amido bonds, then adding blood coagulation factors, and conducting freeze-drying; the detection channel II contains channel II detection reagent, and the channel II detection reagent contains a lyophilized agent serving as a blood platelet maximum activating agent; the detection channel III contains no detection reagent, the detection channel IV contains channel IV detection reagent, and the channel IV detection reagent contains a lyophilized agent serving as a blood platelet P2Y12 receptor activating agent and P2Y1 receptor antagonist. By means of the reagent card, the therapeutic effect of thienopyridine antiplatelet drugs can be detected easily, fast, conveniently and accurately.

Description

technical field [0001] The invention belongs to the field of drug curative effect detection reagents, and relates to a reagent card for detecting the curative effect of thienopyridine antiplatelet drugs and its use method and application. Background technique [0002] Clinical studies have shown that vascular lesions are the fundamental pathological basis of cardiovascular and cerebrovascular diseases. The decrease in arterial vascular elasticity will lead to increased blood pressure during systole and decreased blood pressure during diastole, which will increase the pulse pressure difference; if coronary artery atherosclerosis Coronary heart disease can result from the formation of plaques or blood clots. The thrombus formed in the coronary artery can be divided into three types according to the color and shape, namely, white thrombus, red thrombus and mixed red and white thrombus. White thrombus is mainly formed by abnormal adhesion and aggregation of platelets. This kind...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/15G01N33/53
CPCG01N33/15G01N33/53
Inventor 陈勇田宇王玲
Owner 北京乐普诊断科技股份有限公司
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