CN112852917A - Titer determination equipment and method for defibrase - Google Patents

Titer determination equipment and method for defibrase Download PDF

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Publication number
CN112852917A
CN112852917A CN202011426645.3A CN202011426645A CN112852917A CN 112852917 A CN112852917 A CN 112852917A CN 202011426645 A CN202011426645 A CN 202011426645A CN 112852917 A CN112852917 A CN 112852917A
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frame body
defibrase
incubator
titer
motor
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杨延音
孙化富
胡雪琴
张亚红
潘婷立
王丽
胡世国
邵倩
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Chongqing Medical and Pharmaceutical College
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    • C12Q1/00Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
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    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61LMETHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
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    • B01PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
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    • B01D46/12Particle separators, e.g. dust precipitators, using filter plates, sheets or pads having plane surfaces in multiple arrangements
    • BPERFORMING OPERATIONS; TRANSPORTING
    • B01PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01DSEPARATION
    • B01D46/00Filters or filtering processes specially modified for separating dispersed particles from gases or vapours
    • B01D46/54Particle separators, e.g. dust precipitators, using ultra-fine filter sheets or diaphragms
    • B01D46/543Particle separators, e.g. dust precipitators, using ultra-fine filter sheets or diaphragms using membranes
    • BPERFORMING OPERATIONS; TRANSPORTING
    • B01PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01DSEPARATION
    • B01D53/00Separation of gases or vapours; Recovering vapours of volatile solvents from gases; Chemical or biological purification of waste gases, e.g. engine exhaust gases, smoke, fumes, flue gases, aerosols
    • B01D53/22Separation of gases or vapours; Recovering vapours of volatile solvents from gases; Chemical or biological purification of waste gases, e.g. engine exhaust gases, smoke, fumes, flue gases, aerosols by diffusion
    • B01D53/228Separation of gases or vapours; Recovering vapours of volatile solvents from gases; Chemical or biological purification of waste gases, e.g. engine exhaust gases, smoke, fumes, flue gases, aerosols by diffusion characterised by specific membranes
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61LMETHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L2209/00Aspects relating to disinfection, sterilisation or deodorisation of air
    • A61L2209/10Apparatus features
    • A61L2209/14Filtering means

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Abstract

The invention discloses titer measuring equipment for defibrase, which comprises an incubator, a culture dish, a purification mechanism, an air inlet pipe and an electric heating plate, wherein the incubator is used for holding a culture dish; a placing mechanism for placing a culture dish is arranged in the incubator, an electric heating plate is embedded on the inner wall of the incubator, a uniform heating assembly is arranged on the side wall of the incubator positioned on the electric heating plate, a purifying mechanism is arranged on the outer wall of the top surface of the incubator, the air outlet end of the purifying mechanism is connected with the air inlet end of an air inlet pipe, and the air outlet end of the air inlet pipe extends to the upper part of the inner cavity of the incubator; the first frame body, the second frame body, the third frame body, the fourth frame body, the fifth frame body and the sixth frame body are sequentially arranged side by side, and are provided with adaptive connecting holes, and connecting bolts are arranged in the connecting holes; the method has the advantages of improving the efficiency of defibrase culture and the accuracy of titer determination.

Description

Titer determination equipment and method for defibrase
Technical Field
The invention belongs to the technical field of defibrase, relates to titer measuring equipment and a titer measuring method, and particularly relates to defibrase titer measuring equipment and a titer measuring method.
Background
Defibrase is a proteolytic enzyme, and has effects in dissolving thrombus, inhibiting thrombosis, and improving microcirculation. The traditional Chinese medicine composition is mainly used for 1. acute cerebral infarction clinically; 2. improving various occlusive vascular diseases; 3. improving peripheral and microcirculation disturbance.
The comparison document CN211292573U discloses an absorbance monitoring device for an antibiotic titer tester, which relates to the technical field of antibiotic titer test and comprises a photometric machine body and a dissolving and debugging tank arranged above the photometric machine body, wherein a solution adding pipe is fixedly arranged in the middle of the upper end of the dissolving and debugging tank, a solvent adding pipe is fixedly arranged at one side of the upper end of the dissolving and debugging tank, which is close to the solution adding pipe, and liquid flow meters are fixedly arranged at the side positions of the solution adding pipe and the solvent adding pipe respectively, the liquid flow meters can monitor the adding amount of solution and solvent in real time, so as to accurately control the adding amount of solution and solvent, make the ratio of solution and solvent more accurate, and stir and mix the solution and the solvent mixed solution through a stirring rod and a stirring paddle, so that the solution and the solvent are mixed more quickly and uniformly, and improve the preparation quality of sample liquid, is favorable for improving the absorbance monitoring precision.
