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Kit for quantitatively detecting D-dimer and preparation method thereof

A quantitative detection and dimer technology, applied in the field of biomedicine, can solve the problems of poor specificity and low sensitivity, and achieve the effect of high specificity, high sensitivity and high accuracy

Inactive Publication Date: 2018-08-24
浙江艾明德生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The invention provides a kit and preparation method for quantitative detection of D-dimer, which overcomes the shortcomings of low sensitivity and poor specificity in the prior art

Method used

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  • Kit for quantitatively detecting D-dimer and preparation method thereof
  • Kit for quantitatively detecting D-dimer and preparation method thereof
  • Kit for quantitatively detecting D-dimer and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0051] Example 1 Preparation of D-dimer acridinium lipid-labeled monoclonal antibody

[0052] 1) Weigh an appropriate amount of D-dimer monoclonal antibody, and use 0.05mol / L pH 9.5 CB to adjust to 1mg / mL;

[0053] 2) Acridine lipid and antibody are calculated according to the molar ratio of 1:15, dissolved in DMF, mixed, and reacted at room temperature for 30 minutes;

[0054] 3) The reaction solution was transferred to a dialysis bag (molecular weight cut-off 8000-12000), and dialyzed with 0.01M PBS pH 7.2 for 24 hours;

[0055] 4) Add an equal volume of glycerin and 0.1% PC-300.

Embodiment 2

[0056] Example 2 Preparation of the D-dimer quantitative assay kit of the present invention

[0057] 1. Preparation of acridine lipid antibody

[0058] 1. Acridine-labeled monoclonal antibody diluent Buffer A:

[0059] Buffer A 0.01-0.1M Tris-HCl pH 7.6, mass ratio 0.1%-1% deb bovine serum albumin (BSA), volume ratio 0.2% PC-300;

[0060] 2. Select the concentration of acridine-labeled monoclonal antibody:

[0061] The working concentration of acridinium-labeled monoclonal antibody is greater than 0.5 μg / mL.

[0062] 2. Preparation of D-dimer calibrator

[0063] Dilute the D-dimer antigen with 0.01M PBS pH 7.2, BSA diluent with a mass ratio of 0.5%, and prepare five kinds of calibration products with standard concentrations of 0.2, 1, 5, 20, and 50 μg / mL.

[0064] 3. Preparation of antibodies labeled with acridine derivatives:

[0065] 1) Weigh an appropriate amount of D-dimer monoclonal antibody, and use 0.05mol / L pH 9.5 CB to adjust to 1mg / mL;

[0066] 2) Acridine lipi...

Embodiment 3

[0091] Example 3 Buffer Buffer

[0092] As an important component of the present invention, the buffer is a buffer for diluting biotinylated antibodies, antibodies labeled with acridine derivatives, and streptavidin-coated magnetic particles, so as to further reduce specific binding and improve the analytical sensitivity of reagents.

[0093] Buffer A: 0.01-0.1M Tris-HCl pH 7.6, mass ratio 0.1%-1%BSA, volume ratio 0.2% PC-300;

[0094] Buffer B: 0.01-0.1M 4-hydroxyethylpiperazineethanesulfonic acid (HEPES) at pH 7.8, 5% mouse serum by volume, 0.5-2mg / mL sheep immunoglobulin G, 0.1%-5 by mass % trehalose, 0.5%-1% casein sodium salt by mass ratio, and 0.2% PC-300 by volume ratio.

[0095] In one of the embodiments, trehalose can also be sucrose or fructose, and some experimental data are as follows:

[0096]

[0097] Analytical Sensitivity=2*(0 value RLU SD)*5 / (Calibrator RLU mean - 0 value RLU mean)

[0098] Use Buffer B as the diluent of acridinium ester-D-dimer antibody...

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Abstract

The invention provides a kit for quantitatively detecting D-dimer. The kit includes: a D-dimer calibrator, magnetic-particles coated with streptavidin, a D-dimer monoclonal antibody marked with an acridine derivative, a D-dimer monoclonal antibody marked with a biotin, a chemical illumination substrate, and a quality control substance. The preparation method of the kit includes the following steps: preparing the calibrator from a D-dimer pure product raw material; preparing the antibody marked with the acridine derivative, preparing the biotinylated antibody; coating magnetic-particles with the streptavidin; dispensing the calibrator, the marker mixture liquid and the chemical illumination substrate; and combining the finish product. The kit has high sensitivity and good specificity, is high in accuracy of a quantitative determination result, is low in use cost and is easier to promote and apply.

Description

technical field [0001] The invention relates to the field of biomedicine, in particular to a kit for quantitatively detecting D-dimer and a preparation method. Background technique [0002] In the process of hemostasis and thrombus, thrombin hydrolyzes plasma fibrinogen to form fibrinogen monomers, which are linked into fibrin; at the same time, thrombin activates factor VIII to cross-link fibrinogen; finally, under the action of plasmin, Fibrin is degraded into various fibrin degradation products (FDP), the smallest fragment of which is D-dimer (D-dimer), the molecular weight is about 62KD, and the half-life in the body is about 3h, mainly through renal excretion and reticuloendothelial system destruction . [0003] The generation of D-dimer reflects the activation of the coagulation and fibrinolytic system of the body, and it is related to the increase of D-dimer in various thrombotic diseases and physiological hyperagglutination states. D-dimer and disseminated vascular...

Claims

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Application Information

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IPC IPC(8): G01N33/577G01N21/76
CPCG01N21/76G01N33/577G01N2800/32
Inventor 陈志强胡国富朱炎
Owner 浙江艾明德生物科技有限公司
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