CN117269507A - Preparation method of fibrinogen detection kit - Google Patents

Preparation method of fibrinogen detection kit Download PDF

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Publication number
CN117269507A
CN117269507A CN202311205488.7A CN202311205488A CN117269507A CN 117269507 A CN117269507 A CN 117269507A CN 202311205488 A CN202311205488 A CN 202311205488A CN 117269507 A CN117269507 A CN 117269507A
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fibrinogen
volume concentration
fibrinogen detection
mass volume
preset
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杨建鑫
李夏
张娟丽
李伟甲
赵晓培
杨会
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Zhengzhou Puwan Medical Technology Co ltd
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Zhengzhou Puwan Medical Technology Co ltd
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    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/5306—Improving reaction conditions, e.g. reduction of non-specific binding, promotion of specific binding
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
    • G01N2333/435—Assays involving biological materials from specific organisms or of a specific nature from animals; from humans
    • G01N2333/745—Assays involving non-enzymic blood coagulation factors
    • G01N2333/75—Fibrin; Fibrinogen

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  • Urology & Nephrology (AREA)
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  • General Health & Medical Sciences (AREA)
  • General Physics & Mathematics (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Chemical Kinetics & Catalysis (AREA)
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Abstract

The embodiment of the disclosure discloses a preparation method of a fibrinogen detection kit. One embodiment of the method comprises the following steps: dissolving trehalose, bovine serum albumin, arginine and urea by purified water; adding tirofiban, sodium salicylate, dipyridamole and tissue factor into the freeze-drying protective agent solution, and uniformly mixing; subpackaging the obtained test cup coating agent mother liquor into empty test cup; performing vacuum freeze drying treatment on the mother solution of the coating agent of the test cup after the split charging treatment to obtain a fibrinogen detection reagent; dissolving and uniformly mixing calcium chloride to obtain a calcium chloride solution as an activator; the fibrinogen detection reagent obtained and the activator obtained are combined into a fibrinogen detection kit. According to the embodiment, the practicability of the fibrinogen detection kit is improved, the operation steps when the fibrinogen detection kit is used are reduced, and the time consumption when the fibrinogen detection kit is used is shortened.

Description

Preparation method of fibrinogen detection kit
Technical Field
The embodiment of the disclosure relates to the technical field of medical instruments, in particular to a preparation method of a fibrinogen detection kit.
Background
The prepared fibrinogen detection kit can be used for determining the concentration value of blood and the like. Currently, in the preparation of fibrinogen detection reagents, the following methods are generally adopted: in the preparation process, a plurality of people of fibrinogen activators are freeze-dried in a penicillin bottle, wherein the fibrinogen activators are main components in a fibrinogen detection kit.
However, the inventors have found that when the fibrinogen detection kit is prepared in the above manner, there are often the following technical problems:
firstly, when the fibrinogen detection kit is prepared, a fibrinogen activator is freeze-dried in a penicillin bottle, when the fibrinogen activator in the penicillin bottle is used for detecting a blood sample, distilled water is required to be used for redissolving, and the taken blood sample is put into the penicillin bottle for activation, so that the operation steps of the fibrinogen detection kit using the fibrinogen activator freeze-dried in the penicillin bottle are more, and an operator needs to sample for multiple times, so that the prepared fibrinogen detection kit is poor in practicability, poor in stability of multiple times of sampling, less in times of consistent in the obtained sample amount, lower in accuracy of the prepared fibrinogen detection kit, and longer in time consumption of using the fibrinogen detection kit.
Secondly, when the fibrinogen detection kit is prepared, repeated tests are only carried out on each concentration value of liquid in the fibrinogen detection kit, so that the comprehensiveness of the tests on the detection kit is low, the accuracy of the prepared detection kit is low, and the prepared detection kit is poor in practicality.
Thirdly, freeze-drying the fibrinogen activator in a penicillin bottle, wherein the dose of the fibrinogen activator freeze-dried in the penicillin bottle is a multi-person dose, and the opened detection reagent needs to be used up once, so that the use rate of the detection reagent is low, and the waste of the detection reagent is caused.
The above information disclosed in this background section is only for enhancement of understanding of the background of the inventive concept and, therefore, may contain information that does not form the prior art that is already known to those of ordinary skill in the art in this country.
Disclosure of Invention
The disclosure is in part intended to introduce concepts in a simplified form that are further described below in the detailed description. The disclosure is not intended to identify key features or essential features of the claimed subject matter, nor is it intended to be used to limit the scope of the claimed subject matter.
Some embodiments of the present disclosure propose methods of preparing fibrinogen detection kits to address one or more of the technical problems mentioned in the background section above.
