CN109884327A - The detection of functional fiber albumen activator and its application - Google Patents
The detection of functional fiber albumen activator and its application Download PDFInfo
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- CN109884327A CN109884327A CN201910235531.1A CN201910235531A CN109884327A CN 109884327 A CN109884327 A CN 109884327A CN 201910235531 A CN201910235531 A CN 201910235531A CN 109884327 A CN109884327 A CN 109884327A
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Abstract
The invention proposes functional fiber albumen activator, the functional fiber albumen activator includes: extrinsic coagulation activator, platelet suppressant drug, buffer salt and protective agent.Contain extrinsic coagulation activator and platelet suppressant drug in functional fiber albumen activator of the invention simultaneously; it can inhibit the aggregation of blood platelet while activating extrinsic coagulation mechanism; also, salt, protective agent is added simultaneously in activator, the function of ensureing blood coagulation can be played.Thrombelastogram instrument is carried out using the activator and activates hemostasis examination, fibrinogen content in blood sample can be accurately measured, be with a wide range of applications.
Description
Technical field
The present invention relates to technical field of in vitro diagnostic reagents, and in particular to the detection of functional fiber proteinogen and its answers
With more particularly to functional fiber albumen activator, preparation method and functional fiber proteinogen detection kit.
Background technique
Fibrinogen, that is, factor I, is a kind of dimer being made of α A, β B and v chain, molecular weight 3400000,
It is made of 2964 amino acid.Plasma content is 2.0~4.0g/L, and biological half life is 96~144h, is present in absorption blood
In slurry, fibrin can be changed into after fibrin ferment and hageman factor a effect.
Fibrinogen is a kind of glycoprotein synthesized by liver, the α being made of respectively 610,461 and 411 amino acid
α, β β, dimer composed by 6 peptide chains of γ γ, molecular weight 340000.
Fibrinogen can promote the aggregation of blood platelet, promote growth, proliferation and the contraction of smooth muscle and endothelial cell,
Increase viscosity of blood and peripheral resistance, cause endothelial cell damage, promotes collagen and nuclifort synthesis, chemotactic list
Core/macrophage promotes red blood cell adhesion and thrombosis to migrating under inner membrance.Therefore, it is in the morbidity of cardiovascular disease
With particularly significant effect.
Clinical data shows: 1. fibrinogen level increases in patient, and myocardial infarction or sudden death can occur for majority, and fine
Fibrillarin level is higher, and risk is also bigger；2. it is one for promoting incidence of atherosclerosis that plasma fibrinogen, which increases,
Key factor；3. fibrinogen increases, blood is in hypercoagulative state, and blood flow velocity slows down, and viscosity of blood increases, and is easy to produce
Green blood bolt；4. being often accompanied by fibrinogen level raising when hypertension, it is the major reason for inducing and aggravating hypertension；
5. fibrinogen and fat metabolism close relation；6. old, smoking, it is fat, stress, oral contraceptive and diabetes etc.
Internal fibrinogen level can be promoted to increase.
Thrombelastogram is the special graph for the blood sample condensation process described by thrombelastogram instrument, is using whole blood sample
It can examine, be not necessarily to specially treated blood sample, be a kind of hemorheology detection convenient, fast, accurate, that detection can be operated by bed
Method.Pass through the monitoring and analysis to thrombelastogram, it can be estimated that the Coagulability of patient assists clinician solidifying to patient
Blood situation makes accurate judgement.Thrombelastogram provide by blood coagulation start to fibrin formed, platelet aggregation, fiber egg
The white overall process information for linking to blood clot formation and fibrinolysis, to coagulation factor, fibrinogen, platelet aggregation
Function and fibrinolysis etc. carry out the testing and evaluation of blood coagulation overall picture, have true reduction blood coagulation overall picture, sensitivity
Degree is high, detection precisely, the features such as visual result and data are detailed, more conventional blood coagulation inspection can it is sensitiveer, more rapidly reflect blood coagulation
It is abnormal.Ischemic event ability of the thrombelastogram in the inhibition situation of anti-platelet function and prediction Anti-platelet therapy
No less than goldstandard turbidimetry, even more it is better than turbidimetry, while there is reagent unification, operational standardization, easy to spread excellent
Point.
