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Antithrombin III activity determination kit and preparation method thereof

An activity measurement and antithrombin technology, applied in the field of clinical testing, can solve the problems of interfering with antithrombin III activity detection, increasing the production cost of the kit, easy inactivation of activation factor X, etc., and achieving the consistency of the difference between bottles Good, improve accuracy, avoid the effect of reaction rate

Active Publication Date: 2021-04-16
BEIJING MDC NEW SPRING MEDICAL DEVICES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, activating factor X (FXa) is easily inactivated and expensive, which increases the cost of kit production
[0007] Both thrombin-based and activated factor X (FXa)-based substrate chromogenic methods require the presence of excess thrombin or FXa. According to the coagulation cascade theory, other coagulation factors in plasma are more or less Both can consume or inactivate the activity of thrombin and activated factor X, thereby interfering with the detection of antithrombin III activity
[0008] The current chromogenic substrate method for detection of antithrombin III activity still needs to be improved

Method used

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  • Antithrombin III activity determination kit and preparation method thereof
  • Antithrombin III activity determination kit and preparation method thereof
  • Antithrombin III activity determination kit and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] Preparation of dilution buffer: Weigh 0.075g of benzalkonium bromide and dissolve in 100mL of normal saline.

[0043] Preparation of thrombin reagent: the distribution ratio of each component is 4.5U / mL bovine thrombin, 3.5U / mL heparin sodium, 12% (w / v) trehalose, 0.25% (w / v) lysine hydrochloride , 2% (w / v) Sodium Chloride, 0.4% (v / v) Protein Stabilizer II, 1% (v / v) 60mmol / L, pH8.0 tris-hydrochloric acid buffer.

[0044] Preparation of substrate reagent: prepare 75mmol / L Tris-HCl buffer containing 0.1% (w / v) thimerosal sodium, 5% (w / v) lactose, 7% (w / v) trehalose, pH8.0; Finally, add thrombin substrate PA2493, stir and mix until the concentration is 1.2mmol / L.

[0045] After the above-mentioned thrombin reagent and substrate reagent are prepared, they are subpackaged and freeze-dried to make freeze-dried powder.

Embodiment 2

[0047] Preparation of dilution buffer: 100 mL of physiological saline.

[0048] Preparation of thrombin reagent: the distribution ratio of each component is 4.5U / mL bovine thrombin, 3.5U / mL heparin sodium, 12% (w / v) trehalose, 0.25% (w / v) lysine hydrochloride , 2% (w / v) Sodium Chloride, 0.4% (v / v) Protein Stabilizer II, 1% (v / v) 60mmol / L, pH8.0 tris-hydrochloric acid buffer.

[0049] Preparation of substrate reagent: prepare 75mmol / L Tris-HCl buffer containing 0.1% (w / v) thimerosal sodium, 5% (w / v) lactose, 7% (w / v) trehalose, pH8.0; Finally, add thrombin substrate PA2493, stir and mix until the concentration is 1.2mmol / L.

[0050] After the above-mentioned thrombin reagent and substrate reagent are prepared, they are subpackaged and freeze-dried to make freeze-dried powder.

Embodiment 3

[0052] Preparation of dilution buffer: Weigh 0.075g of benzalkonium bromide and dissolve in 100mL of normal saline.

[0053] Preparation of thrombin reagent: the distribution ratio of each component is 4.5U / mL bovine thrombin, 3.5U / mL heparin sodium, 12% (w / v) trehalose, 0.25% (w / v) sodium chloride, 60mmol / L, pH8.0 tris-hydrochloric acid buffer.

[0054] Preparation of substrate reagent: prepare 75mmol / L Tris-HCl buffer containing 0.1% (w / v) thimerosal sodium, 5% (w / v) lactose, 7% (w / v) trehalose, pH8.0; Finally, add thrombin substrate PA2493, stir and mix until the concentration is 1.2mmol / L.

[0055] After the above-mentioned thrombin reagent and substrate reagent are prepared, they are subpackaged and freeze-dried to make freeze-dried powder.

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Abstract

The invention relates to the field of clinical examination, in particular to an antithrombin III activity determination kit and a preparation method thereof. The kit is prepared from the following raw materials: a dilution buffer solution, a thrombin reagent and a chromogenic substrate reagent, The dilution buffer solution is normal saline containing benzalkonium bromide; The thrombin reagent comprises thrombin, heparin sodium, trehalose, sodium chloride and a buffer solution; and the chromophoric substrate reagent comprises a chromophoric substrate, lactose, trehalose and a buffer solution. Benzalkonium bromide is added into the dilution buffer solution, so that the structure or state of fibrinogen in blood plasma can be changed, the fibrinogen is prevented from interfering with the reaction rate of thrombin splitting substrates, and the accuracy of detecting the activity of antithrombin III is improved.

Description

technical field [0001] The invention relates to the field of clinical testing, in particular to an antithrombin III activity assay kit and a preparation method thereof. Background technique [0002] The promotion and inhibition of the blood coagulation process in the normal human body are in a state of dynamic balance. As the most important anticoagulant substance in the human body, antithrombin III (hereinafter referred to as AT-III) controls the process of blood coagulation and fibrinolysis, thereby maintaining the coagulation balance of the body, so the change of AT-III activity is a diagnostic diffuse It is an important indicator of chronic intravascular coagulation (DIC), liver cirrhosis, sepsis, thrombotic diseases (myocardial infarction, venous thrombosis, etc.), congenital AT-Ⅲ deficiency, hemophilia, and aplastic anemia. [0003] AT-III is a globulin that is synthesized in the liver. Studies have shown that low levels or activity of antithrombin III can be either ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/56
Inventor 邹红建胡军隋延锋
Owner BEIJING MDC NEW SPRING MEDICAL DEVICES
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