In the prior art, the titer of defibrase is measured by adopting an eye-measurement method, however, the titer of products is different due to different standards for determining a coagulation endpoint by testing human eyes, so that the titer of the products is influenced to a certain extent; in addition, the efficiency of culturing defibrase before measurement is low, which leads to a problem that the efficiency of defibrase measurement is low.
Disclosure of Invention
The invention aims to solve the problem that in the prior art, the titer of defibrase is measured by adopting a visual inspection method, but in the method, because the standards for judging the coagulation endpoint by eyes of a person are different, the titer of products is different, and the titer consistency of the products is influenced to a certain extent; in addition, the efficiency of culturing defibrase before measurement is low, which causes the problem that the efficiency of defibrase measurement is low, and a titer measuring apparatus and a titer measuring method for defibrase are provided.
The purpose of the invention can be realized by the following technical scheme:
a titer measuring device for defibrase comprises an incubator, a culture dish, a purification mechanism, an air inlet pipe and an electric heating plate;
a placing mechanism for placing a culture dish is arranged in the incubator, an electric heating plate is embedded on the inner wall of the incubator, a uniform heating assembly is arranged on the side wall of the incubator positioned on the electric heating plate, a purifying mechanism is arranged on the outer wall of the top surface of the incubator, the air outlet end of the purifying mechanism is connected with the air inlet end of an air inlet pipe, and the air outlet end of the air inlet pipe extends to the upper part of the inner cavity of the incubator;
the purification mechanism comprises a first frame body, a second frame body, an induced draft fan, a third frame body, a fourth frame body, a fifth frame body, a sixth frame body, a disinfectant tank, a first pipe body, a seventh frame body, a communicating pipe and a second pipe body; the first frame body, the second frame body, the third frame body, the fourth frame body, the fifth frame body and the sixth frame body are sequentially arranged side by side, and are provided with adaptive connecting holes, and connecting bolts are arranged in the connecting holes;
the first frame body is provided with an air inlet, two groups of draught fans are arranged in the second frame body in parallel, a first filter layer is arranged in the third frame body, a second filter layer is arranged in the fourth frame body, a plurality of groups of vent holes are uniformly formed in the fifth frame body, two groups of communicating pipes are arranged in the sixth frame body in parallel, the disinfectant box is located in the middle of the sixth frame body and the seventh frame body, disinfectant is loaded in the disinfectant box, a second pipe body is arranged on the side wall, close to the sixth frame body, of the disinfectant box, the air outlet end of the second pipe body extends to the position below the liquid level of the disinfectant, a first pipe body is arranged on the side wall, close to the seventh frame body, of the disinfectant box.
Preferably, the communicating pipe is composed of a first connecting pipe and a second connecting pipe, the first connecting pipe and the second connecting pipe are communicated with each other, the inner diameter of the first connecting pipe is larger than that of the second connecting pipe, and the second connecting pipe is sleeved with a sealing gasket and inserted into the second pipe body.
Preferably, the first filter layer is an activated carbon filter layer, the second filter layer is an air filter membrane, and the disinfectant is a sodium hypochlorite solution.
Preferably, the uniform heating component comprises a third motor, a shell, a connecting shaft, a driving bevel gear, a driven bevel gear, a rotating rod and fan blades; the third motor sets up on the top surface outer wall of incubator, the casing is installed on the lateral wall that the incubator is close to the electric heating board, the output and the connecting axle of third motor are connected, the connecting axle extends to the inside of casing, and rotate with the casing and be connected, the cover is equipped with multiunit initiative bevel gear on the connecting axle, initiative bevel gear is connected with driven bevel gear meshing, the one end and the driven bevel gear of bull stick are connected, the other end of bull stick passes the casing, and rotate with the casing and be connected, the other end and the flabellum of bull stick are connected.
Preferably, the placing mechanism comprises a base, a culture dish, a bottom plate, a first motor, a rotating shaft, an installation cylinder, a second motor, an installation plate and a placing assembly;
a bottom plate is arranged below the interior of the incubator, a first motor is mounted on the bottom plate, the output end of the first motor is connected with a rotating shaft, the rotating shaft penetrates through the bottom plate and is rotatably connected with the bottom plate, the top end of the rotating shaft is connected with an installation cylinder, and a plurality of groups of bases are uniformly arranged on the outer wall of the installation cylinder; the base is hollow structure, and installs the second motor in the base, and the output and the mounting panel of second motor are connected, and the mounting panel is located the top surface of base, and the mounting panel is connected with the culture dish through placing the subassembly, and the quantity of every row of base is the same with the quantity of flabellum, and the height of base is the same with the height that corresponds the flabellum.
Preferably, the bottom of the incubator is provided with a base, and the front of the incubator is provided with a chamber door.