In a first aspect, some embodiments of the present disclosure provide a method of preparing a fibrinogen detection kit, the method comprising: dissolving trehalose, bovine serum albumin, arginine and urea by purified water to obtain a freeze-drying protective agent solution; adding tirofiban, sodium salicylate, dipyridamole and tissue factor into the obtained freeze-drying protective agent solution, and uniformly mixing to obtain test cup coating agent mother liquor; subpackaging the obtained test cup coating agent mother liquor to each test cup empty cup so as to perform subpackaging treatment on the obtained test cup coating agent mother liquor; performing vacuum freeze drying treatment on the packaged test cup coating agent mother liquor to obtain a fibrinogen detection reagent, wherein the obtained fibrinogen detection reagent is coated on the test cup; dissolving and uniformly mixing calcium chloride to obtain a calcium chloride solution as an activator; the fibrinogen detection reagent obtained and the activator obtained are combined into a fibrinogen detection kit.
The above embodiments of the present disclosure have the following advantageous effects: by the preparation method of the fibrinogen detection kit, the practicability and the accuracy of the prepared fibrinogen detection kit are improved, the sampling times of operators are reduced, the steps of using the fibrinogen detection kit are simplified, and the time consumption of using the fibrinogen detection kit is shortened. Specifically, the prepared fibrinogen detection kit is poor in practicality, and the time consumed for using the prepared fibrinogen detection kit is long because: when the fibrinogen detection kit is prepared, the fibrinogen activator is freeze-dried in a penicillin bottle, distilled water is required to be used for redissolving and the taken blood sample is required to be put in the penicillin bottle for activation when the fibrinogen activator in the penicillin bottle is used for detecting the blood sample, so that the operation steps of the fibrinogen detection kit using the fibrinogen activator freeze-dried in the penicillin bottle are more, and an operator needs to sample for many times, so that the prepared fibrinogen detection kit is poor in practicability, poor in stability of sampling for many times, less in the number of times of consistent sample amount, lower in accuracy of the prepared fibrinogen detection kit and longer in time consumption when the fibrinogen detection kit is used. In the preparation method of the fibrinogen detection kit of some embodiments of the present disclosure, first, trehalose, bovine serum albumin, arginine and urea are dissolved by purified water to obtain a lyoprotectant solution. Thus, a lyoprotectant solution can be obtained, which can be used for preparing fibrinogen detection reagents. And then, adding tirofiban, sodium salicylate, dipyridamole and tissue factor into the obtained freeze-drying protective agent solution, and uniformly mixing to obtain test cup coating agent mother liquor. Thus, a fibrinogen detection reagent mother solution can be obtained. And then, sub-packaging the obtained test cup coating agent mother liquor into empty test cup to sub-package the obtained test cup coating agent mother liquor. Thus, each test cup of the same dose can be obtained. And then, carrying out vacuum freeze drying treatment on the mother solution of the coating agent of the test cup after the split treatment to obtain the fibrinogen detection reagent, wherein the obtained fibrinogen detection reagent is coated on the test cup. Thus, a fibrinogen detection reagent coated on the test cup can be obtained. And then, carrying out water dissolution and uniform mixing treatment on the calcium chloride to obtain a calcium chloride solution serving as an activator. Thus, an activator solution can be obtained. Finally, the fibrinogen detection reagent and the activator are combined to form the fibrinogen detection kit. Thus, the prepared fibrinogen detection kit can be obtained. Also because when the fibrinogen detection kit is prepared, the fibrinogen detection reagent contained in the fibrinogen detection kit is not freeze-dried in the penicillin bottle, but is coated in the test cup, when the fibrinogen detection kit is used, distilled water is not required to be used for redissolution, and the taken blood sample is not required to be put into the penicillin bottle for activation, the steps of using the prepared fibrinogen detection kit are simplified, the practicability and the accuracy of using the fibrinogen detection kit are improved, and the time consumption of using the prepared fibrinogen detection kit is shortened.
Drawings
The above and other features, advantages, and aspects of embodiments of the present disclosure will become more apparent by reference to the following detailed description when taken in conjunction with the accompanying drawings. The same or similar reference numbers will be used throughout the drawings to refer to the same or like elements. It should be understood that the figures are schematic and that elements and components are not necessarily drawn to scale.
Fig. 1 is a flow chart of some embodiments of methods of making fibrinogen detection kits according to the present disclosure.
Detailed Description
Embodiments of the present disclosure will be described in more detail below with reference to the accompanying drawings. While certain embodiments of the present disclosure are shown in the drawings, it should be understood that the present disclosure may be embodied in various forms and should not be construed as limited to the embodiments set forth herein. Rather, these embodiments are provided so that this disclosure will be thorough and complete. It should be understood that the drawings and embodiments of the present disclosure are for illustration purposes only and are not intended to limit the scope of the present disclosure.
It should be noted that, for convenience of description, only the portions related to the present invention are shown in the drawings. Embodiments of the present disclosure and features of embodiments may be combined with each other without conflict.
It should be noted that the terms "first," "second," and the like in this disclosure are merely used to distinguish between different devices, modules, or units and are not used to define an order or interdependence of functions performed by the devices, modules, or units.
It should be noted that references to "one", "a plurality" and "a plurality" in this disclosure are intended to be illustrative rather than limiting, and those of ordinary skill in the art will appreciate that "one or more" is intended to be understood as "one or more" unless the context clearly indicates otherwise.
The present disclosure will be described in detail below with reference to the accompanying drawings in conjunction with embodiments.
