阅读说明：本技术 一种血小板抑制率检测试剂盒及其制备方法和检测方法 (A kind of blood platelet inhibiting rate detection kit and preparation method thereof and detection method ) 是由 陈文聪 肖良品 余枝广 卢景江 胡维 吴林涛 于 2019-08-28 设计创作，主要内容包括：本发明提供了一种血小板抑制率检测试剂盒,包括枸橼酸全血激活剂、第一冻干粉和第二冻干粉,所述第一冻干粉包括纤维蛋白原激活剂冻干粉和血小板抑制剂冻干粉,所述第二冻干粉包括血小板激活剂-二价阳离子复合物冻干粉和所述纤维蛋白原激活剂冻干粉；其中,第一冻干粉中的血小板抑制剂可以有效避免由于外界因素导致血小板激活的现象；第二冻干粉中直接混合有纤维蛋白原激活剂和血小板激活剂,提高检测效率,同时,血小板激活剂-二价阳离子复合物可以有效避免血小板激活剂和纤维蛋白原激活剂之间形成复合物,保证了血小板激活剂活性和稳定性,提高检测准确性。本发明还提供了上述血小板抑制率检测试剂盒的制备方法和检测方法。(The present invention provides a kind of blood platelet inhibiting rate detection kits, including citric acid whole blood activator, the first freeze-dried powder and the second freeze-dried powder, first freeze-dried powder includes fibrinogen activator freeze-dried powder and platelet suppressant drug freeze-dried powder, and second freeze-dried powder includes platelet activating agent-bivalent cation compound freeze-dried powder and the fibrinogen activator freeze-dried powder；Wherein, the platelet suppressant drug in the first freeze-dried powder is it is possible to prevente effectively from the phenomenon that leading to platelet activation due to extraneous factor；Fibrinogen activator and platelet activating agent are directly mixed in second freeze-dried powder, improve detection efficiency, simultaneously, platelet activating agent-bivalent cation compound is it is possible to prevente effectively from form compound between platelet activating agent and fibrinogen activator, it ensure that platelet activating agent activity and stability, improve detection accuracy.The present invention also provides the preparation methods and detection method of above-mentioned blood platelet inhibiting rate detection kit.)

1. a kind of blood platelet inhibiting rate detection kit, which is characterized in that including citric acid whole blood activator, the first freeze-dried powder and Second freeze-dried powder, first freeze-dried powder include fibrinogen activator freeze-dried powder and platelet suppressant drug freeze-dried powder, described Second freeze-dried powder includes platelet activating agent-bivalent cation compound freeze-dried powder and fibrinogen activator freeze-drying Powder.

2. blood platelet inhibiting rate detection kit as described in claim 1, which is characterized in that the platelet activating agent-two Bivalent cation includes Zn in valence cationic compound freeze-dried powder²⁺、Mg²⁺、Ca²⁺And Fe²⁺At least one of.

3. blood platelet inhibiting rate detection kit as described in claim 1, which is characterized in that the platelet activating agent-two In valence cationic compound freeze-dried powder platelet activating agent-bivalent cation compound by platelet activating agent and divalent sun from Son chemical bonding is formed.

4. blood platelet inhibiting rate detection kit as claimed in claim 3, which is characterized in that the platelet activating agent includes At least one of arachidonic acid and its sodium salt, sylvite.

5. blood platelet inhibiting rate detection kit as described in claim 1, which is characterized in that described in first freeze-dried powder The mass content of platelet suppressant drug freeze-dried powder is 0.001%-10%.

6. blood platelet inhibiting rate detection kit as described in claim 1, which is characterized in that first freeze-dried powder uniformly divides It is scattered at least one first reaction cup, second freeze-dried powder is evenly spread at least one second reaction cup.

7. blood platelet inhibiting rate detection kit as described in claim 1, which is characterized in that the fibrinogen activator Freeze-dried powder includes Batroxobin and activated clotting factor, and the activated clotting factor includes activated clotting factor V, activated clotting factor At least one of VIII, activated clotting factor X, activated clotting factor XIII and activated clotting factor XIIIa.

