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Thromboelastometry heparin quantitative detection kit and preparation method thereof

A thromboelastography and quantitative detection technology, applied in the field of medical devices, can solve the problems of inability to detect whole blood, quantitative detection of heparin, and inapplicability of bedside detection

Active Publication Date: 2021-05-11
上海原科实业发展有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0026] To sum up, in the prior art: when using APTT or PT detection, only when all the proteins in the whole coagulation cascade are intact can the heparin level be accurately measured, but reagents and instruments will affect its sensitivity to heparin, Causes up to four-fold variability in final results between laboratories; protamine sulfate neutralization test only for unfractionated heparin (UFH) monitoring; coagulation activation assay kit (coagulation method) and heparinase-coated reagents The cup cannot quantitatively detect heparin; in addition, APTT or PT and protamine sulfate neutralization test and anti-Xa activity detection and one-step detection are only suitable for detecting the amount of heparin in plasma. When testing, patient samples need to be processed to obtain plasma It cannot be tested directly on whole blood, so it is not suitable for bedside testing

Method used

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  • Thromboelastometry heparin quantitative detection kit and preparation method thereof
  • Thromboelastometry heparin quantitative detection kit and preparation method thereof
  • Thromboelastometry heparin quantitative detection kit and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] S11) Weigh 0.596g of buffering agent 4-hydroxyethylpiperazineethanesulfonic acid (English abbreviation: HEPES) and put it into a 250ml volumetric flask, dissolve it with an appropriate amount of distilled water, and then add an appropriate amount of NaOH solution with a concentration of 0.1mol / L. Adjust the pH value, add water to the mark, mix well, and prepare a HEPES buffer solution with a concentration of 10mmol / L and a pH value of about 7.9;

[0046] S12) Draw distilled water to redissolve the lyophilized powder of coagulation factor Xa to make a stock solution of coagulation factor Xa with a concentration of 1 g / L;

[0047] S13) Draw distilled water to redissolve the lyophilized powder of the coagulation factor activator RVV-V to make a stock solution of the coagulation factor activator RVV-V with a concentration of 1 g / L;

[0048] S14) Weigh the rabbit brain curd fat and add the HEPES buffer with the weight of the rabbit brain curd fat: volume = 1:1, grind until m...

Embodiment 2

[0054] S21) Weigh 1.489g of buffering agent 4-hydroxyethylpiperazineethanesulfonic acid and put it into a 250ml volumetric flask, dissolve it with an appropriate amount of distilled water, add an appropriate amount of NaOH solution with a concentration of 0.1mol / L to adjust the pH value, and then add water to the mark, mix evenly, and prepare HEPES buffer solution with a concentration of 25mmol / L and a pH value of about 7.4;

[0055] S22), S23), S24) are respectively identical with embodiment 1 step S12), S13), S14);

[0056] S25) Weigh 0.877g of NaCl, 0.4g of glycine, 2.0g of bovine serum albumin, and 2.5g of trehalose, add them to a 100ml volumetric flask, and add an appropriate amount of HEPES buffer to prepare a support solution;

[0057] S26) Draw 2.0ml coagulation factor Xa stock solution, 0.6ml blood coagulation factor activator stock solution, 0.5ml rabbit brain fat coagulation stock solution and add to the volumetric flask equipped with support solution;

[0058] S27...

Embodiment 3

[0061] S31) Weigh 2.979g of buffering agent 4-hydroxyethylpiperazineethanesulfonic acid (English abbreviation: HEPES) into a 250ml volumetric flask, add appropriate amount of distilled water to dissolve, then add an appropriate amount of NaOH solution with a concentration of 0.1mol / L Adjust the pH value, add water to the scale line, mix well, and prepare a HEPES buffer solution with a concentration of 50mmol / L and a pH value of about 7.1;

[0062] S32), S33), S34) are respectively identical with embodiment 1 step S12), S13), S14);

[0063] S35) Weigh 1.695g of NaCl, 0.6g of glycine, 3.0g of bovine serum albumin, and 3.8g of trehalose, add them to a 100ml volumetric flask, and add an appropriate amount of HEPES buffer to prepare a support solution;

[0064] S36) Draw 3.0ml coagulation factor Xa stock solution, 1.0ml blood coagulation factor activator stock solution, 1.0ml rabbit brain fat coagulation stock solution and add them into the volumetric flask equipped with support so...

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Abstract

A kit for quantitative detection of heparin by thromboelastography, comprising a buffer, blood coagulation factor Xa, rabbit brain fat coagulation, blood coagulation factor activator, support agent, biological preservative; the preparation method includes: preparing buffer solution, blood coagulation factor Xa stock solution, Blood coagulation factor activator stock solution, biological antiseptic solution, rabbit brain lipid coagulation stock solution; take support agent and add buffer solution to prepare support solution; add coagulation factor Xa, coagulation factor activator, rabbit brain fat coagulation stock solution according to kit specifications In the support solution, add biological preservative solution after mixing, and then add buffer solution to the specified amount, mix and freeze-dry. The invention can quantitatively detect the heparin in the sample by the thromboelastography detection method, the correlation is good in the linear range, meets the detection limit standard of heparin, and the coefficient of variation is not more than 10%. There is also no requirement for the integrity of the protein in the waterfall, which simplifies the inspection process and is convenient for doctors and patients.

Description

technical field [0001] The invention relates to a thromboelastography heparin quantitative detection kit and a preparation method thereof, belonging to the technical field of medical devices. Background technique [0002] Heparin, named after it was first discovered in the liver, also exists in the lungs, blood vessel walls, intestinal mucosa and other tissues, and is an alternate form of glucosamine, L-iduroside, N-acetylglucosamine and D-glucuronic acid The mucopolysaccharide sulfate composed of it is strongly acidic and is a natural anticoagulant substance in animals. It naturally exists in mast cells and is mainly extracted from bovine lung or pig small intestinal mucosa. [0003] The heparin initially used clinically is called standard heparin, unfractionated heparin or unfractionated heparin (Unfractionated Heparin), with an average molecular weight of 15KD. Antithrombotic, and anticoagulant treatment in hemodialysis, extracorporeal circulation, catheterization, micro...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/66
CPCG01N33/66G01N2400/40
Inventor 王连升李红梅
Owner 上海原科实业发展有限公司
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