Fig. 1 illustrates a flow 100 of some embodiments of a method of making a fibrinogen detection kit according to the present disclosure. The preparation method of the fibrinogen detection kit comprises the following steps:
and 101, dissolving trehalose, bovine serum albumin, arginine and urea by purified water to obtain a freeze-drying protective agent solution.
In some embodiments, trehalose, bovine serum albumin, arginine, and urea may be dissolved by purified water to obtain a lyoprotectant solution. Wherein the mass volume concentration of the trehalose can be 0.6%. The mass volume concentration of the bovine serum albumin may be 0.2%. The arginine may be present in a mass volume concentration of 0.06%. The mass volume concentration of the urea may be 0.6%. In practice, purified water may be added to dissolve the trehalose, the bovine serum albumin, the arginine and the urea to obtain a lyoprotectant solution.
And 102, adding tirofiban, sodium salicylate, dipyridamole and tissue factor into the obtained freeze-drying protective agent solution, and uniformly mixing to obtain test cup coating agent mother liquor.
In some embodiments, tirofiban, sodium salicylate, dipyridamole and tissue factor may be added to the resulting lyoprotectant solution and subjected to a homogenization process to obtain a test cup coating agent mother liquor. Wherein, the mass volume concentration of the tirofiban can be 0.5 percent. The sodium salicylate may have a mass volume concentration of 4.0%. The mass volume concentration of the dipyridamole may be 0.5%. The volume concentration of the tissue factor may be 0.4. Mu.g/ml. In practice, the lyoprotectant solution added with the tirofiban, the sodium salicylate, the dipyridamole and the tissue factor can be mixed upside down to obtain the mother solution of the test cup coating agent.
And step 103, sub-packaging the obtained test cup coating agent mother liquor into empty test cup to perform sub-packaging treatment on the obtained test cup coating agent mother liquor.
In some embodiments, the resulting test cup coating agent mother liquor may be packaged into empty test cup to provide a packaged treatment of the resulting test cup coating agent mother liquor. Wherein the volume of the mother solution of the coating agent of the test cup divided in each empty test cup is 20 mu l.
And 104, performing vacuum freeze drying treatment on the separated test cup coating agent mother liquor to obtain the fibrinogen detection reagent.
In some embodiments, the test cup coating agent mother liquor after the split treatment can be subjected to vacuum freeze drying treatment to obtain the fibrinogen detection reagent. Wherein the resulting fibrinogen detection reagent is coated on the test cup.
And 105, carrying out water dissolution and uniform mixing treatment on the calcium chloride to obtain a calcium chloride solution serving as an activator.
In some embodiments, the calcium chloride may be subjected to a water-dissolving and homogenization treatment to provide a calcium chloride solution as an activator. Wherein the mass concentration of the calcium chloride is 0.2mol/L. In practice, first, purified water may be added to subject the above calcium chloride to water-soluble treatment. Then, the calcium chloride solution after water treatment can be mixed upside down to obtain the activator.
Step 106, combining the obtained fibrinogen detection reagent and the obtained activator into a fibrinogen detection kit.
In some embodiments, the resulting fibrinogen detection reagent and the resulting activator may be combined into a fibrinogen detection kit. Wherein the fibrinogen detection kit may comprise a fibrinogen detection reagent and the resulting activator.
The concentration of the tissue factor is the volume concentration, the concentration of the calcium chloride is the mass concentration of the substance, and the concentrations of the other components are all mass volume concentrations, and in practice, the concentrations are converted into specific grams according to the volume of the solution to be prepared. Adding purified water into the solution to enable the scale mark corresponding to the volume of the solution to reach a preset scale mark, wherein the preset scale mark can be the scale mark corresponding to the volume of the solution to be configured in practice. Ten human copies of the test reagent may be included in each fibrinogen test kit.
Alternatively, the low temperature, dry protectant included in the lyoprotectant solution described above may be prepared from one or more of a saccharide protectant and a polymeric protectant. Wherein, the saccharide protecting agent can be prepared from one or more of dextran, sucrose and trehalose. The polymer protective agent can be prepared from one or more of polyvinylpyrrolidone (PVP), bovine Serum Albumin (BSA), polyethylene glycol and dextran.
Alternatively, the bulking agent included in the lyoprotectant solution described above may be prepared from one or more of glycine, arginine, serine.
Alternatively, the stabilizer included in the lyoprotectant solution may be prepared from one or more of urea, metformin, ethylenediamine. Wherein, the freeze-drying protective agent solution can comprise low temperature, a drying protective agent, a filling agent and a stabilizing agent.
Alternatively, the platelet glycoprotein IIa/IIIa receptor inhibitor included in the test cup coating mother liquor may be prepared from one or more of tirofiban, acipimab and eptifibatide.
Optionally, thromboxane TXA contained in the mother solution of the test cup coating agent 2 The synthesis inhibitor can be prepared from one or more of aspirin, sodium salicylate and ibuprofen.