Preferably, the placing assembly comprises a limiting groove, a limiting slide rod, a clamping block and a spring, the annular array is provided with a plurality of groups of limiting grooves on the mounting plate, the limiting slide rod is installed in the limiting groove, the clamping block is slidably arranged in the limiting groove, the limiting slide rod penetrates through the clamping block and is slidably connected with the clamping block, the spring is sleeved on the limiting slide rod, and two ends of the spring are respectively connected with the inner wall of the limiting groove and the clamping block.
A determination method of titer determination equipment of defibrase is characterized by comprising the following steps:
the first step is as follows: adding a culture medium comprising 5-6 parts by weight of peptone, 3-5 parts by weight of meat extract, 18-20 parts by weight of NaCl, 2-4 parts by weight of disodium hydrogen phosphate and 14-18 parts by weight of agar into a culture dish, starting an electric heating plate and a third motor to work, driving a connecting shaft to rotate by the third motor, driving fan blades to rotate by the connecting shaft under the meshing action of a driving bevel gear and a driven bevel gear, blowing heat generated by the electric heating plate to the culture medium, sterilizing the culture medium, cooling to room temperature after the treatment is finished, inoculating defibrase onto each group of culture dishes, starting a draught fan of a purifying mechanism to work, controlling the air to enter an air inlet pipe along a first frame body, a second frame body, a third frame body, a fourth frame body, a fifth frame body and a sixth frame body in sequence, and treating the air by a first filter layer, a second filter layer and a disinfectant to obtain sterile air, culturing defibrase under aseptic conditions, starting a second motor to work to drive a culture dish to rotate, and uniformly culturing the defibrase in an incubator;
the second step is that: opening the blood coagulation analyzer until the temperature reaches 37 ℃ for later use; dissolving defibrase with the titer of 100U/mL back, and diluting the defibrase with diluent to different concentrations to obtain a sample to be detected; respectively adding blood coagulation quality control plasma into sample tubes, placing in a detection part of a blood coagulation analyzer, and preheating at 37 deg.C for 2 min; measuring a sample to be measured and a standard solution, adding the sample to be measured and the standard solution into each sample tube, and respectively measuring under an analyzer; recording the solidification time; and judging the result, and calculating the titer of defibrase by adopting an external standard method.
Compared with the prior art, the invention has the beneficial effects that: adding a culture medium comprising 5-6 parts by weight of peptone, 3-5 parts by weight of meat extract, 18-20 parts by weight of NaCl, 2-4 parts by weight of disodium hydrogen phosphate and 14-18 parts by weight of agar into a culture dish 5, wherein an antibacterial ring obtained after the culture medium is cultured has clear edges, no double rings and no capsules, and has moderate diameter, so that the measurement error is greatly reduced, because a plurality of groups of bases are uniformly arranged on the outer wall of the mounting cylinder, a large amount of culture medium can be prepared at one time, then, an electric heating plate and a third motor are started to work, the third motor drives a connecting shaft to rotate, the connecting shaft drives fan blades to rotate through the meshing action of a driving bevel gear and a driven bevel gear, the heat generated by the electric heating plate is blown to the culture medium, the culture medium is sterilized, after the treatment is finished, the temperature is reduced to room temperature, and the defibrase is inoculated on each group of culture dishes, starting an induced draft fan of the purification mechanism to work, so that control enters an air inlet pipe along a first frame body, a second frame body, a third frame body, a fourth frame body, a fifth frame body and a sixth frame body in sequence, air is treated by a first filter layer, a second filter layer and disinfectant to obtain sterile air, defibrase is cultured under the sterile condition, and meanwhile, a second motor is started to work to drive a culture dish to rotate, so that the defibrase is uniformly cultured in a culture box; the culture box can be used for measuring the titer of defibrase, can provide a sterile environment for a culture medium, can store the effect of a plurality of culture media, does not damage the environment in the culture medium after gas is filtered and sterilized by the filter layer and the sterilization layer, provides a good environment for culture of the culture medium and the defibrase, and improves the titer efficiency of the defibrase;
opening the blood coagulation analyzer until the temperature reaches 37 ℃ for later use; dissolving defibrase with the titer of 100U/mL back, and diluting the defibrase with diluent to different concentrations to obtain a sample to be detected; respectively adding blood coagulation quality control plasma into sample tubes, placing in a detection part of a blood coagulation analyzer, and preheating at 37 deg.C for 2 min; measuring a sample to be measured and a standard solution, adding the sample to be measured and the standard solution into each sample tube, and respectively measuring under an analyzer; recording the solidification time; judging the result, and calculating the titer of defibrase by adopting an external standard method; the blood coagulation quality control plasma is used as a substrate, and the measurement is performed by a blood coagulation analyzer, so that the difference of titer among products caused by the difference of a tester on the determination standard of a coagulation endpoint is avoided, and the efficient and accurate measurement of the thrombin-like enzyme is realized.
Drawings
In order to facilitate understanding for those skilled in the art, the present invention will be further described with reference to the accompanying drawings.