However the method for existing detection fibrinogen is mostly detected with optical method, testing principle is according to solidifying
Gu turbidity changes to measure the content of its fibrinogen, vulnerable to the substance in blood sample per se with turbidity during
It influences, to influence the accuracy of result, while needing to pre-process sample in advance, increase in detection process by artificial
The possibility that factor influences.
Therefore, still have for the detection method of fibrinogen at present to be developed.
Summary of the invention
The present invention is directed to solve at least one the technical problems existing in the prior art at least to a certain extent.For this purpose,
The invention proposes a kind of functional fiber albumen activator, preparation method and functional fiber proteinogen detection reagents
Box.
It should be noted that the present invention is the following discovery based on inventor and completes:
When exciting intrinsic coagulation mechanism to carry out blood coagulation, the clot strength MA of thrombelastogram instrument detection is in fiber egg
It is realized under both Bai Yuanyu platelet aggregations collective effect.Therefore, it is impossible to be accurately judged to the work that the two is respectively played
With in turn, can not determining fibrinogen content.
In view of this, inventor compounds platelet suppressant drug by using extrinsic coagulation activator, can start simultaneously
Extrinsic coagulation access and inhibition platelet aggregation, clot strength MA only reflects fibrinogen blood coagulation situation at this time, into
The detection of row fibrinogen can detect fibrinogen content with accurate quantitative analysis.
For this purpose, in one aspect of the invention, the invention proposes a kind of functional fiber albumen activators.According to this
The embodiment of invention, the functional fiber albumen activator include: extrinsic coagulation activator, platelet suppressant drug, delay
Rush salt and protective agent.It is activated simultaneously containing extrinsic coagulation in functional fiber albumen activator according to an embodiment of the present invention
Agent, platelet suppressant drug can inhibit again the aggregation of blood platelet while activating extrinsic coagulation mechanism, also, activate
Salt, protective agent is added simultaneously in agent, the function of ensureing blood coagulation can be played.Thrombelastogram activation is carried out using the activator
Hemostasis examination can accurately measure fibrinogen content in blood sample, be with a wide range of applications.Also, in blood
It can be carried out in the matched reagent cup of bolt elastic force figure, not need to carry out other specially treateds, it is convenient and efficient.
According to an embodiment of the invention, the functional fiber albumen activator can also have following supplementary technology special
Sign:
According to an embodiment of the invention, the extrinsic coagulation activator is selected from tissue factor.
According to an embodiment of the invention, the concentration of the extrinsic coagulation activator is 0.1~1mg/mL.
According to an embodiment of the invention, the platelet suppressant drug is in Yi Feiba object, tirofiban and Abciximab
At least one of.
According to an embodiment of the invention, the concentration of the platelet suppressant drug is 1~20mg/ml.
According to an embodiment of the invention, the protective agent is in bovine serum albumin(BSA), casein and human serum albumins
At least one of.
According to an embodiment of the invention, protectant concentration is 1~20mg/mL.
According to an embodiment of the invention, including buffer and salinity in the buffer salt.
According to an embodiment of the invention, the content of the salinity is 0.1~1 mass %.
According to an embodiment of the invention, the concentration of the buffer is 10~100mmol/L, pH value is 6~8.
According to an embodiment of the invention, the salt is selected from least one of sodium chloride, potassium chloride and calcium chloride.
According to an embodiment of the invention, the buffer in Tris, HEPES, PBS, glycine and MOPS at least
One of.
According to an embodiment of the invention, the functional fiber albumen activator further comprises preservative.
According to an embodiment of the invention, the preservative is selected from Proclin300, potassium sorbate and P-hydroxybenzoic acid first
At least one of ester.
According to an embodiment of the invention, the concentration of the preservative is 0.01~0.1 mass %.