Alternatively, the cyclic adenosine monophosphate (cAMP) enhancer included in the test cup coating agent mother liquor described above may be dipyridamole. Wherein the test cup coating mother liquor may comprise platelet glycoprotein IIa/IIIa receptor inhibitor and thromboxane TXA 2 Synthetic inhibitors and cyclic adenosine monophosphate (cAMP) enhancers.
In some alternative implementations of some embodiments, the assay may be performed during the preparation of the fibrinogen detection kit by:
in the first step, each component included in the fibrinogen detection reagent in the fibrinogen detection kit is tested to obtain a concentration value of each component corresponding to the fibrinogen detection reagent. Wherein the component concentration value of the respective component concentration values may correspond to the component of the respective components. The above ingredients may include trehalose, bovine serum albumin, arginine, urea, tissue factor, tirofiban, sodium salicylate, and dipyridamole. The respective component concentration values may include a mass-volume concentration value of trehalose, a mass-volume concentration value of bovine serum albumin, a mass-volume concentration value of arginine, a mass-volume concentration value of urea, a mass-volume concentration value of tirofiban, a mass-volume concentration value of sodium salicylate, a mass-volume concentration value of dipyridamole, and a volume concentration value of tissue factor.
And secondly, performing validity test on the fibrinogen detection kit to determine whether the fibrinogen detection kit meets preset validity conditions, and performing precision test on the fibrinogen detection kit to determine whether the fibrinogen detection kit meets preset precision conditions.
In some alternative implementations of some embodiments, the fibrinogen detection reagent in the fibrinogen detection kit may be tested for each component included in the fibrinogen detection reagent by:
first, a first set of values corresponding to a first predetermined mass-volume concentration of trehalose included in the fibrinogen test reagent is determined. Wherein the first set of predetermined mass volume concentration values may include 0.3%, 0.6%, and 0.9%. The first set of preset mass volume concentration values may characterize preset individual concentration values of trehalose. In practice, first, a predetermined range of mass-volume concentration values corresponding to trehalose may be determined. Wherein, the preset mass volume concentration value range of the trehalose can be 0.1-1.0%. And then, selecting each preset mass volume concentration value from the preset mass volume concentration value range of the trehalose as a first preset mass volume concentration value set.
And a second step of determining a second set of predetermined mass-volume concentration values corresponding to bovine serum albumin included in the fibrinogen test reagent. Wherein the second set of predetermined mass volume concentration values may include 0.2%, 0.4%, and 0.6%. The second set of predetermined mass volume concentration values may characterize respective concentration values of predetermined bovine serum albumin. In practice, first, a predetermined range of mass-volume concentration values corresponding to bovine serum albumin may be determined. Wherein, the preset mass volume concentration value range of the bovine serum albumin can be 0.1-1.0%. Then, each preset mass-volume concentration value is selected from the preset mass-volume concentration value range of the bovine serum albumin as a second preset mass-volume concentration value set.
And thirdly, determining a third preset mass volume concentration value set corresponding to the arginine contained in the fibrinogen detection reagent. Wherein the third set of preset mass volume concentration values may include 0.03%, 0.06%, and 0.09%. The third set of preset mass volume concentration values may characterize preset individual concentration values of arginine. In practice, first, a predetermined range of mass volume concentration values corresponding to arginine may be determined. Wherein, the preset mass volume concentration value range of the arginine can be 0.01-0.1%. Then, each preset mass volume concentration value is selected from the preset mass volume concentration value range of the arginine as a third preset mass volume concentration value set.
And a fourth step of determining a fourth set of preset mass-volume concentration values corresponding to urea included in the fibrinogen detection reagent. Wherein the fourth set of predetermined mass volume concentration values may include 0.3%, 0.6%, and 0.9%. The fourth set of preset mass volume concentration values may represent respective concentration values of the preset urea. In practice, first, a range of preset mass-volume concentration values corresponding to urea may be determined. Wherein, the preset mass volume concentration value range of the urea can be 0.1-1.0%. And then, selecting each preset mass volume concentration value from the preset mass volume concentration value range of the urea as a fourth preset mass volume concentration value set.
And fifthly, determining a fifth preset volume concentration value set corresponding to the tissue factor included in the fibrinogen detection reagent. Wherein, the fifth set of preset volume concentration values may include 0.2, 0.4 and 0.6. The fifth set of preset volume concentration values may characterize respective concentration values of a preset tissue factor. In practice, first, a range of preset volume concentration values for the corresponding tissue factor may be determined. Wherein, the preset volume concentration value range of the tissue factor can be 0.1-1.0 mug/ml. Then, each preset volume concentration value is selected from the preset volume concentration value range of the tissue factor to be used as a fifth preset volume concentration value set.
And a sixth step of determining a sixth set of preset mass-volume concentration values corresponding to tirofiban included in the fibrinogen test reagent. Wherein the sixth set of preset mass volume concentration values may include 0.5%, 1.0%, and 1.5%. The sixth set of preset mass volume concentration values may characterize the respective concentration values of the preset tirofiban. In practice, first, a preset mass-volume concentration value range corresponding to tirofiban may be determined. Wherein, the preset mass volume concentration value range of the tirofiban can be 0.1-2.0%. Then, each preset mass-volume concentration value is selected from the preset mass-volume concentration value range of tirofiban as a sixth preset mass-volume concentration value set.