FIG. 1 is a schematic structural diagram of the present invention.
FIG. 2 is a schematic view showing the inside structure of the incubator of the present invention.
Fig. 3 is an exploded perspective view of the purification mechanism of the present invention.
Fig. 4 is a schematic structural view showing a connection relationship between a sixth frame and a disinfectant tank according to the present invention.
FIG. 5 is a schematic view of a uniform heating element according to the present invention.
FIG. 6 is a schematic view showing the connection between the base and the culture dish according to the present invention.
Fig. 7 is a schematic structural view of a placement module according to the present invention.
In the figure: 1. a base; 2. an incubator; 3. a box door; 4. a base; 5. a culture dish; 6. a base plate; 7. a first motor; 8. uniformly heating the assembly; 9. a rotating shaft; 10. mounting the cylinder; 11. a limiting groove; 12. a limiting slide bar; 13. a second motor; 14. mounting a plate; 15. a clamping block; 16. a spring; 17. a purification mechanism; 18. an air inlet pipe; 19. a third motor; 20. a housing; 21. a connecting shaft; 22. a drive bevel gear; 23. a driven bevel gear; 24. a rotating rod; 25. a fan blade; 26. an electrical heating plate; 27. a first frame body; 28. a second frame body; 29. an induced draft fan; 30. a third frame body; 31. a fourth frame body; 32. a fifth frame body; 33. a sixth frame body; 34. a disinfectant tank; 35. a first pipe body; 36. a seventh frame body; 37. a communicating pipe; 38. a second tube.
Detailed Description
The technical solutions of the present invention will be described clearly and completely with reference to the following embodiments, and it should be understood that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
Example 1
Referring to fig. 1-7, a titer measuring apparatus for defibrase includes an incubator 2, a petri dish 5, a purification mechanism 17, an air inlet pipe 18, and an electric heating plate 26;
a placing mechanism for placing the culture dish 5 is arranged in the incubator 2, an electric heating plate 26 is embedded on the inner wall of the incubator 2, a uniform heating component 8 is arranged on the side wall of the incubator 2 positioned on the electric heating plate 26, a purifying mechanism 17 is arranged on the outer wall of the top surface of the incubator 2, the air outlet end of the purifying mechanism 17 is connected with the air inlet end of the air inlet pipe 18, and the air outlet end of the air inlet pipe 18 extends to the upper part of the inner cavity of the incubator 2;
the purification mechanism 17 includes a first frame 27, a second frame 28, an induced draft fan 29, a third frame 30, a fourth frame 31, a fifth frame 32, a sixth frame 33, a disinfectant tank 34, a first pipe 35, a seventh frame 36, a communication pipe 37, and a second pipe 38; the first frame body 27, the second frame body 28, the third frame body 30, the fourth frame body 31, the fifth frame body 32 and the sixth frame body 33 are sequentially arranged side by side, and the first frame body 27, the second frame body 28, the third frame body 30, the fourth frame body 31, the fifth frame body 32, the sixth frame body 33, the disinfectant tank 34 and the seventh frame body 36 are all provided with adaptive connecting holes, and connecting bolts are arranged in the connecting holes;
the first frame body 27 is provided with an air inlet, two groups of induced draft fans 29 are arranged in the second frame body 28 in parallel, a first filter layer is arranged in the third frame body 30, a second filter layer is arranged in the fourth frame body 31, a plurality of groups of vent holes are uniformly arranged in the fifth frame body 32, two groups of communicating pipes 37 are arranged in the sixth frame body 33 in parallel, the disinfectant tank 34 is positioned in the middle of the sixth frame body 33 and the seventh frame body 36, disinfectant is loaded in the disinfectant tank 34, a second pipe body 38 is arranged on the side wall of the disinfectant tank 34 close to the sixth frame body 33, the air outlet end of the second pipe body 38 extends to the position below the liquid level of the disinfectant, a first pipe body 35 is arranged on the side wall of the disinfectant tank 34 close to the seventh frame body 36, and the air inlet end.
The connection pipe 37 is composed of a first connection pipe and a second connection pipe, the first connection pipe and the second connection pipe are connected with each other, the inner diameter of the first connection pipe is larger than that of the second connection pipe, and the second connection pipe is sleeved with a sealing gasket and inserted into the second pipe body 38.
The first filter layer is an activated carbon filter layer, the second filter layer is an air filter membrane, and the disinfectant is a sodium hypochlorite solution.
