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Electrochemical method-based PT (Prothrombin Time) test card and preparation method thereof

A prothrombin time and electrochemical technology, which is applied in the field of preparation of prothrombin time test cards, can solve problems such as few reports, and achieve the effects of fast detection process, convenient large-scale application and good repeatability

Inactive Publication Date: 2017-05-17
北京乐普诊断科技股份有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] Based on the above experience, there are still few reports and patents on the detection of PT based on electrochemical methods.

Method used

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  • Electrochemical method-based PT (Prothrombin Time) test card and preparation method thereof
  • Electrochemical method-based PT (Prothrombin Time) test card and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] The preparation of embodiment 1 phospholipid solution

[0039] The detection function of the prothrombin time detection card prepared by the invention is mainly based on the function of tissue factor lipid in the components, and the tissue factor lipid is prepared from recombinant tissue factor and phospholipid. Among them, phospholipid is an important component that assists tissue factor to play a role, and is an important component that the prothrombin time detection card can complete the detection. Therefore, preparing a uniform and stable phospholipid solution is an important prerequisite for preparing the prothrombin time detection card.

[0040] Specifically, lyophilized phosphatidylcholine and phosphatidylserine were mixed (0.6 g) at a mass ratio of 2:1 and added to 1 L of the buffer solution, and magnetically stirred for 30 min at 30-40° C. to fully disperse and dissolve.

Embodiment 2

[0041] Example 2 Preparation of Tissue Factor Lipidate

[0042] Tissue factor lipid is the main biologically active component in the prothrombin time test card, which participates in the initiation of exogenous coagulation process. Specifically, rTF was dissolved in the phospholipid mixed solution prepared in Example 1, mixed at room temperature and ultrasonicated for 15 minutes (setting working time 3 s, intermittent time 2 s), to obtain a tissue factor lipid solution. The tissue factor esterified product was dialyzed in Tris buffer solution (10 mM) at pH 7.0 for 48 hours at room temperature.

Embodiment 3

[0043] Embodiment 3 point membrane technology

[0044] The prothrombin time detection card (electrochemical method) prepared by the present invention has many differences from the traditional prothrombin time kit, such as the state of the two active ingredients when they react with the blood sample to be tested. For the former, the active ingredient is evenly dispersed in the reaction area of ​​the conductive substrate in the form of a solid film; while for the latter, the reaction reagent exists in the form of liquid when participating in the reaction. In order to prepare a uniformly dispersed solid film on the conductive substrate, the specific implementation method of preparation is:

[0045] A 50 mM Tris buffer solution was prepared with a content of 50 mM sodium chloride, 10% wt% sodium alginate, 3.5 wt% glycine, 5.0 wt% PEG 8000, 0.8 wt% thimerosal, 4 mM calcium chloride, pH 7.0. The buffer solution and the dialyzed tissue factor lipid were mixed evenly at a volume rati...

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Abstract

The invention discloses an electrochemical method-based PT (Prothrombin Time) test card and a preparation method thereof. The electrochemical method-based PT test card is formed by an electric conduction substrate, a chemical reagent and the like, wherein the chemical reagent is specifically prepared from the main components of tissue factors, phospholipid and the like. The electrochemical method-based PT test card prepared by the invention is capable of realizing PT detection on fingertip blood or a blood sample which is not subjected to blood coagulation treatment, and has the advantages of high accuracy, good repeatability and the like; the preparation method is simple, has low technical requirements on equipment and operators, is strong in operability and has a wide application prospect in the fields of clinical examination and the like.

Description

technical field [0001] The invention relates to prothrombin time (Prothrombin Time, PT) detection in the field of clinical blood diagnosis, in particular to a preparation method of a prothrombin time test card. Background technique [0002] Coagulation function testing is an important clinical testing item, which is of great significance for the diagnosis of various clinical diseases. The determination of blood coagulation function usually includes 4 inspection contents, namely prothrombin time (PT), activated partial thromboplastin time (APTT), thrombin time (TT) and fibrinogen (FIB). Among them, prothrombin time (PT) is one of the main coagulation performance test indexes in clinical practice. [0003] PT can be used to monitor the dosage and efficacy of oral anticoagulants, related blood coagulation factors and inhibitors, and confirm the defects and inhibitors of related prothrombin and fibrinogen. Prolonged PT is common in vitamin K deficiency, DIC, and taking anticoa...

Claims

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Application Information

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IPC IPC(8): G01N27/26
CPCG01N27/26
Inventor 刘芳邱笑违余占江
Owner 北京乐普诊断科技股份有限公司
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