Protein C activity determination kit based on chromophoric substrate method

Abstract

The invention discloses a protein C activity determination kit based on a chromophoric substrate method, protein C is a vitamin K dependent plasma serine protease zymogen, and comprises a reagent R1, a reagent R2 and a diluting reagent; the R1 reagent comprises a protein C activator, a first buffer solution and a first auxiliary material; the R2 reagent comprises a chromogenic substrate Pca-5297, a second buffer solution and a second auxiliary material; the diluting reagent comprises a third buffer solution and a third auxiliary material, and the pH of the first buffer solution, the pH of the second buffer solution and the pH of the third buffer solution are 7.2-7.6; the protein C activator and the chromophoric substrate Pca-5297 are matched for use, during detection, the cutting efficiency is high, the reaction signal is strong, the reaction speed is high, the sensitivity is high, the linear range is wide, the reaction time is short, the sample distinction degree is increased, and establishment of the linear range and clinical sample testing are facilitated; the reagent is liquid, convenient to use, low in cost and good in stability.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a protein C activity assay kit based on a chromogenic substrate method. Background technique [0002] Protein C (referred to as PC) is a vitamin K-dependent plasma serine protease zymogen, mainly synthesized in the liver, which is converted into activated protein C (referred to as APC) under the action of thrombin or thrombin-thrombomodulin complex. ). APC forms a complex with its cofactor protein S (referred to as PS), which has the activity of inactivating Va and VIIIa and increasing fibrinolysis, so it has anticoagulant effect. When PC is deficient, the inactivation of factors Va and VIIIa is reduced and the fibrinolytic capacity of blood circulation is reduced, thus promoting excessive fibrin formation and leading to thrombosis. Therefore, the determination of PC plays an important role in disease prevention, monitoring and treatment. . [0003] At present, the methods for qua...

AI Technical Summary

Problems solved by technology

Among them, although the antigen-antibody reaction in immunological methods can accurately detect the content of PC antigen, it requires highly purified PC for the screening of specific antibodies, and this condition is very harsh; secondly, the entire detection process takes a long time, Unable to meet the needs of emergency patients and clinical timely diagnosis

The APTT coagulation method uses snake venom to activate PC, and then adds it to the plasma to be tested. The content of PC corresponds to the prolongation of the plasma coagulation time. This method can specifically detect the content of protein C, but there are many factors that affect the results. , the experience of inspectors often directly affects the accuracy and repeatability of test results

In the early stage of the chromogenic substrate method, thrombin-thrombin regulatory protein was used as the activator, but because of the presence of PC inhibitors and interfering substances in the plasma, it cannot directly detect the PC activity in the plasma, but needs to separate the PC from the plasma. This will not only waste a lot of manpower and material resources, but also slow the detection speed

[0004] Most of the PC activity assay kits (chromogenic substrate method) on the domestic market are mainly imported kits, which are expensive, have a long procurement cycle, and are mostly in the form of freeze-dried powder. The production process is cumbersome and the cost is high. The difference between the bottles will also be enlarged due to freeze-drying. The freeze-dried powder reagents need to be reconstituted before use, and can be used after standing for a period of time, which is not conducive to the customer's operation; this type of PC activity assay kit, there is no A reconstitution solvent is included, and the quality of the reconstitution solvent selected by the customer cannot be controlled, which has a certain impact on the test results; the sensitivity of the reagent is poor, the linear range is narrow, and the specificity is poor, resulting in poor repeatability of the test results and anti-interference Poor ability, the results are easily affected by various factors, and cannot reflect the real protein C activity level in the human body

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