The uniform heating component 8 comprises a third motor 19, a shell 20, a connecting shaft 21, a driving bevel gear 22, a driven bevel gear 23, a rotating rod 24 and fan blades 25; the third motor 19 sets up on the top surface outer wall of incubator 2, casing 20 is installed on the lateral wall that incubator 2 is close to electric heating board 26, the output and the connecting axle 21 of third motor 19 are connected, connecting axle 21 extends to the inside of casing 20, and rotate with casing 20 and be connected, the cover is equipped with multiunit drive bevel gear 22 on the connecting axle 21, drive bevel gear 22 is connected with driven bevel gear 23 meshing, the one end and the driven bevel gear 23 of bull stick 24 are connected, the other end of bull stick 24 passes casing 20, and rotate with casing 20 and be connected, the other end and the flabellum 25 of bull stick 24 are connected.
The placing mechanism comprises a base 4, a culture dish 5, a bottom plate 6, a first motor 7, a rotating shaft 9, an installation cylinder 10, a second motor 13, an installation plate 14 and a placing assembly;
a bottom plate 6 is arranged below the inner part of the incubator 2, a first motor 7 is arranged on the bottom plate 6, the output end of the first motor 7 is connected with a rotating shaft 9, the rotating shaft 9 penetrates through the bottom plate 6 and is rotatably connected with the bottom plate 6, the top end of the rotating shaft 9 is connected with an installation cylinder 10, and a plurality of groups of bases 4 are uniformly arranged on the outer wall of the installation cylinder 10; the base 4 is hollow structure, and installs the second motor 13 in the base 4, and the output and the mounting panel 14 of second motor 13 are connected, and the mounting panel 14 is located the top surface of base 4, and the mounting panel 14 is connected with culture dish 5 through placing the subassembly, and the quantity of every row of base 4 is the same with the quantity of flabellum 25, and the height of base 4 is the same with the height that corresponds flabellum 25.
The bottom of incubator 2 is provided with base 1, and the front of incubator 2 is provided with chamber door 3.
The placing assembly comprises a limiting groove 11, a limiting slide rod 12, clamping blocks 15 and springs 16, a plurality of groups of limiting grooves 11 are arranged on the mounting plate 14 in an annular array mode, the limiting slide rod 12 is installed in the limiting groove 11, the clamping blocks 15 are arranged in the limiting groove 11 in a sliding mode, the limiting slide rod 12 penetrates through the clamping blocks 15 and is connected with the clamping blocks 15 in a sliding mode, the springs 16 are sleeved on the limiting slide rod 12, and two ends of each spring 16 are connected with the inner wall of the limiting groove 11 and the clamping blocks 15 respectively.
A determination method of titer determination equipment of defibrase is characterized by comprising the following steps:
the first step is as follows: adding a culture medium comprising 5 parts by weight of peptone, 3 parts by weight of meat extract, 18 parts by weight of NaCl, 2 parts by weight of disodium hydrogen phosphate and 14 parts by weight of agar into a culture dish 5, then starting an electric heating plate 26 and a third motor 19 to work, driving a connecting shaft 21 to rotate by the third motor 19, driving fan blades 25 to rotate by the connecting shaft 21 under the meshing action of a driving bevel gear 22 and a driven bevel gear 23, blowing heat generated by the electric heating plate 26 to the culture medium, sterilizing the culture medium, cooling to room temperature after the treatment is finished, inoculating defibrase onto each group of culture dishes 5, starting a draught fan 29 of a purifying mechanism 17 to work, controlling the air to enter an air inlet pipe 18 along a first frame body 27, a second frame body 28, a third frame body 30, a fourth frame body 31, a fifth frame body 32 and a sixth frame body 33 in sequence, and obtaining sterile air after the air is treated by a first filter layer, a second filter layer and a sixth frame body 33, culturing defibrase under an aseptic condition, starting the second motor 13 to work, and driving the culture dish 5 to rotate so that the defibrase is uniformly cultured in the culture box 2;
the second step is that: opening the blood coagulation analyzer until the temperature reaches 37 ℃ for later use; dissolving defibrase with the titer of 100U/mL back, and diluting the defibrase with diluent to different concentrations to obtain a sample to be detected; respectively adding blood coagulation quality control plasma into sample tubes, placing in a detection part of a blood coagulation analyzer, and preheating at 37 deg.C for 2 min; measuring a sample to be measured and a standard solution, adding the sample to be measured and the standard solution into each sample tube, and respectively measuring under an analyzer; recording the solidification time; and judging the result, and calculating the titer of defibrase by adopting an external standard method.