8. blood platelet inhibiting rate detection kit as described in claim 1, which is characterized in that the platelet suppressant drug freeze-drying Powder include aspirin, tirofiban, Plavix, kikri obtain, prasugrel, ticagrelor, cangrelor, eptifibatide At least one of with platelet antibody.

9. a kind of preparation method of blood platelet inhibiting rate detection kit characterized by comprising

The first freeze-dried powder is provided, first freeze-dried powder includes fibrinogen activator freeze-dried powder and platelet suppressant drug freeze-drying Powder；

The second freeze-dried powder is provided, second freeze-dried powder includes platelet activating agent-bivalent cation compound freeze-dried powder and institute State fibrinogen activator freeze-dried powder；

The activator of citric acid whole blood is provided, blood platelet inhibiting rate detection kit can be obtained.

10. a kind of detection method using such as the described in any item blood platelet inhibiting rate detection kits of claim 1-8, special Sign is, comprising:

Citrate anticoagulation whole blood and anticoagulant heparin whole blood are acquired respectively；

Citrate anticoagulation whole blood injection is contained into the citric acid whole blood activator, measures clot strength M₁；

The anticoagulant heparin whole blood is injected into the reaction cup containing first freeze-dried powder, clot strength M is measured₂；

The anticoagulant heparin whole blood is injected into the reaction cup containing second freeze-dried powder, clot strength M is measured₃；

Blood platelet inhibiting rate is obtained according to the following formula:

Technical field

The present invention relates to medical domains, and in particular to a kind of blood platelet inhibiting rate detection kit and preparation method thereof and inspection Survey method.

Background technique

Blood platelet is in physiological haemostasis, maintenance blood vessel wall integrity and certain pathologic processes, such as thrombosis, artery congee It plays an important role during sample hardening, unstable angina, metastases and inflammatory reaction etc..A large number of studies show that blood Activation, aggregation, inhibition of platelet etc. are the initiating agents of internal thrombosis, are also the compositions of most critical in thrombosis One of.Thrombus elasticity drawing method is the method that clinically can commonly quantify to detect platelet function at present.Thrombelastogram The special graph that instrument is depicted can completely monitor blood sample and be formed from blood coagulation to blood clot and the full mistake of fibrinolysis Journey is realized and carries out blood coagulation overall picture to coagulation factor, fibrinogen, platelet aggregation and fibrinolysis etc. Testing and evaluation, be widely used in department of cardiac surgery, organ transplant, tumour and radiotherapy or the postoperative hemorrhage in later period and The case where thrombosis, monitors.

Currently, blood platelet inhibiting rate detection process needs to be added the blood coagulation of blood after citric acid whole blood activator by detection Intensity, clot strength, addition fibrinogen activator and the platelet activating agent that blood after fibrinogen activator is added The clot strength of blood afterwards, and then obtain blood platelet inhibiting rate.This process fibrinogen activator and platelet activating agent storage There are in cillin bottle, it is lyophilized powder, needs to be redissolved when in use, and be transferred in reaction cup, it is time-consuming and laborious and take Amount has great error；Operating procedure is many and diverse, in a large amount of detections, is easy to add reagent on mistake in adding procedure；Meanwhile Can also there be the shearing force in the case where blood platelet is activated by other extraneous factors, such as low temperature, detection process, to addition fiber The clot strength detection of blood produces bigger effect after albumen activator, so that detection intensity M2 is apparently higher than actual strength, And then influence the assessment of blood platelet inhibiting rate.

Summary of the invention

To solve the above problems, the present invention provides a kind of blood platelet inhibiting rate detection kit, in the first freeze-dried powder Platelet suppressant drug is it is possible to prevente effectively from the phenomenon that leading to platelet activation due to extraneous factor, so that detection intensity M2 and reality Intensity is consistent, improves the accuracy of detection；Fibrinogen activator and platelet activation are directly mixed in second freeze-dried powder Agent avoids redissolution and mixing in operation, saves the operating time, improves efficiency and accuracy, while fibrinogen activator Directly mixing with platelet activating agent will form compound, reduce the activity of platelet activating agent, platelet activating agent-divalent sun Ion complex ensure that blood is small it is possible to prevente effectively from form compound between platelet activating agent and fibrinogen activator Plate activator activity, improves detection accuracy.

