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A reagent for the determination of antithrombin III

A technology of antithrombin and bovine thrombin, which is applied in the direction of measuring devices, material analysis and instruments by observing the influence on chemical indicators, and can solve the problems of high price

Active Publication Date: 2021-10-26
山东艾科达生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At the same time, it can maintain the activity of enzymes in the system, but because of its high price, it is only used in clinical research in a small amount

Method used

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  • A reagent for the determination of antithrombin III
  • A reagent for the determination of antithrombin III
  • A reagent for the determination of antithrombin III

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Preparation of Reagent 1 and Reagent 2

[0032] Preservative 1: 0.5 mg / mL sodium azide.

[0033] Preservative 2: 0.02% Thimerosal.

[0034] Preservative 3: 0.02% ProClin300.

[0035] Preservative 4: 0.3w% pomelo peel extract, 0.5w% rhubarb extract, 0.5w% allicin.

[0036] Preparation of Reagent 1:

[0037] The reagent 1 is put into the component raw materials in order, and the next one is added after ensuring that each component raw material is completely dissolved. Weigh 0.24228g of Tris biological buffer, add 90mL of purified water, and adjust the pH to pH7.4; add inorganic salts 0.73g; bovine serum albumin 2g; heparin 500IU; bovine thrombin 500IU, add preservatives, mix slowly, and dilute to 100mL with purified water;

[0038] For the reagent 2, add the component raw materials in order to ensure that each component raw material is completely dissolved before adding the next one, weigh 0.24228g of Tris biological buffer, add 90mL of purified water, adjust the pH t...

Embodiment 2

[0046] The analytical performance evaluation of embodiment 2 kit of the present invention

[0047] (1) Analytical sensitivity

[0048] Prepare reagent 1 and reagent 2 according to the method of example 1, choose preservative 4. The prepared antithrombin III (chromogenic substrate method) kit tested samples with a concentration of 90-100%, repeated the test 3 times, calculated the average value of the dOD values ​​obtained from the 3 tests, and the results were all greater than 0.600. as shown in Table 3:

[0049] Table 3 Analytical Sensitivity Determination Results

[0050]

[0051] (2) Linear range

[0052] Dilute into 6 dilution concentrations with the high concentration sample (AT-III activity is 145%) close to the upper limit of the linear range of antithrombin III and the diluent, as shown in the table below; each dilution concentration is tested 3 times, and the results of the determination are obtained respectively. mean. Take the dilution concentration as the i...

Embodiment 3

[0061] Embodiment 3 stability investigation

[0062] The reagent prepared by the method disclosed in CN107153043A embodiment 1 is used as contrast reagent, (reagent: 0.2U / mLFXa, 0.3mol / L tris-hydrochloric acid buffer solution pH6.5, 1%mol / L sodium chloride and 2% BSA, 2% PEG6000, 10% mannitol, 0.05% Proclin300. Reagent: redissolve the chromogenic substrate N-α-Z-D-Arg-Gly-Arg-pNA·2HCl, adjust the pH to 6.5 with acetic acid, add Mannitol and sodium azide are adjusted to a concentration of 1.5mmol / L with water, the concentration of mannitol is 5%, and the concentration of sodium azide is 0.05%.) and the reagents of the present invention are respectively placed at 2-8°C and 37°C, each Samples were taken every 7 days, and the quality controls (target value range 92-112%, target value 102%) were determined respectively. The test results are shown in Table 4

[0063] Table 4

[0064]

[0065]

[0066] The test results show that the liquid ready-to-use antithrombin III activ...

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Abstract

The invention provides a reagent for measuring antithrombin III, comprising reagent 1 and reagent 2; said reagent 1 comprises bovine thrombin with a concentration of 2-10IU / mL, heparin with a concentration of 1-10IU / mL, Tris biological buffer solution with a concentration of 6.3g / L-8.3g / L and bovine serum albumin, sodium chloride, and preservatives at a concentration of 5w%; the reagent 2 includes a chromogenic substrate with a concentration of 2-4w%, Tris biological buffer, 5w% mannitol, preservatives, 0.3w% pomelo peel extract, 0.5w% rhubarb extract, 0.5w% allicin preservative synergistic effect, can play a better sterilization Antibacterial effect, and natural biological preservatives are non-toxic, non-polluting, non-corrosive, have no potential danger to users and the environment, and can play the role of protective agent to improve the performance index of the kit. Regardless of whether it is stored at 37°C or 2-8°C, its bactericidal and antibacterial properties are good, and the reagents have high stability. Therefore, the present invention has high clinical application value and potential market benefit.

Description

technical field [0001] The invention belongs to the technical field of biomedical detection, and in particular relates to a reagent for measuring antithrombin III containing a compound preservative. Background technique [0002] Antithrombin III (AT-III) is a single-chain glycoprotein synthesized by liver and vascular endothelial cells, which can combine with heparin and thrombin to exert its anticoagulant effect. Antithrombin Ⅲ (AT-Ⅲ) mainly reflects the function of the body's anticoagulant system, and is a good index for clinically evaluating the hypercoagulable state. [0003] There are mainly three methods for clinically measuring AT-Ⅲ activity: coagulation method, immunoassay and chromogenic substrate method. In the present invention, the chromogenic substrate method is used. Compared with other methods, the method has high sensitivity, good accuracy, and short detection time, and can be applied to various automatic analysis instruments, greatly reducing the cost of cl...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N21/78G01N33/573
CPCG01N21/78G01N33/573
Inventor 谯兴强许玉荣王醒谭柏清赵志敏
Owner 山东艾科达生物科技有限公司
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