Example 2
Compared with the embodiment 1, the difference is that the measuring method of the titer measuring equipment of defibrase is characterized by comprising the following steps:
the first step is as follows: adding a culture medium comprising 5.5 parts by weight of peptone, 4 parts by weight of meat extract, 19 parts by weight of NaCl, 3 parts by weight of disodium hydrogen phosphate and 16 parts by weight of agar into a culture dish 5, then starting an electric heating plate 26 and a third motor 19 to work, driving a connecting shaft 21 to rotate by the third motor 19, driving fan blades 25 to rotate by the connecting shaft 21 under the meshing action of a driving bevel gear 22 and a driven bevel gear 23, blowing heat generated by the electric heating plate 26 to the culture medium, sterilizing the culture medium, cooling to room temperature after the treatment is finished, inoculating defibrase onto each group of culture dishes 5, starting a draught fan 29 of a purifying mechanism 17 to work, controlling the air to enter a culture dish 18 along a first frame body 27, a second frame body 28, a third frame body 30, a fourth frame body 31, a fifth frame body 32 and a sixth frame body 33 in sequence, and treating the air by a first filter layer, a second filter layer and a disinfectant to obtain sterile air, culturing defibrase under an aseptic condition, starting the second motor 13 to work, and driving the culture dish 5 to rotate so that the defibrase is uniformly cultured in the culture box 2;
the second step is that: opening the blood coagulation analyzer until the temperature reaches 37 ℃ for later use; dissolving defibrase with the titer of 100U/mL back, and diluting the defibrase with diluent to different concentrations to obtain a sample to be detected; respectively adding blood coagulation quality control plasma into sample tubes, placing in a detection part of a blood coagulation analyzer, and preheating at 37 deg.C for 2 min; measuring a sample to be measured and a standard solution, adding the sample to be measured and the standard solution into each sample tube, and respectively measuring under an analyzer; recording the solidification time; and judging the result, and calculating the titer of defibrase by adopting an external standard method.
Example 3
Compared with the embodiment 1, the difference is that the measuring method of the titer measuring equipment of defibrase is characterized by comprising the following steps:
the first step is as follows: adding a culture medium comprising 6 parts by weight of peptone, 5 parts by weight of meat extract, 20 parts by weight of NaCl, 4 parts by weight of disodium hydrogen phosphate and 18 parts by weight of agar into a culture dish 5, then starting an electric heating plate 26 and a third motor 19 to work, driving a connecting shaft 21 to rotate by the third motor 19, driving fan blades 25 to rotate by the connecting shaft 21 under the meshing action of a driving bevel gear 22 and a driven bevel gear 23, blowing heat generated by the electric heating plate 26 to the culture medium, sterilizing the culture medium, cooling to room temperature after the treatment is finished, inoculating defibrase onto each group of culture dishes 5, starting a draught fan 29 of a purifying mechanism 17 to work, controlling the air to enter an air inlet pipe 18 along a first frame body 27, a second frame body 28, a third frame body 30, a fourth frame body 31, a fifth frame body 32 and a sixth frame body 33 in sequence, and obtaining sterile air after the air is treated by a first filter layer, a second filter layer and a sixth frame body 33, culturing defibrase under an aseptic condition, starting the second motor 13 to work, and driving the culture dish 5 to rotate so that the defibrase is uniformly cultured in the culture box 2;
the second step is that: opening the blood coagulation analyzer until the temperature reaches 37 ℃ for later use; dissolving defibrase with the titer of 100U/mL back, and diluting the defibrase with diluent to different concentrations to obtain a sample to be detected; respectively adding blood coagulation quality control plasma into sample tubes, placing in a detection part of a blood coagulation analyzer, and preheating at 37 deg.C for 2 min; measuring a sample to be measured and a standard solution, adding the sample to be measured and the standard solution into each sample tube, and respectively measuring under an analyzer; recording the solidification time; and judging the result, and calculating the titer of defibrase by adopting an external standard method.
The working principle of the invention is as follows: adding a culture medium comprising 5-6 parts by weight of peptone, 3-5 parts by weight of meat extract, 18-20 parts by weight of NaCl, 2-4 parts by weight of disodium hydrogen phosphate and 14-18 parts by weight of agar into a culture dish 5, wherein an antibacterial ring obtained after the culture of the culture medium has clear edges, no double rings and no capsules, and has moderate diameter, so that the measurement error is greatly reduced, because a plurality of groups of bases 4 are uniformly arranged on the outer wall of the mounting cylinder 10, a large amount of culture medium can be prepared at one time, then, an electric heating plate 26 and a third motor 19 are started to work, the third motor 19 drives a connecting shaft 21 to rotate, the connecting shaft 21 drives fan blades 25 to rotate through the meshing action of a driving bevel gear 22 and a driven bevel gear 23, the heat generated by the electric heating plate 26 is blown to the culture medium, the culture medium is sterilized, after the treatment, the temperature is reduced to room temperature, and defibrase is inoculated on each group of culture dishes 5, starting an induced draft fan 29 of the purification mechanism 17 to work, so that control enters the air inlet pipe 18 along the first frame body 27, the second frame body 28, the third frame body 30, the fourth frame body 31, the fifth frame body 32 and the sixth frame body 33 in sequence, the air is treated by the first filter layer, the second filter layer and the disinfectant to obtain sterile air, and when the defibrase is cultured under the sterile condition, the second motor 13 is started to work to drive the culture dish 5 to rotate, so that the defibrase is uniformly cultured in the culture box 2; the culture box 2 can be used for measuring the titer of defibrase, can provide a sterile environment for a culture medium, can store the effect of a plurality of culture media, does not damage the environment in the culture medium after gas is filtered and sterilized by the filter layer and the sterilization layer, provides a good environment for culture of the culture medium and the defibrase, and improves the titer of the defibrase;
opening the blood coagulation analyzer until the temperature reaches 37 ℃ for later use; dissolving defibrase with the titer of 100U/mL back, and diluting the defibrase with diluent to different concentrations to obtain a sample to be detected; respectively adding blood coagulation quality control plasma into sample tubes, placing in a detection part of a blood coagulation analyzer, and preheating at 37 deg.C for 2 min; measuring a sample to be measured and a standard solution, adding the sample to be measured and the standard solution into each sample tube, and respectively measuring under an analyzer; recording the solidification time; judging the result, and calculating the titer of defibrase by adopting an external standard method; the blood coagulation quality control plasma is used as a substrate, and the measurement is performed by a blood coagulation analyzer, so that the difference of titer among products caused by the difference of a tester on the determination standard of a coagulation endpoint is avoided, and the efficient and accurate measurement of the thrombin-like enzyme is realized.