In a first aspect, the present invention provides a kind of blood platelet inhibiting rate detection kit, including citric acid whole blood activator, First freeze-dried powder and the second freeze-dried powder, first freeze-dried powder include fibrinogen activator freeze-dried powder and platelet suppressant drug Freeze-dried powder, second freeze-dried powder include platelet activating agent-bivalent cation compound freeze-dried powder and the fibrinogen Activator freeze-dried powder.

In the present invention, platelet suppressant drug is it is possible to prevente effectively from the phenomenon that leading to platelet activation due to extraneous factor； Directly be mixed with fibrinogen activator and platelet activating agent in second freeze-dried powder, save operating procedure, improve efficiency and Accuracy；Simultaneously as fibrinogen activator and platelet activating agent, which directly mix, will form compound, to reduce blood Platelet activator activity, platelet activating agent-bivalent cation compound is it is possible to prevente effectively from platelet activating agent and fiber egg Compound is formed between white activator, platelet activating agent activity is ensure that, improves detection accuracy.

Optionally, bivalent cation includes Zn in the platelet activating agent-bivalent cation compound freeze-dried powder²⁺、Mg^{2 +}、Ca²⁺And Fe²⁺At least one of.

Optionally, platelet activating agent includes peanut in the platelet activating agent-bivalent cation compound freeze-dried powder At least one of four diluted acids and its sodium salt, sylvite.

Optionally, in the platelet activating agent-bivalent cation compound freeze-dried powder platelet activating agent-divalent sun from Sub- compound is chemically bonded to be formed by platelet activating agent and bivalent cation.

Optionally, the platelet activating agent includes at least one of arachidonic acid and its sodium salt, sylvite, blood platelet Activator-bivalent cation compound effectively avoids being formed between arachidonic acid and its natrium potassium salt and fibrinogen activator Compound ensure that the activity of arachidonic acid and its natrium potassium salt, improve detection accuracy.That is, the second freeze-dried powder packet Include the freeze-dried powder and the fibre of the compound that at least one of arachidonic acid and its sodium salt, sylvite are formed with bivalent cation Fibrillarin activator freeze-dried powder.

Optionally, the mass content of platelet suppressant drug freeze-dried powder described in first freeze-dried powder is 0.001%- 10%.Further, the mass content of platelet suppressant drug freeze-dried powder described in first freeze-dried powder is 0.01%-9%.More Further, the mass content of platelet suppressant drug freeze-dried powder described in first freeze-dried powder is 0.1%-8%.

Optionally, first freeze-dried powder is evenly spread at least one first reaction cup, and second freeze-dried powder is equal It is even to be distributed at least one second reaction cup.In the present invention, the first freeze-dried powder and the second freeze-dried powder are dispensed to reaction cup In, the phenomenon that the solvent and blood of redissolution can directly be added in reaction cup in operation, reagent loss is not present, and institute There is operation to carry out directly in reaction cup, streamline operation.

Optionally, the content of the first freeze-dried powder described in each first reaction cup is 1 μ g-300 μ g.Further, The content of first freeze-dried powder described in each first reaction cup is 10 μ g-230 μ g.Further, each described first The content of first freeze-dried powder described in reaction cup is 20 μ g-150 μ g.

Optionally, the content of the second freeze-dried powder described in each second reaction cup is 1 μ g-300 μ g.Further, The content of second freeze-dried powder described in each second reaction cup is 10 μ g-230 μ g.Further, each described second The content of second freeze-dried powder described in reaction cup is 20 μ g-150 μ g.

Optionally, the fibrinogen activator freeze-dried powder includes Batroxobin and activated clotting factor, and the activation is solidifying Blood factor includes activated clotting factor V, activated clotting factor VIII, activated clotting factor X, activated clotting factor XIII and activation At least one of factor XIII a.

Optionally, the platelet suppressant drug freeze-dried powder includes that aspirin, tirofiban, Plavix, kikri obtain, are general At least one of glug thunder, ticagrelor, cangrelor, eptifibatide and platelet antibody.

Optionally, the citric acid whole blood activator includes electronegative compound and calcium chloride.Further, the band The compound of negative electricity includes at least one of kaolin, white bole, ellagic acid and collagen.