In another aspect of this invention, functional fiber albumen activator noted earlier is prepared the invention proposes a kind of
Method.According to an embodiment of the invention, the described method includes: preparing the buffer；Described in being added into the buffer
Salinity, protective agent, stir evenly, to obtain the first mixed liquor；The extrinsic coagulation is added into first mixed liquor
Activator stirs evenly, to obtain the second mixed liquor；The platelet suppressant drug is added into second mixed liquor, stirs
It mixes uniformly, to obtain third mixed liquor；And the preservative is added into the third mixed liquor, it stirs evenly, so as to
Obtain the fibrinogen activator.Functional fiber proteinogen obtained according to the method for the embodiment of the present invention as a result,
Activator can accurately measure fibrinogen content in blood sample, also, this method is simple and convenient for operation, be suitable for rule
Modelling production.
In still another aspect of the invention, the invention proposes a kind of kits.According to an embodiment of the invention, the reagent
Box includes mentioned-above functional fiber albumen activator.It as a result, can be with using kit according to an embodiment of the present invention
Accurately measure fibrinogen content in blood sample.
According to an embodiment of the invention, the kit further comprises calcium chloride solution.
According to an embodiment of the invention, the concentration of the calcium chloride solution is 0.1~1mol/L.
According to an embodiment of the invention, the kit further comprises specimen cup.
According to an embodiment of the invention, the kit is used for thrombelastogram instrument hemostasis examination.
Additional aspect and advantage of the invention will be set forth in part in the description, and will partially become from the following description
Obviously, or practice through the invention is recognized.
Detailed description of the invention
Above-mentioned and/or additional aspect of the invention and advantage will become from the description of the embodiment in conjunction with the following figures
Obviously and it is readily appreciated that, in which:
Fig. 1 is the fibrinogen thrombus elastic force figure detection provided according to one embodiment of the present of invention and contrast agent box
Testing result correlation analysis.
Specific embodiment
The embodiment of the present invention is described below in detail.The embodiments described below is exemplary, and is only used for explaining this hair
It is bright, and be not considered as limiting the invention.
It should be noted that term " first ", " second " are used for description purposes only, it is not understood to indicate or imply phase
To importance or implicitly indicate the quantity of indicated technical characteristic.Define " first " as a result, the feature of " second " can be with
Explicitly or implicitly include one or more of the features.Further, in the description of the present invention, unless otherwise saying
Bright, the meaning of " plurality " is two or more.
The invention proposes the detections of functional fiber albumen activator, preparation method and functional fiber proteinogen to try
Agent box will be described in greater detail respectively below.
Functional fiber albumen activator
In one aspect of the invention, the invention proposes a kind of functional fiber albumen activators.According to the present invention
Embodiment, the functional fiber albumen activator include: extrinsic coagulation activator, platelet suppressant drug, buffer salt and
Protective agent.Contain extrinsic coagulation activator and blood in functional fiber albumen activator according to an embodiment of the present invention simultaneously
Platelet inhibitor, while activating extrinsic coagulation mechanism inhibit blood platelet aggregation, in activator simultaneously be added salt,
The ingredients such as protective agent can play the function of ensureing blood coagulation.Thrombelastogram is carried out using the activator and activates hemostasis examination, it can
Accurately to measure fibrinogen content in blood sample.In addition, the prior art can effectively be solved in measurement fibrinogen
It is influenced when content by disturbing factors such as bubbles in sample optical exception, the finish of test cup, sample-adding, solves accuracy
The disadvantages of difference, poor repeatability, poor stability.
It should be noted that term " intrinsic coagulation activator " used in the present invention refers to that endogenous can be started
Ingredient of coagulation factor blood coagulation activity, such as Kaolin clay, ellagic acid etc.." extrinsic coagulation activator " refers to start outer
The ingredient of source property blood coagulation activity, such as tissue factor.
According to an embodiment of the invention, extrinsic coagulation activator is selected from tissue factor.Thus, it is possible to which effectively starting is solidifying
Blood access, to accurately determine fibrinogen content in blood sample based on clot strength MA.
It should be noted that the present invention does not make considered critical for the source of tissue factor, either buy, grant,
It is also possible to voluntarily prepare, it specifically can flexible choice according to the actual situation.According to a particular embodiment of the invention, tissue factor
Purchased from titanium source Bo Aote Bioisystech Co., Ltd.Thus, it is possible to further accurately measure fibrinogen content.
