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A kind of antithrombin Ⅲ assay kit and its detection method

A technology of antithrombin and reagent kits, applied in the field of biomedical testing, can solve problems such as large application limitations, complicated operation, and poor stability, and achieve the effects of strong anti-interference ability, wide application prospects, and good stability

Active Publication Date: 2020-06-19
NINGBO RUI BIO TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] However, the chromogenic substrate method also has certain defects. This method is to analyze the AT-III activity in the sample by adding exogenous thrombin, because there is also another heparin-catalyzed thrombin inhibitor—heparin cofactor II in the sample. (HCII), so the activity of determining the remaining thrombin is the remaining activity of thrombin after reacting with AT-III and HCII, therefore, this method is susceptible to the interference of HCII in the sample, resulting in inaccurate determination of AT-III
[0006] Currently commercially available reagents mainly use heparinase to degrade heparin to reduce the interference of HCII, but the operation is complicated and the heparinase is expensive, which greatly increases the cost
The other is through the coagulation factor Xa pathway. Although it avoids the interference of heparin cofactor II, the coagulation factor Xa is not only expensive, but also has poor stability. The sensitivity of the reagent is not as high as that of the thrombin pathway, and its application is very limited.

Method used

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  • A kind of antithrombin Ⅲ assay kit and its detection method
  • A kind of antithrombin Ⅲ assay kit and its detection method
  • A kind of antithrombin Ⅲ assay kit and its detection method

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Embodiment 1

[0039] The antithrombin III assay kit of this embodiment consists of the following components:

[0040] Reagent R1

[0041] Tris buffer 100mmol / L, BSA 5g / L, gelatin 50g / L, mannitol 50g / L, ammonium sulfate 1mol / L, Triton-100 1ml / L, sodium azide 1g / L, trehalose 50g / L , heparin 10mg / L, thrombin 10ku / L.

[0042] Reagent R2

[0043] Tris buffer 100mmol / L, BSA 5g / L, PVP 10g / L, trehalose 50g / L, Tween-20 1ml / L, sodium azide 1g / L, HD-Phe-Pip-Arg-pNA·2HCl 2g / L.

Embodiment 2

[0045]The antithrombin III assay kit of this embodiment consists of the following components:

[0046] Reagent R1

[0047] Tris buffer 100mmol / L, BSA 5g / L, mannitol 50g / L, ammonium sulfate 2mol / L, Triton-100 1ml / L, sodium azide 1g / L, trehalose 50g / L, heparin 20mg / L , Thrombin 10ku / L.

[0048] Reagent R2

[0049] Tris buffer 100mmol / L, BSA 5g / L, PVP 30g / L, Tween-20 1ml / L, sodium azide 1g / L, HD-Phe-Pip-Arg-pNA·2HCl 2g / L.

Embodiment 3

[0051] The antithrombin III assay kit of this embodiment consists of the following components:

[0052] Reagent R1

[0053] Tris buffer 100mmol / L, BSA 1g / L, gelatin 30g / L, mannitol 30g / L, ammonium sulfate 1mol / L, Triton-100 1ml / L, sodium azide 1g / L, trehalose 50g / L , heparin 15mg / L, thrombin 10ku / L.

[0054] Reagent R2

[0055] Tris buffer 100mmol / L, BSA 5g / L, casein 5g / L, Tween-20 1ml / L,

[0056] Sodium azide 1g / L, HD-Phe-Pip-Arg-pNA·2HCl 2g / L.

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Abstract

The invention belongs to the technical field of biomedical examination, and relates to an antithrombin III determination kit and a detection method thereof. The kit comprises a reagent R1 and a reagent R2, wherein the reagent R1 comprises a buffer solution, a stabilizer, a salt, a surfactant, a preservative, trehalose, heparin and thrombin; and the reagent R2 comprises the buffer solution, the stabilizer, the surfactant, the preservative and a chromogenic substrate. A sample and deionized water are mixed according to a volume ratio of 1:(3-5) to obtain a diluted sample; and the diluted sampleis incubated for 30-60 s, the reagent R1 is added, uniform mixing and incubation are carried out for 60-100 s, the reagent R2 is added, uniform mixing is carried out, and then the obtained mixture isdetected at a wavelength of 405 nm.

Description

technical field [0001] The invention belongs to the technical field of biomedical testing, and relates to an antithrombin III assay kit and a detection method thereof. Background technique [0002] Antithrombin Ⅲ (AT-Ⅲ) is a single-chain glycoprotein, which belongs to α2 globulin. AT-Ⅲ has the function of decomposing lipoproteins and inhibits almost all active serine proteases in the coagulation system. AT-III, as the most important inhibitor of active coagulation factors in blood, controls blood coagulation and fibrin dissolution. The level of antithrombin III in the blood varies depending on various diseases and symptoms. Reduced in disseminated intravascular coagulation (DIC), liver disease, nephrotic syndrome, etc. Decreased levels of antithrombin III in the blood may prevent the effect of heparin therapy. Therefore, it is of great significance to know the activity of antithrombin III as an index for the monitoring, pathological analysis, prognosis judgment and hepari...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N21/31
CPCG01N21/31
Inventor 王卫华谢蒙张闻陈媛周海滨
Owner NINGBO RUI BIO TECH
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