It optionally, further include third freeze-dried powder, the third freeze-dried powder includes the fibrinogen activator freeze-dried powder With at least one of adenosine diphosphate (ADP) and its sodium salt, sylvite freeze-dried powder.It in the present invention, can when including third freeze-dried powder With the two kinds of approach inhibited to blood platelet while assessing.Further, the third freeze-dried powder is evenly dispersed at least one In a third reaction cup.More further, the content of third freeze-dried powder described in each third reaction cup is 1 μ g-300 μ g。

The blood platelet inhibiting rate detection kit that first aspect present invention provides, including citric acid whole blood activator, first Freeze-dried powder and the second freeze-dried powder, the first freeze-dried powder include fibrinogen activator freeze-dried powder and platelet suppressant drug freeze-dried powder, Second freeze-dried powder includes platelet activating agent-bivalent cation compound freeze-dried powder and fibrinogen activator freeze-dried powder；Blood Platelet inhibitor is it is possible to prevente effectively from the phenomenon that leading to platelet activation due to extraneous factor；It is directly mixed in second freeze-dried powder Fibrinogen activator and platelet activating agent improve detection efficiency, meanwhile, platelet activating agent-bivalent cation is compound Object ensure that platelet activating agent it is possible to prevente effectively from form compound between platelet activating agent and fibrinogen activator Activity improves detection accuracy.

Second aspect, the present invention provides a kind of preparation methods of blood platelet inhibiting rate detection kit, comprising:

The first freeze-dried powder is provided, first freeze-dried powder includes fibrinogen activator freeze-dried powder and platelet suppressant drug Freeze-dried powder；

The second freeze-dried powder is provided, second freeze-dried powder includes platelet activating agent-bivalent cation compound freeze-dried powder With the fibrinogen activator freeze-dried powder；

Citric acid whole blood activator is provided, blood platelet inhibiting rate detection kit can be obtained.

Optionally, fibrinogen activator and freeze-dried formation first freeze-dried powder of platelet suppressant drug mixing, or The fibrinogen activator is lyophilized to form the fibrinogen activator freeze-dried powder, and shape is lyophilized in the platelet suppressant drug At the platelet suppressant drug freeze-dried powder, the fibrinogen activator freeze-dried powder and the platelet suppressant drug freeze-dried powder are mixed Conjunction forms first freeze-dried powder.

Optionally, platelet activating agent and solution containing bivalent cation mixing after formed platelet activating agent-divalent sun from Sub- compound, then mixed with the fibrinogen activator, it is freeze-dried to form second freeze-dried powder or platelet activating agent Platelet activating agent is formed with after the mixing of solution containing bivalent cation, freeze-dried formation platelet activating agent-bivalent cation is multiple Object freeze-dried powder is closed, the fibrinogen activator is lyophilized to form the fibrinogen activator freeze-dried powder, the blood platelet Activator-bivalent cation compound freeze-dried powder and the fibrinogen activator freeze-dried powder are mixed to form second freeze-drying Powder.

Optionally, the concentration of the solution containing bivalent cation is 0.0001mM-100mM.Further, described to contain divalent The concentration of cationic solution is 0.001mM-98mM.Further, the concentration of the solution containing bivalent cation is 0.01mM- 90mM。

It optionally, include Zn in the solution containing bivalent cation²⁺、Mg²⁺、Ca²⁺And Fe²⁺At least one of.

The blood platelet inhibiting rate detection reagent box preparation method that second aspect of the present invention provides is simple, can be used for extensive life It produces and applies

The third aspect, the present invention provides a kind of blood platelet inhibiting rate detection kits used as described in relation to the first aspect Detection method, comprising:

Citrate anticoagulation whole blood and anticoagulant heparin whole blood are acquired respectively；

Citrate anticoagulation whole blood injection is contained into the citric acid whole blood activator, measures clot strength M₁；

The anticoagulant heparin whole blood is injected into the reaction cup containing first freeze-dried powder, clot strength M is measured₂；

The anticoagulant heparin whole blood is injected into the reaction cup containing second freeze-dried powder, clot strength M is measured₃；

Blood platelet inhibiting rate is obtained according to the following formula:

The detection method for the blood platelet inhibiting rate detection kit that third aspect present invention provides, easy to operate, simplified inspection Flow gauge, detection accuracy are excellent.