According to an embodiment of the invention, the concentration of extrinsic coagulation activator is 0.1~1mg/mL, platelet suppressant drug
Concentration is 1~20mg/ml.Inventor obtains the concentration of the two by many experiments, thus, it is possible to start extrinsic coagulation simultaneously
Access and inhibition platelet aggregation, so that accurate quantitative analysis detects fibrinogen content.
According to an embodiment of the invention, the platelet suppressant drug is in Yi Feiba object, tirofiban and Abciximab
At least one of.Inhibit the substance of platelet aggregation many currently, can play, still, inventors have found that and not all suppression
Preparation is suitable for detecting fibrinogen, for example, Cucumber while inhibiting platelet aggregation, can also inhibit exogenous
The effect of Coagulation pathways leads to not Accurate Determining fibrinogen content；Or the fibrin clot formation time is not suitable for.
Therefore, inventor has found by many experiments, these three substances of Yi Feiba object, tirofiban and Abciximab can effectively press down
Platelet aggregation processed, and extrinsic coagulation access is not influenced, the fibrin clot formation time is suitable for accurately measuring
Fibrinogen content in blood sample.
According to a particular embodiment of the invention, platelet suppressant drug is selected from Yi Feiba object, tirofiban, the mass ratio of the two
For 1:1.It is verified preferable for Platelet aggregation inhibitor effect using both platelet suppressant drugs.
According to an embodiment of the invention, protective agent in bovine serum albumin(BSA), casein and human serum albumins extremely
It is one of few.Protein component, which is added, has certain buffer capacity using protein as protective agent, while being capable of providing one kind
Good stability environment, to protect the other compositions in extrinsic coagulation activator not oxidized or group is caused to inactivate.
According to an embodiment of the invention, protectant concentration is 1~20mg/mL.Thus, it is possible to effectively guarantee blood coagulation
Function, it is ensured that the accuracy of testing result.
According to an embodiment of the invention, including buffer and salinity in buffer salt.Suitable ring is provided as a result, for blood coagulation
Border can effectively guarantee the function of blood coagulation, it is ensured that the accuracy of testing result.
According to an embodiment of the invention, the content of salinity is 0.1~1 mass %.Suitable ring is provided as a result, for blood coagulation
Border can effectively guarantee the function of blood coagulation, it is ensured that the accuracy of testing result.
According to an embodiment of the invention, the concentration of buffer is 10~100mmol/L, pH value is 6~8.It is as a result, blood coagulation
Suitable environment is provided, can effectively guarantee the function of blood coagulation, it is ensured that the accuracy of testing result.
According to an embodiment of the invention, salt is selected from least one of sodium chloride, potassium chloride and calcium chloride.It is as a result,
Blood coagulation provides suitable environment, can effectively guarantee the function of blood coagulation, it is ensured that the accuracy of testing result.
According to an embodiment of the invention, buffer is selected from least one of Tris, HEPES, PBS, glycine and MOPS.
Suitable environment is provided as a result, for blood coagulation, can effectively guarantee the function of blood coagulation, it is ensured that the accuracy of testing result.
According to an embodiment of the invention, functional fiber albumen activator further comprises preservative.Thus, it is possible to mention
High functional fiber albumen activator stability, extends the shelf life.
According to an embodiment of the invention, preservative is in Proclin300, potassium sorbate and methyl p-hydroxybenzoate
At least one of.Thus, it is possible to improve functional fiber albumen activator stability, extend the shelf life.
According to an embodiment of the invention, the concentration of preservative is 0.01~0.1 mass %.Thus, it is possible to improve functionality
Fibrinogen activator stability, extends the shelf life.
The method for preparing functional fiber albumen activator
In still another aspect of the invention, functional fiber albumen activator noted earlier is prepared the invention proposes a kind of
Method.According to an embodiment of the invention, this method comprises: preparing buffer；The salinity, protection are added into buffer
Agent stirs evenly, to obtain the first mixed liquor；Extrinsic coagulation activator is added into the first mixed liquor, stirs evenly, with
Just the second mixed liquor is obtained；Platelet suppressant drug is added into two mixed liquors of institute, stirs evenly, to obtain third mixed liquor；
And preservative is added into third mixed liquor, it stirs evenly, to obtain fibrinogen activator.Inventor is by a large amount of
The order of addition for testing each component obtained, the reagent stability made through preparation sequence, repeatability, accuracy have good
Good performance.The functional fiber albumen activator obtained as a result, can give full play to extrinsic coagulation effect and inhibit blood
Platelet aggtegation, the clot strength MA value and fibrinogen content of formation are proportional, to accurately measure blood
The content of fibrinogen in sample.
