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Determining method of thrombin inhibition activity

A technology of thrombin inhibitor and inhibitory activity, which is applied in the field of anticoagulation, can solve the problems of narrow application range and inability to consider the external site of thrombin, etc., achieve wide application range, optimize components and detection methods, and improve The effect of accuracy

Pending Publication Date: 2019-11-08
DALIAN POLYTECHNIC UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The purpose of the present invention is to solve the problem that the existing thrombin inhibitor detection method has a narrow adaptability range and cannot take into account the external position of thrombin, so as to provide a new method for measuring thrombin inhibitory activity with a wider application range

Method used

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  • Determining method of thrombin inhibition activity
  • Determining method of thrombin inhibition activity
  • Determining method of thrombin inhibition activity

Examples

Experimental program
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Effect test

specific Embodiment approach 1

[0034] Specific implementation mode 1: The detection method of thrombin inhibitor activity of this embodiment is implemented in the following steps:

[0035] 1. Configure buffer solution, configure imidazole concentration to be 0.05mol / L, sodium chloride concentration to be 0.1mol / L, calcium chloride concentration to be 0.002mol / L or Tris concentration to be 0.05mol / L, sodium chloride concentration to be 0.1mol / L, a buffer with a calcium chloride concentration of 0.002mol / L. Adjust the buffer pH to 7.4±0.05 with hydrochloric acid;

[0036] 2. Dissolve the inhibitor to be tested, fibrinogen, and thrombin with imidazole buffer. Place the fibrinogen solution at 37°C and the thrombin solution at 4°C for use;

[0037] 3. Add 200μL of fibrinogen solution and 100μL of inhibitor or buffer solution to the blood coagulation cup, incubate at 37°C, and then add 100μL of thrombin solution to the blood coagulation cup to detect the time required for fibrinogen to coagulate;

[0038] 4. Determine...

specific Embodiment approach 2

[0039] Specific embodiment two: This embodiment is different from the specific embodiment one in that the concentration of the fibrinogen solution in step two is 1 to 1.5 mg / mL. Other steps and parameters are the same as in the first embodiment.

specific Embodiment approach 3

[0040] Specific embodiment three: This embodiment is different from specific embodiment one or two in that the concentration of the thrombin solution in step two is 8-12 U / mL. Other steps and parameters are the same as those in the first or second embodiment.

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Abstract

The invention discloses a determining method of thrombin inhibition activity. A fibrinogen solution and a to-be-tested sample solution are mixed, and the mixture is added to a blood blotting cup containing magnetic beads to serve as a detection group; a buffer solution of the same volume is used to replace the to-be-tested sample solution in the detection group to serve as a blank group; the detection group and the blank group are placed at 37 DEG C for 60-300 s, the thrombin solution in the S1 is added, the setting time of the solution in the blood blotting cup is detected by a full-automaticblood clotting analyzer, significance analysis is performed on the setting time by t test, and if the setting time of the detection group is significantly longer than the blank group (P is smaller than 0.05), the to-be-tested sample is degraded to have thrombin inhibition activity. Components of the buffer solution and the detection means are optimized and improved, and false positive results during detection by existing methods are effectively avoided; and the range of detachable samples is enlarged and larger and more accurate sample space is provided for detection.

Description

Technical field [0001] The invention relates to the technical field of anticoagulation, in particular to a method for measuring thrombin inhibitory activity. Background technique [0002] Due to the high work pressure and fast pace of life of modern people, as well as the influence of poor living habits such as insufficient sleep, excessive eating, smoking and alcohol abuse, thrombotic diseases in cardiovascular and cerebrovascular diseases have become the number one killer of mankind. At present, cardiovascular diseases cause millions of premature deaths in Europe every year, and the prevalence of cardiovascular diseases in my country is on a continuous rise. It is estimated that there are 290 million patients suffering from cardiovascular diseases, and the number of patients with cardiovascular diseases will continue to grow rapidly in the next 10 years. According to investigations, cardiovascular deaths account for the first cause of death among urban and rural residents, 45....

Claims

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Application Information

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IPC IPC(8): C12Q1/56
CPCC12Q1/56G01N2333/974
Inventor 杜明刘汉雄涂茂林程述震孙黎明王震宇袁禛乔昕昱
Owner DALIAN POLYTECHNIC UNIVERSITY
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