To sum up, beneficial effect of the present invention includes the following aspects:

1, in blood platelet inhibiting rate detection kit provided by the invention, platelet suppressant drug in the first freeze-dried powder can be with Effectively avoid the phenomenon that platelet activation is led to due to extraneous factor；Fibrinogen activation is directly mixed in second freeze-dried powder Agent and platelet activating agent are saved operation, are improved efficiency and accuracy, while fibrinogen activator and platelet activating agent Directly mixing will form compound, influence the activity of platelet activating agent, and platelet activating agent-bivalent cation compound can be with It effectively avoids forming compound between platelet activating agent and fibrinogen activator, ensure that platelet activating agent activity, Improve detection accuracy；

2, blood platelet inhibiting rate detection reagent box preparation method provided by the invention is simple, can be used for being mass produced and answering With；

3, the detection method of blood platelet inhibiting rate detection kit provided by the invention, easy to operate, simplified testing process, Detection accuracy is excellent.

Detailed description of the invention

In order to more clearly explain the embodiment of the invention or the technical proposal in the existing technology, to embodiment or will show below There is attached drawing needed in technical description to be briefly described.Specific embodiment described herein is only used to explain this Invention, is not intended to limit the present invention.

Fig. 1 is the result figure of the clot strength of effect example 1 of the present invention detection.

Fig. 2 is the result figure of the clot strength of effect example 2 of the present invention detection.

Specific embodiment

The following is a preferred embodiment of the present invention, it is noted that for those skilled in the art For, various improvements and modifications may be made without departing from the principle of the present invention, these improvements and modifications are also considered as Protection scope of the present invention.

The present invention provides a kind of blood platelet inhibiting rate detection kits, including citric acid whole blood activator, the first freeze-drying Powder and the second freeze-dried powder, the first freeze-dried powder include the freeze-dried powder that fibrinogen activator and platelet suppressant drug are mixed with, Second freeze-dried powder includes the freeze-drying that platelet activating agent-bivalent cation compound and fibrinogen activator is mixed with Powder.

It is such as low the case where activation there are blood platelet by other extraneous factors in existing blood platelet inhibiting rate detection process Shearing force etc. in temperature, detection process, detection intensity is apparently higher than actual strength, Jin Erying after fibrinogen activator is added Ring the assessment of blood platelet inhibiting rate.The addition of platelet suppressant drug is it is possible to prevente effectively from since extraneous factor leads to platelet activation The phenomenon that；Also, fibrinogen activator and platelet suppressant drug are mixed and made into freeze-dried powder, improve efficiency.But it is existing In blood platelet inhibiting rate detection process, due to fibrinogen activator also need it is strong with platelet activating agent hybrid detection blood coagulation Degree, directly adds platelet suppressant drug, and will affect fibrinogen activator and blood platelet in fibrinogen activator Therefore the clot strength of activator hybrid detection individually adds platelet suppressant drug when detecting, will increase operating process.This In invention, fibrinogen activator and platelet suppressant drug are mixed to prepare the first freeze-dried powder, both will not influence fibrinogen The clot strength detected when activator is mixed with platelet activating agent in turn avoids the phenomenon that blood platelet is because of other factors activation, Detection accuracy improves.

It will form compound since fibrinogen activator and platelet activating agent directly mix, influence platelet activation Agent activity；Platelet activating agent-bivalent cation compound is it is possible to prevente effectively from platelet activating agent and fibrinogen activation Compound is formed between agent, be ensure that platelet activating agent activity, is improved detection accuracy.

In the embodiment of the present invention, bivalent cation includes Zn in platelet activating agent-bivalent cation compound freeze-dried powder^{2 +}、Mg²⁺、Ca²⁺And Fe²⁺At least one of.

In the embodiment of the present invention, platelet activating agent-divalent in platelet activating agent-bivalent cation compound freeze-dried powder Cationic compound is chemically bonded to be formed by platelet activating agent and bivalent cation.Optionally, platelet activating agent includes At least one of arachidonic acid and its sodium salt, sylvite.Platelet activating agent includes arachidonic acid, arachidonic acid sodium At least one of salt, arachidonic acid sylvite.