And a seventh step of determining a seventh set of preset mass-volume concentration values corresponding to sodium salicylate included in the fibrinogen detection reagent. Wherein, the seventh set of preset mass volume concentration values may include 2.0%, 3.0%, and 4.0%. The seventh set of preset mass volume concentration values may characterize the respective concentration values of the preset sodium salicylate. In practice, first, a predetermined range of mass volume concentration values corresponding to sodium salicylate may be determined. Wherein, the preset mass volume concentration value range of the sodium salicylate can be 1.0-5.0%. Then, each preset mass volume concentration value is selected from the preset mass volume concentration value range of the sodium salicylate to be used as a seventh preset mass volume concentration value set.
And an eighth step of determining an eighth set of values of a predetermined mass volume concentration corresponding to dipyridamole included in the fibrinogen test reagent. Wherein, the eighth set of preset mass volume concentration values may include 0.5%, 1.0% and 1.5%. The eighth set of preset mass volume concentration values may characterize the respective concentration values of preset dipyridamole. In practice, first, a range of preset mass-volume concentration values corresponding to dipyridamole may be determined. Wherein, the preset mass volume concentration value range of the dipyridamole can be 0.1-2.0%. Then, each preset mass volume concentration value is selected from the preset mass volume concentration value range of dipyridamole as an eighth preset mass volume concentration value set.
And a ninth step of combining each fibrinogen detection reagent having different component concentration values according to the first preset mass volume concentration value set, the second preset mass volume concentration value set, the third preset mass volume concentration value set, the fourth preset mass volume concentration value set, the fifth preset volume concentration value set, the sixth preset mass volume concentration value set, the seventh preset mass volume concentration value set, and the eighth preset mass volume concentration value set corresponding to the fibrinogen detection reagent. In practice, the first set of preset mass volume concentration values, the second set of preset mass volume concentration values, the third set of preset mass volume concentration values, the fourth set of preset mass volume concentration values, the fifth set of preset volume concentration values, the sixth set of preset mass volume concentration values, the seventh set of preset mass volume concentration values and each of the preset mass volume concentration values in the eighth set of preset mass volume concentration values may be combined by using an orthogonal combination method, so as to obtain each fibrinogen detection reagent having different component concentration values. For example, the fibrinogen detection reagents having different concentration values of the respective components may be as shown in the following table:
And tenth, subpackaging and freeze-drying each fibrinogen detection reagent in the fibrinogen detection reagents according to a preset dosage to obtain each fibrinogen detection reagent cup. For example, the predetermined dose may be 20 μl/cup. In practice, each fibrinogen test reagent may be dispensed and lyophilized at 20. Mu.l/cup.
Eleventh, adding the predetermined dose of the calcium chloride solution to each of the fibrinogen measurement reagent cups to obtain each fibrinogen measurement reagent cup containing the calcium chloride solution as each target fibrinogen measurement reagent cup. Wherein the mass concentration of the substance of the calcium chloride solution can be 0.2mol/L.
And a twelfth step of detecting the fibrinogen function of the target blood sample according to the target fibrinogen detection reagent cups to obtain detection results corresponding to the target fibrinogen detection reagent cups, wherein the target fibrinogen detection reagent cups in the target fibrinogen detection reagent cups correspond to the detection results in the detection results. The detection result in each detection result may be an average value of three detection results of the corresponding fibrinogen detection reagent cup of interest. For example, the respective test results corresponding to the respective target fibrinogen test reagent cups described above can be shown in the following table:
Thirteenth, determining the concentration value of each component corresponding to the detection result satisfying the preset result condition as the concentration value of each component corresponding to each component in the fibrinogen detection reagent. The preset result condition may be a detection result with the smallest average value among the detection results of the target fibrinogen detection reagent cups.
In some alternative implementations of some embodiments, the fibrinogen detection kit may be subjected to a validity test to determine whether the fibrinogen detection kit meets a preset validity condition, and a precision test to determine whether the fibrinogen detection kit meets a preset precision condition by:
in the first step, the fibrinogen function of each blood sample in the blood sample collection is detected by the detection reagent in the existing detection kit, so as to obtain the detection result of each target blood sample corresponding to the blood sample collection. The existing detection kit can be an imported kit suitable for detection of a thromboelastography instrument. The number of blood samples in the blood sample set may be a preset number. For example, the preset number may be 10.
And a second step of detecting the fibrinogen function of each blood sample in the blood sample set by using the fibrinogen detection reagent in the fibrinogen detection kit to obtain each blood sample detection result corresponding to the blood sample set, wherein the target blood sample detection result in each target blood sample detection result corresponds to the blood sample detection result in each blood sample detection result. For example, the detection results of the set of blood samples by the detection reagent in the conventional detection kit and the fibrinogen detection reagent in the fibrinogen detection kit are shown in the following table:
test sample MA/mm of the reagent to be detected Certain inlet reagent MA/mm
1 18.4 26.1
2 20.5 27.0
3 15.3 24.3
4 24.7 31.2
5 19.8 28.4
6 13.6 21.6
7 22.3 34.0
8 14.0 22.5
9 11.9 18.8
10 12.2 19.6
And a third step of determining that the fibrinogen detection reagent satisfies the preset validity condition in response to determining that each of the target blood sample detection results is greater than the corresponding blood sample detection result. The preset validity condition may be that each target blood sample detection result of the target blood sample detection results is larger than the corresponding blood sample detection result.
