Antithrombin activity assay reagent and preparation method and application thereof

Abstract

The invention discloses a reagent for detecting antithrombin activity, including reagent 1 and reagent 2, wherein: reagent 1 is a chromogenic substrate of FXa, and reagent 2 is a diluent containing FXa and heparin; wherein the chromogenic substrate of FXa is selected from From the following: CH 3 SO 2 ‑D‑Leu‑Gly‑Arg‑p‑nitroanilide·AcOH; CH 3 OCO‑D‑CHG‑Gly‑Arg‑p‑nitroanilide·AcOH; CH 3 OCO‑D‑CHA‑Gly‑Arg‑p‑nitroanilide AcOH; Benzoyl‑Ile‑Glu‑Gly‑Arg‑p‑nitroanilide HCl; Suc‑Ile‑Glu(γ‑Piperidyl)‑Gly‑Arg‑p‑nitroanilide HCl; N-α-Z-D-Arg-Gly-Arg-p-nitroanilide 2HCl; Boc-D-Arg-Gly-Arg-p-nitroanilide 2HCl; Acetyl-D-Arg-Gly-Arg-p ‑nitroanilide·2HCl; 4‑Nz‑D‑Arg‑Gly‑Arg‑p‑nitroanilide·2HCl; 4‑Mbs‑D‑Arg‑Gly‑Arg‑p‑nitroanilide·2HCl. The invention has good detection stability and repeatability, high sensitivity and accuracy, can accurately reflect the activity of antithrombin, and can find patients with abnormal antithrombin activity in the body, thereby providing a basis for treatment.

Description

technical field [0001] The invention belongs to the technical field of biological detection, and relates to an antithrombin activity detection reagent and a preparation method and application thereof. Background technique [0002] 1. Human body's hemostasis (coagulation) mechanism [0003] The hemostatic (coagulation) mechanism is a very important self-protection function of the human body. The hemostatic (coagulation) mechanism includes two aspects of coagulation and anticoagulation. The dynamic balance between the two is the normal body to maintain the state of blood flow in the body and prevent blood loss. The key is that it can close the wound of blood vessel through the aggregation of platelets when the blood vessel is injured and damaged, so as to prevent a large amount of blood loss. The hemostasis (coagulation) process is a process in which a series of coagulation factors are activated by enzymatic hydrolysis, and the hemostasis (coagulation) process finally generat...

AI Technical Summary

Problems solved by technology

Therefore, the scope of application of this method is limited

[0012] 4. Limitations of Existing Technologies

However, because it uses heparin derivatives and imported thrombin, and the heparin derivatives need to be obtained from the cracking of heparin, this increases the cost of reagent production, and also makes the steps of reagent production more cumbersome.

Due to the high cost of this technology, the amount of reagents is reduced, and the reduction in volume may cause some values ​​​​not to be displayed in the final colorimetric process.

And this prior art needs to change a lot of parameters when carrying out ATIII activity detection on the fully automatic coagulation instrument, and operation is inconvenient, and the result is also prone to error

[0014] In addition, Chinese invention patent application CN104714036A also discloses an antithrombin III activity assay kit, which also uses a thrombin-based chromogenic substrate method for determination, but the thrombin-based chromogenic substrate There are serious flaws in the

This method analyzes the activity of antithrombin in the plasma to be tested by adding exogenous thrombin, because another thrombin inhibitor other than antithrombin naturally exists in the plasma to be tested—heparin cofactor II (HCII ), so this method is susceptible to the interference of HCII in the plasma to be tested, which leads to inaccurate determination of antithrombin

Because heparin cofactor II is naturally present in the plasma of the general population, antithrombin activity measured by this method is inaccurate

What's more worth mentioning is that this method cannot be used for people who take Hirudin, Argatroban or Dabigatran, because these drugs are direct inhibitors of thrombin and will High antithrombin activity test results

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