In the embodiment of the present invention, the mass content of platelet suppressant drug is 0.001%-10% in the first freeze-dried powder.Into one Step, the mass content of platelet suppressant drug is 0.01%-9% in the first freeze-dried powder.Further, blood in the first freeze-dried powder The mass content of platelet inhibitor is 0.1%-8%.Specifically, the mass content of platelet suppressant drug can be in the first freeze-dried powder But it is not limited to 0.0058%, 0.073%, 0.28%, 3%, 4.9% or 6.1%.

In the embodiment of the present invention, the first freeze-dried powder is evenly spread at least one first reaction cup, and the second freeze-dried powder is equal It is even to be distributed at least one second reaction cup.In the present invention, the first freeze-dried powder and the second freeze-dried powder are dispensed to reaction cup In, the phenomenon that the solvent and blood of redissolution can directly be added in reaction cup in operation, reagent loss is not present, and institute There is operation to carry out directly in reaction cup, streamline operation.

In the embodiment of the present invention, the first reaction cup is different from the color of the second reaction cup, to distinguish.

In the embodiment of the present invention, the content of the first freeze-dried powder is 1 μ g-300 μ g in every one first reaction cup.Further, The content of the first freeze-dried powder is 10 μ g-230 μ g in every one first reaction cup.Further, first in every one first reaction cup The content of freeze-dried powder is 20 μ g-150 μ g.Specifically, the content of the first freeze-dried powder can be, but not limited in every one first reaction cup For 3.5 μ g, 18.2 μ g, 90.7 μ g, 178.4 μ g or 261 μ g.

In the embodiment of the present invention, the content of the second freeze-dried powder is 1 μ g-300 μ g in every one second reaction cup.Further, The content of the second freeze-dried powder is 10 μ g-230 μ g in every one second reaction cup.Further, second in every one second reaction cup The content of freeze-dried powder is 20 μ g-150 μ g.Specifically, the content of the second freeze-dried powder can be, but not limited in every one second reaction cup For 5 μ g, 22 μ g, 130 μ g, 195 μ g or 270 μ g.

In the embodiment of the present invention, fibrinogen activator freeze-dried powder includes Batroxobin and activated clotting factor, the work Change coagulation factor include activated clotting factor V, activated clotting factor VIII, activated clotting factor X, activated clotting factor XIII and At least one of activated clotting factor XIIIa.Specifically, fibrinogen activator freeze-dried powder can be, but not limited to include bar Inulinase and activated clotting factor XIIIa.

In the embodiment of the present invention, platelet suppressant drug freeze-dried powder includes aspirin, tirofiban, Plavix, kikri , at least one of prasugrel, ticagrelor, cangrelor, eptifibatide and platelet antibody.Wherein, Ah Si Woods (Aspirin) is acetylsalicylic acid；Tirofiban (Tirofiban) chemical name: N- (butyl sulfonyl)-O- [4- (4- piperazine Piperidinyl) butyl]-l-tyrosine；Plavix (clopidogrel bisulfate tablet) chemical name: methyl (+)-(s)-α-Chloro-O-Phenyl- 6,7- dichloro-thiophene [3,2-C] piperidines -5 (4H)-ethyl acetate disulfate；Kikri obtains (Assay of ticlopidine hydrochloride tablet) chemical name Claim: 5- [(2- chlorphenyl) methyl] -4,5,6,7- thiophane simultaneously [3,2-C] pyridine hydrochloride；Prasugrel chemical name: 2- [2- (acetoxyl group) -6,7- dihydro-thiophene simultaneously [3,2-c] pyridine -5 (4H)-yl] -1- cyclopropyl -2- (2- fluorophenyl) ethyl ketone； Platelet antibody includes monoclonal antibody and at least one of more anti-, specifically can be, but not limited to Abciximab；Ticagrelor chemistry Title: (1S, 2S, 3R, 5S) -3- [7- [[(1R, 2S) -2- (3,4- difluorophenyl) cyclopropyl] amino] -5- rosickyite base triazol [4,5-d] pyrimidin-3-yl] -5- (2- hydroxy ethoxy) -1,2- cyclopentadienyl alcohol；Eptifibatide molecular formula C₃₅H₄₉N₁₁O₉S₂。

In the embodiment of the present invention, citric acid whole blood activator includes electronegative compound and calcium chloride.Further, band The compound of negative electricity includes at least one of kaolin, white bole, ellagic acid and collagen.