The preferred embodiments of the invention disclosed above are intended to be illustrative only. The preferred embodiments are not intended to be exhaustive or to limit the invention to the precise forms disclosed. Obviously, many modifications and variations are possible in light of the above teaching. The embodiments were chosen and described in order to best explain the principles of the invention and the practical application, to thereby enable others skilled in the art to best utilize the invention. The invention is limited only by the claims and their full scope and equivalents.

Claims (8)

1. A titer of defibrase surveys equipment which characterized in that: comprises an incubator (2), a culture dish (5), a purification mechanism (17), an air inlet pipe (18) and an electric heating plate (26);
a placing mechanism for placing a culture dish (5) is arranged in the incubator (2), an electric heating plate (26) is embedded on the inner wall of the incubator (2), a uniform heating assembly (8) is arranged on the side wall of the incubator (2) positioned on the electric heating plate (26), a purifying mechanism (17) is arranged on the outer wall of the top surface of the incubator (2), the air outlet end of the purifying mechanism (17) is connected with the air inlet end of an air inlet pipe (18), and the air outlet end of the air inlet pipe (18) extends to the upper part of the inner cavity of the incubator (2);
the purification mechanism (17) comprises a first frame body (27), a second frame body (28), an induced draft fan (29), a third frame body (30), a fourth frame body (31), a fifth frame body (32), a sixth frame body (33), a disinfectant tank (34), a first pipe body (35), a seventh frame body (36), a communication pipe (37) and a second pipe body (38); the first frame body (27), the second frame body (28), the third frame body (30), the fourth frame body (31), the fifth frame body (32) and the sixth frame body (33) are sequentially arranged side by side, in addition, connecting holes which are matched with each other are formed in the first frame body (27), the second frame body (28), the third frame body (30), the fourth frame body (31), the fifth frame body (32), the sixth frame body (33), the disinfectant tank (34) and the seventh frame body (36), and connecting bolts are arranged in the connecting holes;
the first frame body (27) is provided with an air inlet, two groups of induced draft fans (29) are arranged in parallel in the second frame body (28), a first filter layer is arranged in the third frame body (30), a second filter layer is arranged in the fourth frame body (31), multiple groups of vent holes are uniformly arranged in the fifth frame body (32), two groups of communication pipes (37) are arranged in parallel in the sixth frame body (33), a disinfectant tank (34) is positioned in the middle of the sixth frame body (33) and the seventh frame body (36), disinfectant is loaded in the disinfectant tank (34), a second pipe body (38) is arranged on the side wall, close to the sixth frame body (33), of the disinfectant tank (34), the air outlet end of the second pipe body (38) extends to the position below the liquid level of the disinfectant, a first pipe body (35) is arranged on the side wall, close to the seventh frame body (36), and the air inlet end of the first pipe body (35) is positioned above the liquid level.
2. A titer measuring apparatus according to claim 1, wherein the communicating tube (37) is composed of a first connecting tube and a second connecting tube, the first connecting tube and the second connecting tube are communicated with each other, the inner diameter of the first connecting tube is larger than that of the second connecting tube, and the second connecting tube is sleeved with a sealing gasket and inserted into the second tube body (38).
3. The titer measuring apparatus for defibrase according to claim 1, wherein the first filter layer is an activated carbon filter layer, the second filter layer is an air filter membrane, and the disinfectant is a sodium hypochlorite solution.