According to a particular embodiment of the invention, this method comprises the following steps:
S1 prepares the buffer solution of 10~100mmol/L, adjusts pH value to 6-8；
0.1~1 mass % salt is sequentially added in S2, Xiang Shangshu S1,1~20mg/ml protective agent stirs evenly；
1~10mg/ml platelet suppressant drug is added according to luxuriant and rich with fragrance Bart and tirofiban in S3 in above-mentioned S2；
0.1~1mg/ml extrinsic coagulation activator tissue factor is added in above-mentioned S3, stirs evenly by S4；
0.01~0.1 mass % preservative is added in above-mentioned S4, is dispensed and is lyophilized after mixing evenly by S5,
It is stored in 2~8 DEG C of environment.
It will be appreciated to those of skill in the art that above for feature described in functional fiber albumen activator
And advantage, it is equally applicable to the method for preparing functional fiber albumen activator, details are not described herein.
Kit
In still another aspect of the invention, the invention proposes a kind of kits.According to an embodiment of the invention, the kit
Including functional fiber albumen activator described above.It as a result, can be with using kit according to an embodiment of the present invention
Accurately measure fibrinogen content in blood sample.
According to an embodiment of the invention, kit further comprises calcium chloride solution, chlorination calcon is also known as coagulation factor
IV is the essential component for activating blood clotting.
According to an embodiment of the invention, the concentration of calcium chloride solution is 0.1~1mol/L.Inventor obtains by many experiments
To calcium chloride concentration, if calcium chloride concentration is too low, blood coagulation is too slow, and R value is too long；If calcium chloride concentration is excessively high, react too fast
Or causing ionic strength excessive, blood does not solidify.
According to an embodiment of the invention, kit further comprises specimen cup.Thus, it is possible to be used to load test sample,
Facilitate detection.
According to an embodiment of the invention, kit is used for thrombelastogram instrument hemostasis examination.By utilizing fibrinogen
Activator exogenous cruor pathway inhibits platelet aggregation simultaneously, rapidly and accurately detects in blood on thrombelastogram instrument
The clot strength of only fibrinogen contribution calculates fibrinogen content again, and it is stable, repeated which has performance
Good, long shelf-life, simple operation and other advantages.
It will be appreciated to those of skill in the art that above for feature described in functional fiber albumen activator
And advantage, it is equally applicable to the kit, details are not described herein.
The solution of the present invention is explained below in conjunction with embodiment.It will be understood to those of skill in the art that following
Embodiment is merely to illustrate the present invention, and should not be taken as limiting the scope of the invention.Particular technique or item are not specified in embodiment
Part, it described technology or conditions or is carried out according to the literature in the art according to product description.Agents useful for same or instrument
Production firm person is not specified in device, and being can be with conventional products that are commercially available.
Reagent:
Liquid bio preservative (Proclin300), content 98%, producer: Sigma company；
Bovine serum albumin(BSA), producer: ProliantBiologicals, Inc；
Tissue factor, producer: Boaote Biological Technology Co., Ltd., Taiyuan
Platelet suppressant drug: according to luxuriant and rich with fragrance Bart, producer: TRC；Tirofiban, producer: Aladdin reagent.
Embodiment 1
It present embodiments provides according to preparation coagulation activation agent, including following each group as following formula:
Fibrinogen activator is prepared in experimental group 1 in accordance with the following steps:
Step (1): disodium hydrogen phosphate 5g, sodium dihydrogen phosphate 0.6g, sodium chloride 8g, chlorination the preparation of PBS buffer solution: are weighed
1000mL purified water is added in potassium 0.2g, and stirring dissolves it sufficiently, adjusts pH value to 7.0；
Step (2): 20g bovine serum albumin(BSA) is added in the buffer of step (1)；
Step (3): being added eptifibatide 10g, tirofiban 10g in the solution of step (2), and stirring keeps it sufficiently mixed
Even, adjusting pH is 7.0-7.4；
Step (4): being added 1.0g tissue factor in the solution of step (3), and stirring dissolves it sufficiently, adjusts pH and is
7.0-7.4；
Step (5): being added Proclin300 1.0g and 0.5g gentamicin in the solution in step (4), and stirring makes it
Sufficiently dissolution, is made fibrinogen activator.Wherein gentamicin is primarily used to prevent the pollution of miscellaneous bacteria.