Further include third freeze-dried powder in the embodiment of the present invention, third freeze-dried powder include the fibrinogen activator and At least one of adenosine diphosphate (ADP) and its sodium salt, sylvite freeze-dried powder.It in the present invention, can be with when including third freeze-dried powder Two kinds of approach that blood platelet inhibits are assessed simultaneously.Further, third freeze-dried powder is evenly dispersed at least one third In reaction cup.More further, the content of third freeze-dried powder is 1 μ g-300 μ g in each third reaction cup.

Blood platelet inhibiting rate detection kit provided by the invention includes citric acid whole blood activator, the first freeze-dried powder and Two freeze-dried powders, the first freeze-dried powder include fibrinogen activator freeze-dried powder and platelet suppressant drug freeze-dried powder, the second freeze-dried powder Including platelet activating agent-bivalent cation compound freeze-dried powder and fibrinogen activator freeze-dried powder；Platelet suppressant drug It is possible to prevente effectively from the phenomenon that leading to platelet activation due to extraneous factor；Fibrinogen is directly mixed in second freeze-dried powder Activator and platelet activating agent improve detection efficiency, meanwhile, platelet activating agent-bivalent cation compound can be effective It avoids forming compound between platelet activating agent and fibrinogen activator, ensure that platelet activating agent activity, improve Detection accuracy.

The present invention also provides a kind of preparation methods of blood platelet inhibiting rate detection kit, comprising:

The first freeze-dried powder is provided, the first freeze-dried powder includes fibrinogen activator freeze-dried powder and platelet suppressant drug freeze-drying Powder；

The second freeze-dried powder is provided, the second freeze-dried powder includes platelet activating agent-bivalent cation compound freeze-dried powder and fibre Fibrillarin activator freeze-dried powder；

Citric acid whole blood activator is provided, blood platelet inhibiting rate detection kit can be obtained.

In the embodiment of the present invention, fibrinogen activator and freeze-dried the first freeze-drying of formation of platelet suppressant drug mixing Powder or fibrinogen activator are lyophilized to form fibrinogen activator freeze-dried powder, and platelet suppressant drug is lyophilized that form blood small Plate inhibitor freeze-dried powder, fibrinogen activator freeze-dried powder and platelet suppressant drug freeze-dried powder are mixed to form the first freeze-dried powder.

In the embodiment of the present invention, platelet activating agent-is formed after platelet activating agent and the mixing of solution containing bivalent cation Bivalent cation compound, then mixed with fibrinogen activator, the second freeze-dried powder of freeze-dried formation or platelet activating agent Platelet activating agent is formed with after the mixing of solution containing bivalent cation, freeze-dried formation platelet activating agent-bivalent cation is multiple Object freeze-dried powder is closed, fibrinogen activator is lyophilized to form fibrinogen activator freeze-dried powder, platelet activating agent-divalent sun Ion complex freeze-dried powder and fibrinogen activator freeze-dried powder are mixed to form the second freeze-dried powder.

In the embodiment of the present invention, the concentration of the solution containing bivalent cation is 0.0001mM-100mM.Further, contain divalent The concentration of cationic solution is 0.001mM-98mM.Further, the concentration of the solution containing bivalent cation is 0.01mM- 90mM.Specifically, the concentration of the solution containing bivalent cation can be, but not limited to for 0.005mM, 0.09mM, 0.33mM, 6.4mM, 25.1mM or 50mM.

In the embodiment of the present invention, solution containing bivalent cation includes Zn²⁺、Mg²⁺、Ca²⁺And Fe²⁺At least one of.