4. A titer measuring apparatus for defibrase according to claim 1, wherein the uniform heating member (8) comprises a third motor (19), a housing (20), a connecting shaft (21), a driving bevel gear (22), a driven bevel gear (23), a rotating lever (24), and fan blades (25); third motor (19) set up on the top surface outer wall of incubator (2), casing (20) are installed on incubator (2) are close to the lateral wall of electric heating board (26), the output and connecting axle (21) of third motor (19) are connected, connecting axle (21) extend to the inside of casing (20), and rotate with casing (20) and be connected, the cover is equipped with multiunit initiative bevel gear (22) on connecting axle (21), initiative bevel gear (22) are connected with driven bevel gear (23) meshing, the one end and the driven bevel gear (23) of bull stick (24) are connected, casing (20) are passed to the other end of bull stick (24), and rotate with casing (20) and be connected, the other end and flabellum (25) of bull stick (24) are connected.
5. The titer reduction enzyme titer determination apparatus according to claim 1, wherein the placement mechanism comprises a base (4), a culture dish (5), a bottom plate (6), a first motor (7), a rotating shaft (9), a mounting cylinder (10), a second motor (13), a mounting plate (14), a placement component;
a bottom plate (6) is arranged below the inner part of the incubator (2), a first motor (7) is arranged on the bottom plate (6), the output end of the first motor (7) is connected with a rotating shaft (9), the rotating shaft (9) penetrates through the bottom plate (6) and is rotatably connected with the bottom plate (6), the top end of the rotating shaft (9) is connected with an installation cylinder (10), and a plurality of groups of bases (4) are uniformly arranged on the outer wall of the installation cylinder (10); base (4) are hollow structure, and install second motor (13) in base (4), and the output and mounting panel (14) of second motor (13) are connected, and mounting panel (14) are located the top surface of base (4), and mounting panel (14) are connected with culture dish (5) through placing the subassembly, and the quantity of every row of base (4) is the same with the quantity of flabellum (25), and the height of base (4) is the same with the height that corresponds flabellum (25).
6. A titer measuring apparatus for defibrase according to claim 1, wherein the bottom of the incubator (2) is provided with a base (1), and the front of the incubator (2) is provided with a door (3).
7. A titer of defibrase according to claim 5, wherein the placing component comprises a limiting groove (11), a limiting slide bar (12), a clamping block (15) and a spring (16), the mounting plate (14) is provided with a plurality of sets of limiting grooves (11) in an annular array, the limiting slide bar (12) is installed in the limiting groove (11), the clamping block (15) is slidably installed in the limiting groove (11), the limiting slide bar (12) passes through the clamping block (15) and is slidably connected with the clamping block (15), the spring (16) is sleeved on the limiting slide bar (12), and two ends of the spring (16) are respectively connected with the inner wall of the limiting groove (11) and the clamping block (15).
8. A measuring method of titer measuring apparatus of defibrase according to any one of claims 1 to 7, comprising the steps of:
the first step is as follows: adding a culture medium comprising, by weight, 5-6 parts of peptone, 3-5 parts of meat extract, 18-20 parts of NaCl, 2-4 parts of disodium hydrogen phosphate and 14-18 parts of agar into a culture dish (5), then starting an electric heating plate (26) and a third motor (19) to work, driving a connecting shaft (21) to rotate by the third motor (19), driving fan blades (25) to rotate by the connecting shaft (21) under the meshing action of a driving bevel gear (22) and a driven bevel gear (23), blowing heat generated by the electric heating plate (26) to the culture medium, sterilizing the culture medium, cooling to room temperature after the treatment is finished, inoculating defibrase onto each group of culture dishes (5), starting a draught fan (29) of a purification mechanism (17) to work, and controlling the culture medium to sequentially follow a first frame body (27), a second frame body (28), a third frame body (30), The fourth frame body (31), the fifth frame body (32) and the sixth frame body (33) enter the air inlet pipe (18), and the air is treated by the first filter layer, the second filter layer and the disinfectant to obtain sterile air, so that defibrase is cultured under the sterile condition, and meanwhile, the second motor (13) is started to work to drive the culture dish (5) to rotate, so that the defibrase is uniformly cultured in the culture box (2);
the second step is that: opening the blood coagulation analyzer until the temperature reaches 37 ℃ for later use; dissolving defibrase with the titer of 100U/mL back, and diluting the defibrase with diluent to different concentrations to obtain a sample to be detected; respectively adding blood coagulation quality control plasma into sample tubes, placing in a detection part of a blood coagulation analyzer, and preheating at 37 deg.C for 2 min; measuring a sample to be measured and a standard solution, adding the sample to be measured and the standard solution into each sample tube, and respectively measuring under an analyzer; recording the solidification time; and judging the result, and calculating the titer of defibrase by adopting an external standard method.
CN202011426645.3A 2020-12-09 2020-12-09 Titer determination equipment and method for defibrase Pending CN112852917A (en)

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Application publication date: 20210528