And step four, selecting each fibrinogen detection kit with preset numerical parts from the fibrinogen detection kit collection at will, wherein the batch numbers corresponding to the fibrinogen detection kits in the fibrinogen detection kit collection are consistent. For example, the preset number of parts may be 10 parts.
Fifthly, detecting the fibrinogen function of the target blood sample according to the selected fibrinogen detection kits to obtain detection results of the target blood samples corresponding to the target blood samples. In practice, the target blood sample may be tested using the thromboelastography apparatus described above.
And sixthly, determining a variation coefficient corresponding to each target blood sample detection result according to each target blood sample detection result. In practice, first, an average value of the detection results corresponding to each target blood sample may be determined. Then, standard deviations corresponding to the detection results of the respective target blood sample can be obtained from the average values. Finally, the ratio of the average value and the standard deviation can be determined as a variation coefficient corresponding to the detection result of each target blood sample. For example, the respective target blood sample detection results corresponding to the above-mentioned target blood sample, and the average value, standard deviation, and variation coefficient corresponding to the respective target blood sample detection results may be as shown in the following table:
number of times of detection Fibrinogen function MA/mm
1 15.3
2 14.2
3 13.6
4 15.0
5 15.3
6 14.8
7 13.6
8 14.0
9 13.2
10 12.7
Average value of 14.2
Standard deviation of 0.86
Coefficient of variation (%) 6.1
Seventh, determining that the fibrinogen detection kit satisfies the preset precision condition in response to determining that each of the variation coefficients is smaller than a preset variation coefficient. The predetermined coefficient of variation may be 15%.
The above technical solution is an invention point of the embodiments of the present disclosure, and solves the second technical problem mentioned in the background art, in which when the fibrinogen detection kit is prepared, only each concentration value of the liquid in the fibrinogen detection kit is tested repeatedly, resulting in lower comprehensiveness of testing the detection kit and lower accuracy of the prepared detection kit, which results in poor practicability of the prepared detection kit. The comprehensiveness of the test on the detection kit is poor, the detection accuracy of the prepared detection kit is low, and the factors of poor practicability of the prepared detection kit are often as follows: only the concentration values of the liquids in the test kit were tested repeatedly. If the factors are solved, the comprehensiveness of testing the detection kit can be improved, the detection accuracy of the prepared detection kit can be improved, and the practicability of the prepared detection kit can be improved. In order to achieve the effect, when the detection kit is detected, the concentration value in the detection kit is detected, and the precision and the effectiveness of the prepared detection kit are detected, so that the comprehensiveness of the detection kit detection is improved, the detection accuracy of the prepared detection kit is improved, and the practicability of the prepared detection kit is improved.
Optionally, after step 106, the target blood sample may also be tested according to the prepared fibrinogen test kit by:
in the first step, information of a target blood sample is input to an information interface to be tested applied by a thromboelastography machine, and a test type is selected as fibrinogen detection. The information of the target blood sample may be name information corresponding to the target blood sample. The application of the thrombelastogram instrument can be application software of the corresponding thrombelastogram instrument. The fibrinogen test may be a test for the fibrinogen function of the target blood sample. The thromboelastography application described above may be software associated with a thromboelastography.
In a second step, the test cup is loaded onto the channel of the thromboelastography machine. Wherein, fibrinogen detection reagent is coated on the test cup. The fibrinogen detection reagent may include trehalose, bovine serum albumin, arginine, urea, tissue factor, tirofiban, sodium salicylate, and dipyridamole. Wherein, the pH value range of the fibrinogen detection reagent can be 7.0-7.5.
And thirdly, placing the calcium chloride solution at the bottom of the test cup to obtain the test cup containing the calcium chloride solution. Wherein the volume of the calcium chloride solution placed in the bottom of the test cup may be 20. Mu.l. In practice, 20. Mu.l of calcium chloride solution may be placed in the bottom of the test cup.
And fourthly, transferring the anticoagulated whole blood sample of sodium citrate corresponding to the target blood sample to a test cup containing a calcium chloride solution, and uniformly mixing. In practice, first, a sample of anticoagulated whole blood of sodium citrate corresponding to the target blood sample may be transferred to a test cup containing a calcium chloride solution. Then, the test cup containing the calcium chloride solution after the sodium citrate anticoagulation whole blood sample is removed can be mixed up and down reversely, so that the test cup containing the calcium chloride solution is uniformly mixed.
And fifthly, placing the uniformly mixed test cup at the top end of a channel of the thromboelastography, and placing a test rod of the thromboelastography into a test position. Wherein, the test rod can be the component of the thromboelastography instrument.
