Reagent card for detecting therapeutic effect of thienopyridine antiplatelet drugs and application method and application of reagent card

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

Active Publication Date: 2016-06-29
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.

Method used

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  • Reagent card for detecting therapeutic effect of thienopyridine antiplatelet drugs and application method and application of reagent card
  • Reagent card for detecting therapeutic effect of thienopyridine antiplatelet drugs and application method and application of reagent card
  • Reagent card for detecting therapeutic effect of thienopyridine antiplatelet drugs and application method and application of reagent card

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0060] Microsphere marking

[0061] (1) Wash the balls: Take 100μL of fluorescent microspheres, wash them twice with 1mL of MESpH4.5 buffer solution, 8000g, centrifuge for 10min at 4°C, discard the supernatant, add MESpH4.5 to 1mL after sonicating for one minute.

[0062] (2) Activated 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) to discard the supernatant, and use 1mL of 0.02MpH8.5 borate buffer After washing twice, use the buffer to reconstitute to 1 mL.

[0063] (3) Adding the labeled protein: ultrasound the activated microspheres for one minute, vortex the mixer to mix, add the 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, add BSA with a final concentration of 0.5% and block for 30 minutes at room temperature.

[0065] (5) Centrifugation: After closing, centrifuge at...

Embodiment 2

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

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

[0075] GpIIb / IIIa receptor activity detection

[0076] After the above-mentioned experimental incubation, take the blood sample of the experimental group to detect the ...

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 for several times. Insert the thienopyridine antiplatelet drug efficacy test reagent card into the matching optical turbidity test equipment, insert the blood collection tube into the injection needle of the reagent card, click Start to start the test. The instrument automatically detects the light transmittance change degree of the four detection slots of the reagent card, and converts it into electrical signals through the corresponding software, draws the electrical signal change curve, and finally gives the platelet aggregation rate and drug inhibition rate. The platelet aggregation test results of the four channels of the reagent card are shown in image 3 . The platelet aggregation percentage PAP% of the ADP activation pathway measured in this example is 29.7%, and the corresponding clopido...

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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 efficacy detection reagents, and relates to a reagent card for detecting the efficacy of thienopyridine antiplatelet drugs, and a use method and application thereof. Background technique [0002] Clinical studies have shown that angiogenesis is the fundamental pathological basis for cardiovascular and cerebrovascular diseases. The decrease in arterial vascular elasticity will lead to an increase in blood pressure during vasoconstriction, a decrease in diastolic blood pressure, and an increase in pulse pressure; if coronary artery atherosclerosis occurs Plaque-like plaques or thrombosis can lead to coronary heart disease. The thrombus formed by coronary arteries can be divided into three types in terms of 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 of thrombus mostly occurs in the...

Claims

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Application Information

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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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