The present invention also provides a kind of detection methods using such as above-mentioned blood platelet inhibiting rate detection kit, comprising:

Citrate anticoagulation whole blood and anticoagulant heparin whole blood are acquired respectively；

The injection of citrate anticoagulation whole blood is contained into citric acid whole blood activator, measures clot strength M₁；

Anticoagulant heparin whole blood is injected into the reaction cup containing the first freeze-dried powder, clot strength M is measured₂；

Anticoagulant heparin whole blood is injected into the reaction cup containing the second freeze-dried powder, clot strength M is measured₃；

Blood platelet inhibiting rate is obtained according to the following formula:

The detection method of blood platelet inhibiting rate detection kit provided by the invention, easy to operate, simplified testing process, inspection It is excellent to survey accuracy.

In a specific embodiment of the invention, provide including kaolin and calcium chloride citric acid whole blood activator；There is provided the One freeze-dried powder, wherein fibrinogen activator is Batroxobin and activated clotting factor XIIIa, and platelet suppressant drug is that blood is small Plate antibody, platelet antibody mass content is 0.005% in the first freeze-dried powder, and weighs 5 the first freeze-dried powders of μ g and be placed in reaction cup In；It is mixed after arachidonic acid and calcium ions solution (0.001mM) mixing, then with Batroxobin, activated clotting factor XIIIa It is freeze-dried that the second freeze-dried powder is made, and weigh 5 the second freeze-dried powders of μ g and be placed in reaction cup.Acquire citrate anticoagulation whole blood and heparin Anticoagulated whole blood；The injection of citrate anticoagulation whole blood is contained into citric acid whole blood activator, measures clot strength M₁；Anticoagulant heparin is complete Blood is injected into the reaction cup containing the first freeze-dried powder, measures clot strength M₂；Anticoagulant heparin whole blood is injected into containing second In the reaction cup of freeze-dried powder, clot strength M is measured₃；Blood platelet inhibiting rate is obtained according to the following formula:

Effect example 1

The same Heparinised whole blood of acquisition is divided into two parts, portion is placed in room temperature and guarantees that it is not activated, another by its It is placed in 4 DEG C of refrigerators and is taken out after 60min, make sample by low-temp activation.Using be added to 0.005% platelet antibody inhibitor and The first freeze-dried powder for being not added with platelet suppressant drug tests above-mentioned two parts of samples respectively, records clot strength M₂, as a result such as Fig. 1 institute Show.After sample is by low-temp activation, fibrinogen activator is only added and measures clot strength M₂It is apparently higher than before not being activated Actual strength, seriously affected the assessment of blood platelet inhibiting rate.And blood platelet is added in fibrinogen activator and inhibits After agent, clot strength M is measured₂It is not much different with being measured before not being activated.The result shows that it is small that blood is added in the first freeze-dried powder Plate inhibitor can be very good to avoid to cause the erroneous judgement of inhibiting rate result because the factors such as low temperature activate sample.

Effect example 2

Experimental group: platelet activating agent arachidonic acid and 5mM calcium chloride are carried out to be mixed and made into freeze-dried powder, and and fiber Albumen activator freeze-dried powder is mixed to prepare the second freeze-dried powder.It takes 5 the second freeze-dried powders of μ g to be placed in reaction cup, mixes 0h respectively Test tube of hepari people whole blood sample is tested with 2h, records clot strength M₃。

Control group: by platelet activating agent arachidonic acid and fibrinogen activator mixture freeze-dried powder, 5 are equally taken μ g is placed in reaction cup, is tested test tube of hepari people whole blood sample in 0h and 2h respectively, is recorded clot strength M₃。

Testing result is as shown in Figure 2, it can be seen that clot strength M of the experimental group in test test tube of hepari people whole blood sample₂When, Detected value is not influenced by the time, and testing result variation is little.And the control group that calcium chloride is not added is then tested after mixing 2h Clot strength decline is obvious.Show to can be very good to avoid after calcium chloride is added in fibrinogen activator and platelet activating agent The case where platelet activating agent inactivates caused by formation compound between platelet activating agent and fibrinogen activator, protects Accuracy is demonstrate,proved.

The embodiments described above only express several embodiments of the present invention, and the description thereof is more specific and detailed, but simultaneously Limitations on the scope of the patent of the present invention therefore cannot be interpreted as.It should be pointed out that for those of ordinary skill in the art For, without departing from the inventive concept of the premise, various modifications and improvements can be made, these belong to guarantor of the invention Protect range.Therefore, the scope of protection of the patent of the invention shall be subject to the appended claims.