And sixthly, selecting a fibrinogen detection channel in the application of the thromboelastography, and clicking to start detection so as to detect the fibrinogen function of the sodium citrate anticoagulated whole blood sample of the target blood sample. The fibrinogen detection channel may be a channel for detecting fibrinogen functions of the blood sample.
Alternatively, the sample of sodium citrate anticoagulated whole blood corresponding to the target blood sample may be removed to a test cup containing a calcium chloride solution by the following steps:
Firstly, a preset volume of sodium citrate anticoagulated whole blood sample can be removed from the sodium citrate anticoagulated whole blood sample of the target blood sample to a test cup containing the calcium chloride solution. For example, the preset volume may be 340 μl.
And secondly, uniformly mixing the calcium chloride solution and the sodium citrate anticoagulated whole blood sample in a test cup. In practice, the test cup containing the calcium chloride solution and the sodium citrate anticoagulated whole blood sample can be mixed up and down reversely to uniformly mix the solution in the test cup.
The above technical solution is an invention point of the embodiments of the present disclosure, and solves the third "the technical problem mentioned in the background art, in which the dose of the fibrinogen activator lyophilized in the penicillin bottle is a multiple dose, and the opened detection reagent needs to be used up once, which results in lower usage rate of the detection reagent and waste of the detection reagent". Factors that lead to lower usage of the detection reagent and waste of the detection reagent are often as follows: the reagent to be detected is a plurality of persons, not one person, so that when the blood sample is detected, the detection reagent is not used up and cannot be stored, thereby wasting the detection reagent. If the above factors are solved, waste of detection reagent can be reduced, and in order to achieve the effect, when the fibrinogen detection kit is prepared, one person of detection reagent is coated on the test cup, and each cup can only be used by one person. Therefore, the waste caused by the fact that a plurality of people do not use the detection reagent can be reduced.
The above embodiments of the present disclosure have the following advantageous effects: by the preparation method of the fibrinogen detection kit, the practicability and the accuracy of the prepared fibrinogen detection kit are improved, the sampling times of operators are reduced, the steps of using the fibrinogen detection kit are simplified, and the time consumption of using the fibrinogen detection kit is shortened. Specifically, the prepared fibrinogen detection kit is poor in practicality, and the time consumed for using the prepared fibrinogen detection kit is long because: when the fibrinogen detection kit is prepared, the fibrinogen activator is freeze-dried in a penicillin bottle, distilled water is required to be used for redissolving and the taken blood sample is required to be put in the penicillin bottle for activation when the fibrinogen activator in the penicillin bottle is used for detecting the blood sample, so that the operation steps of the fibrinogen detection kit using the fibrinogen activator freeze-dried in the penicillin bottle are more, and an operator needs to sample for many times, so that the prepared fibrinogen detection kit is poor in practicability, poor in stability of sampling for many times, less in the number of times of consistent sample amount, lower in accuracy of the prepared fibrinogen detection kit and longer in time consumption when the fibrinogen detection kit is used. In the preparation method of the fibrinogen detection kit of some embodiments of the present disclosure, first, trehalose, bovine serum albumin, arginine and urea are dissolved by purified water to obtain a lyoprotectant solution. Thus, a lyoprotectant solution can be obtained, which can be used for preparing fibrinogen detection reagents. And then, adding tirofiban, sodium salicylate, dipyridamole and tissue factor into the obtained freeze-drying protective agent solution, and uniformly mixing to obtain test cup coating agent mother liquor. Thus, a fibrinogen detection reagent mother solution can be obtained. And then, sub-packaging the obtained test cup coating agent mother liquor into empty test cup to sub-package the obtained test cup coating agent mother liquor. Thus, each test cup of the same dose can be obtained. And then, carrying out vacuum freeze drying treatment on the mother solution of the coating agent of the test cup after the split treatment to obtain the fibrinogen detection reagent, wherein the obtained fibrinogen detection reagent is coated on the test cup. Thus, a fibrinogen detection reagent coated on the test cup can be obtained. And then, carrying out water dissolution and uniform mixing treatment on the calcium chloride to obtain a calcium chloride solution serving as an activator. Thus, an activator solution can be obtained. Finally, the fibrinogen detection reagent and the activator are combined to form the fibrinogen detection kit. Thus, the prepared fibrinogen detection kit can be obtained. Also because when the fibrinogen detection kit is prepared, the fibrinogen detection reagent contained in the fibrinogen detection kit is not freeze-dried in the penicillin bottle, but is coated in the test cup, when the fibrinogen detection kit is used, distilled water is not required to be used for redissolution, and the taken blood sample is not required to be put into the penicillin bottle for activation, the steps of using the prepared fibrinogen detection kit are simplified, the practicability and the accuracy of using the fibrinogen detection kit are improved, and the time consumption of using the prepared fibrinogen detection kit is shortened.
The foregoing description is only of the preferred embodiments of the present disclosure and description of the principles of the technology being employed. It will be appreciated by those skilled in the art that the scope of the invention in the embodiments of the present disclosure is not limited to the specific combination of the above technical features, but encompasses other technical features formed by any combination of the above technical features or their equivalents without departing from the spirit of the invention. Such as the above-described features, are mutually substituted with (but not limited to) the features having similar functions disclosed in the embodiments of the present disclosure.