The fibrinogen activator being prepared is distributed into 10 μ L/ branch specimen cups and carries out freeze-drying preservation.
Fibrinogen activator is prepared in experimental group 2 in accordance with the following steps:
Step (1): trishydroxymethylaminomethane 2.4g, sodium chloride 8g, potassium chloride the preparation of Tris buffer: are weighed
1000ml purified water is added in 0.2g, and stirring dissolves it sufficiently, adjusts pH value to 7.0；
Step (2): being added 15g bovine serum albumin(BSA) in the buffer of step (1), and stirring dissolves it sufficiently；
Step (3): being added eptifibatide 10g, tirofiban 10g in the solution of step (2), and stirring keeps it sufficiently mixed
Even, adjusting pH is 7.0-7.4；
Step (4): being added 1.5g tissue factor in the solution of step (3), and stirring dissolves it sufficiently, adjusts pH and is
7.0-7.4；
Step (5): being added Proclin300 1.0g and 0.5g gentamicin in the solution in step (4), and stirring makes it
Sufficiently dissolution, is made fibrinogen activator.
The fibrinogen activator being prepared is distributed into 10 μ L/ branch specimen cups and carries out freeze-drying preservation.
Fibrinogen activator is prepared in experimental group 3 in accordance with the following steps:
Step (1): 4- hydroxyethyl piperazineethanesulfonic acid 4.7g, sodium chloride 7g, potassium chloride the preparation of HEPES buffer solution: are weighed
1000ml purified water is added in 0.5g, and stirring dissolves it sufficiently, adjusts pH value to 8.0；
Step (2): being added 10g bovine serum albumin(BSA) in the buffer of step (1),；
Step (3): being added eptifibatide 10g, tirofiban 10g in the solution of step (2), and stirring keeps it sufficiently mixed
Even, adjusting pH is 7.0-7.4；
Step (4): being added 0.5g tissue factor in the solution of step (3), and stirring dissolves it sufficiently, adjusts pH and is
7.0-7.4；
Step (5): being added Proclin300 1.0g and 0.5g gentamicin in the solution in step (4), and stirring makes it
Sufficiently dissolution, is made fibrinogen activator.
The fibrinogen activator being prepared is distributed into 10 μ L/ branch specimen cups and carries out freeze-drying preservation.
Inventor has found during the experiment simultaneously, if not containing protective agent in prepared activator, i.e., for example
Without containing bovine serum albumin(BSA), long term stability tests, the function of fibrinogen activator are carried out after placing for a long time
It substantially reduces.
Embodiment 2
Kit is obtained as follows, which includes fibrinogen activator, calcium chloride solution and sample
Product cup.
Wherein, fibrinogen activator is respectively the fibrinogen activator that experimental group 1-3 is prepared.
The concentration of calcium chloride solution is 0.2M: weighing 22.2g calcium chloride and is added in 1000mL physiological saline, stirring makes it
Sufficiently dissolution, is made into the calcium chloride solution of final concentration of 0.2M, is then distributed into 1mL/ branch.
Specimen cup is empty container, for loading detection sample and detection reagent, including blood sample, fibrinogen
Activator and calcium chloride solution.
Respectively according to as above, the first kit, the second kit, third kit are obtained.
Then 4 DEG C of refrigerations of kit are placed.
The precision of 3 functional fiber proteinogen detection reagent of embodiment is tested
It takes three kits of the invention to distinguish retest same sample 10 times, calculates in the horizontal present invention of different blood coagulations
Test precision.
For the kit that embodiment 2 is prepared, using following flow testing:
S1. by the reagent room temperature of refrigeration, restore to room temperature.
S2. the corresponding software of UD-T5000, input test sample information are opened, and selects " CFF " in test-types.
S3. specimen cup is loaded on thrombelastogram instrument TCH test channel.
S4. 20 μ L calcium chloride solutions are pipetted to be added to specimen cup bottom of a cup.
S5. 500 μ L sodium citrate anticoagulated whole bloods are pipetted to be added in the reagent bottle of fibrinogen activator, and use liquid relief
Rifle suction is made a call to 3 times.
S6. 340 μ L are pipetted to be mixed in the anticoagulated whole blood addition specimen cup of fibrinogen activator.
S7. specimen cup is pushed into test position, reference test bar is allocated to test position, and clicks START button on software and carry out
Test.
The testing result of sample such as table 1, wherein R (min) refers to that blood is put into thrombelastogram instrument and starts to first piece
Time (min) needed for fibrin clot forms (graphy figure amplitude reaches 2mm), MA (mm) are clot strength.
The precision test result of 1 functional fiber proteinogen detection reagent of table
From table 1, it can be seen that the CV of R, MA value of three kits of the invention illustrates the present invention within 10%
It is good that fibrinogen test kit detects precision.
The accelerated stability of 4 functional fiber proteinogen detection reagent of embodiment is tested
For the kit one that embodiment 2 is prepared, accelerates 0,2,4,6,8,10 day in 37 DEG C of insulating boxs respectively, take
Reagent after same sample accelerates carries out thrombelastogram instrument test.
S1. by the reagent room temperature of refrigeration, restore to room temperature.
S2. the corresponding software of UD-T5000, input test sample information are opened, and selects " CFF " in test-types.
S3. specimen cup is loaded on thrombelastogram instrument TCH test channel.
S4. 20 μ L calcium chloride solutions are pipetted, specimen cup bottom of a cup is added.
S5. 500 μ L sodium citrate anticoagulated whole bloods are pipetted to be added in the reagent bottle of fibrinogen activator, and use liquid relief
Rifle suction is made a call to 3 times.
S6. 340 μ L are pipetted to be mixed in the anticoagulated whole blood addition specimen cup of fibrinogen activator.
S7. specimen cup is pushed into test position, reference test bar is allocated to test position, and clicks START button on software and carry out
Test.
By taking the first, second kit as an example, the testing result of sample such as table 2.
The accelerated stability test result of 2 functional fiber proteinogen detection reagent of table
From table 2, it can be seen that the test R value of reagent of the present invention, MA value are as a result relatively very poor all to exist after accelerating 10 days
Within 10%, illustrate that functional fiber proteinogen detection kit accelerated stability of the present invention is good.
5 functional fiber proteinogen detection reagent long term stability tests of embodiment.
For the kit that embodiment 2 is prepared, 0,3,6,9,12,15,18 are stored under 2-8 DEG C of cold storage environment respectively
It a month, takes with a collection of Quality Control level 1 and Quality Control level 2, carries out thrombelastogram test with the reagent of experiment.
Thrombelastogram test is carried out with the reagent of experiment.It is specific as follows to measure process:
S1. the corresponding software of UD-T5000, input test sample information are opened, and selects " CFF " in test-types.
S2. specimen cup is loaded on thrombelastogram instrument TCH test channel.
S3. 20 μ L calcium chloride solutions are pipetted, specimen cup bottom of a cup is added.
S4. 500 μ L sodium citrate anticoagulated whole bloods are pipetted to be added in the reagent bottle of fibrinogen activator, and use liquid relief
Rifle suction is made a call to 3 times.
S5. 340 μ L are pipetted to be mixed in the anticoagulated whole blood addition specimen cup of fibrinogen activator.
S6. specimen cup is pushed into test position, reference test bar is allocated to test position, and clicks START button on software and carry out
Test.
By taking first dose of box as an example, the testing result of sample such as table 3.
3 functional fiber proteinogen extended storage stability test result of table
From table 3, it can be seen that test result after test R value, the MA value of reagent of the present invention store 18 months at 4 DEG C
It is relatively very poor all within 10%, illustrate that the extended storage stability of functional fiber proteinogen detection reagent of the present invention can be good
It is good.