Claims (6)

1. A method for preparing a fibrinogen assay kit comprising:
dissolving trehalose, bovine serum albumin, arginine and urea by purified water to obtain a freeze-drying protective agent solution;
adding tirofiban, sodium salicylate, dipyridamole and tissue factor into the obtained freeze-drying protective agent solution, and uniformly mixing to obtain test cup coating agent mother liquor;
subpackaging the obtained test cup coating agent mother liquor to each test cup empty cup so as to perform subpackaging treatment on the obtained test cup coating agent mother liquor;
Performing vacuum freeze drying treatment on the packaged test cup coating agent mother liquor to obtain a fibrinogen detection reagent, wherein the obtained fibrinogen detection reagent is coated on the test cup;
dissolving and uniformly mixing calcium chloride to obtain a calcium chloride solution as an activator;
the fibrinogen detection reagent obtained and the activator obtained are combined into a fibrinogen detection kit.
2. The method of claim 1, wherein the trehalose is present at a mass volume concentration of 0.6%, the bovine serum albumin is present at a mass volume concentration of 0.2%, the arginine is present at a mass volume concentration of 0.06%, the urea is present at a mass volume concentration of 0.6%, and the calcium chloride is present at a mass concentration of 0.2mol/L.
3. The method of claim 1, wherein the tirofiban has a mass volume concentration of 0.5%, the sodium salicylate has a mass volume concentration of 4.0%, the dipyridamole has a mass volume concentration of 0.5%, and the tissue factor has a volume concentration of 0.4 μg/ml.
4. The method according to claim 1, wherein the fibrinogen detection reagent has a ph in the range of 7.0 to 7.5.
5. The method of claim 1, wherein the fibrinogen detection kit is prepared by the following test steps:
testing each component included in the fibrinogen detection reagent in the fibrinogen detection kit to obtain each component concentration value corresponding to the fibrinogen detection reagent, wherein each component concentration value in each component concentration value corresponds to a component in each component, and each component concentration value comprises a mass volume concentration value of trehalose, a mass volume concentration value of bovine serum albumin, a mass volume concentration value of arginine, a mass volume concentration value of urea, a mass volume concentration value of tirofiban, a mass volume concentration value of sodium salicylate, a mass volume concentration value of dipyridamole and a volume concentration value of tissue factor;
and performing a validity test on the fibrinogen detection kit to determine whether the fibrinogen detection kit meets a preset validity condition, and performing a precision test on the fibrinogen detection kit to determine whether the fibrinogen detection kit meets the preset precision condition.
6. The method according to claim 5, wherein the testing the respective components included in the fibrinogen detection reagent in the fibrinogen detection kit to obtain respective component concentration values corresponding to the fibrinogen detection reagent comprises:
determining a first set of preset mass-volume concentration values corresponding to trehalose included in the fibrinogen detection reagent;
determining a second set of preset mass volume concentration values corresponding to bovine serum albumin included in the fibrinogen detection reagent;
determining a third set of preset mass volume concentration values corresponding to arginine included in the fibrinogen detection reagent;
determining a fourth set of preset mass volume concentration values corresponding to urea included in the fibrinogen detection reagent;
determining a fifth set of preset volume concentration values corresponding to tissue factors included in the fibrinogen detection reagent;
determining a sixth set of preset mass volume concentration values corresponding to tirofiban included in the fibrinogen test reagent;
determining a seventh set of preset mass volume concentration values corresponding to sodium salicylate included in the fibrinogen detection reagent;
Determining an eighth set of preset mass volume concentration values corresponding to dipyridamole included in the fibrinogen detection reagent;
combining the fibrinogen detection reagents with different component concentration values according to the first preset mass volume concentration value set, the second preset mass volume concentration value set, the third preset mass volume concentration value set, the fourth preset mass volume concentration value set, the fifth preset volume concentration value set, the sixth preset mass volume concentration value set, the seventh preset mass volume concentration value set and the eighth preset mass volume concentration value set corresponding to the fibrinogen detection reagent;
according to the preset dosage, subpackaging and freeze-drying each fibrinogen detection reagent in the fibrinogen detection reagents to obtain fibrinogen detection reagent cups;
adding the preset dose of the calcium chloride solution to each of the respective fibrinogen detection reagent cups to obtain respective fibrinogen detection reagent cups containing the calcium chloride solution as respective target fibrinogen detection reagent cups;
Detecting the fibrinogen function of the target blood sample according to the target fibrinogen detection reagent cups to obtain detection results corresponding to the target fibrinogen detection reagent cups, wherein the target fibrinogen detection reagent cups in the target fibrinogen detection reagent cups correspond to detection results in the detection results;
and determining the concentration value of each component corresponding to the detection result meeting the preset result condition in each detection result as the concentration value of each component corresponding to each component in the fibrinogen detection reagent.
CN202311205488.7A 2023-09-15 2023-09-15 Preparation method of fibrinogen detection kit Pending CN117269507A (en)

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