6 functional fiber proteinogen detection reagent of embodiment detects contrast test with the FIB in blood clotting 4.
10 parts of fresh whole blood samples for choosing different fibrinogen contents, are prepared into the embodiment of the present invention 1,2 respectively
To the first kit and listed Siemens Healthcare Diagnostics Products GmbH production fibre
Fibrillarin original measurement reagent (freezing method) is simultaneously measured sample, acquired results is compared and correlation analysis, R2
Value is 0.9749, it can thus be appreciated that two methods test result correlation is without significant difference (Fig. 1).
The reagent of the present invention of table 4 with blood clotting 4 in contrast agent box testing result
In the description of this specification, reference term " one embodiment ", " some embodiments ", " example ", " specifically show
The description of example " or " some examples " etc. means specific features, structure, material or spy described in conjunction with this embodiment or example
Point is included at least one embodiment or example of the invention.In the present specification, schematic expression of the above terms are not
It must be directed to identical embodiment or example.Moreover, particular features, structures, materials, or characteristics described can be in office
It can be combined in any suitable manner in one or more embodiment or examples.In addition, without conflicting with each other, the skill of this field
Art personnel can tie the feature of different embodiments or examples described in this specification and different embodiments or examples
It closes and combines.
Although the embodiments of the present invention has been shown and described above, it is to be understood that above-described embodiment is example
Property, it is not considered as limiting the invention, those skilled in the art within the scope of the invention can be to above-mentioned
Embodiment is changed, modifies, replacement and variant.
Claims (10)
1. a kind of functional fiber albumen activator, which is characterized in that the functional fiber albumen activator includes:
Extrinsic coagulation activator, platelet suppressant drug, buffer salt and protective agent.
2. functional fiber albumen activator according to claim 1, which is characterized in that the extrinsic coagulation activation
Agent is selected from tissue factor；
Optionally, the concentration of the extrinsic coagulation activator is 0.1~1mg/mL.
3. functional fiber albumen activator according to claim 1, which is characterized in that the platelet suppressant drug choosing
From at least one of Yi Feiba object, tirofiban and Abciximab；
Optionally, the concentration of the platelet suppressant drug is 1~20mg/ml.
4. functional fiber albumen activator according to claim 1, which is characterized in that the protective agent is selected from ox blood
At least one of pure albumen, casein and human serum albumins；
Optionally, protectant concentration is 1~20mg/mL.
5. functional fiber albumen activator according to claim 1, which is characterized in that include slow in the buffer salt
Fliud flushing and salinity；
Optionally, the content of the salinity is 0.1~1 mass %；
Optionally, the concentration of the buffer is 10~100mmol/L, and pH value is 6~8.
6. functional fiber albumen activator according to claim 1, which is characterized in that the salt is selected from chlorination
At least one of sodium, potassium chloride and calcium chloride；
Optionally, the buffer is selected from least one of Tris, HEPES, PBS, glycine and MOPS.
7. functional fiber albumen activator according to claim 1, which is characterized in that the functional fiber albumen
Activator further comprises preservative；
Optionally, the preservative is selected from least one of Proclin300, potassium sorbate and methyl p-hydroxybenzoate；
Optionally, the concentration of the preservative is 0.01~0.1 mass %.
8. a kind of method for preparing any one of claim 1~7 functional fiber albumen activator, which is characterized in that
Include:
Prepare the buffer；
The salinity, protective agent are added into the buffer, stirs evenly, to obtain the first mixed liquor；
The extrinsic coagulation activator is added into first mixed liquor, stirs evenly, to obtain the second mixed liquor；
The platelet suppressant drug is added into second mixed liquor, stirs evenly, to obtain third mixed liquor；And
The preservative is added into the third mixed liquor, stirs evenly, to obtain the fibrinogen activator.
9. a kind of kit, which is characterized in that the kit includes the described in any item functional fibers of claim 1~7
Albumen activator；
Optionally, the kit further comprises calcium chloride solution；
Optionally, the concentration of the calcium chloride solution is 0.1~1mol/L.
10. kit according to claim 9, which is characterized in that the kit further comprises specimen cup；
Optionally, the kit is used for thrombelastogram instrument hemostasis examination.
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