Rapid diagnosis instrument for rabies virus and method thereof

Abstract

The invention discloses a rapid diagnosis instrument for rabies virus and a method thereof, and relates to the technical field of rabies virus determination.The rapid diagnosis instrument comprises a shell, one end of the shell is fixedly connected with a movable handle, the edge of the other end of the shell is rotationally connected with a rotating cover, and the bottom of the shell is fixedly connected with two supporting strips; the bottom of the inner wall of the shell is fixedly connected with a base, the interior of the base is rotationally connected with a rotating cylinder, one end of the rotating cylinder is fixedly connected with a rotating disc, and the upper surface of the shell is rotationally connected with a first sealing cover, a second sealing cover and a third sealing cover. According to the rapid diagnosis instrument for rabies virus disclosed by the invention, light-emitting cells in a plurality of culture holes in the upper surface of a 96-well plate are photographed through a camera, the number of cells in mixed solutions with different concentrations is counted through an image recognition technology, and the half infection dose of BHK cells is calculated through a Reed-Muench method; the effect of rapidly determining the infection amount of the rabies virus is achieved.

Description

Technical Field

[0001] The present invention relates to the technical field of rabies virus determination, in particular to a rapid diagnostic instrument and method for rabies virus. Background Art

[0002] Rabies virus monitoring is crucial to public health, and the technology for detecting viral content has undergone significant development. The mouse method, as a traditional detection method, is based on the principle of inoculating rabies virus into mice and evaluating the pathogenicity and content of the virus by observing the incidence and mortality of mice. The application of this method provides an important basis for the epidemic monitoring of rabies and the evaluation of vaccine efficacy. However, with the increasing demand for detection accuracy and efficiency, alternative detection technologies such as molecular biology methods and cell culture methods are also gradually gaining attention.

[0003] Due to the high cost of the mouse method, including the cost of purchasing mice, raising and managing them, factors such as the mouse's physique, health status and breeding environment will affect the test results, leading to deviations in the results. This biological detection method has a large variability and may produce inconsistent results, making the determination of viral content unstable. The mouse method has a long test cycle and usually takes several weeks to obtain results, which is particularly inconvenient in emergency situations. Raising mice also brings about high energy and resource consumption, which cannot meet actual needs. Summary of the Invention

[0004] The invention discloses a rapid diagnostic instrument and method for rabies virus, aiming to solve the technical problems raised in the technical background.

[0005] In order to achieve the above object, the present invention adopts the following technical solutions: A rapid diagnostic instrument for rabies virus, comprising a shell, one end of the shell being fixedly connected to a mobile handle, an edge of the other end of the shell being rotatably connected to a rotating cover, the bottom of the shell being fixedly connected to two support bars, the bottom of the inner wall of the shell being fixedly connected to a base, the interior of the base being rotatably connected to a rotating cylinder, one end of the rotating cylinder being fixedly connected to a rotating disk, a first sealing cover, a second sealing cover, and a third sealing cover being rotatably connected to the upper surface of the shell, a ventilator being fixedly connected to the top of the inner wall of the shell, a plurality of filter plates being provided inside the ventilator, a plurality of air inlets being provided on the upper surface of the ventilator, a plurality of air inlet slits being provided on the inner side surface of the ventilator, a controller being provided on the upper surface of the controller, a button and a USB interface being provided on the upper surface of the controller, and a camera and an ultraviolet lamp being provided at the bottom of the third sealing cover; A collecting mechanism is installed at the bottom of the rotating cover, and the collecting mechanism is used to collect various experimental tools used for detecting rabies virus; A constant temperature mechanism is installed inside the housing, and the constant temperature mechanism is used to control the temperature inside the housing and replace the air inside the housing; An automatic maintaining mechanism is installed inside the housing near the constant temperature mechanism, and the automatic maintaining mechanism cooperates with the constant temperature mechanism to culture BHK cells; A plurality of locking mechanisms are installed on the upper surface of the shell near the rotating drum, and the locking mechanisms are used to lock the first sealing cover, the second sealing cover and the third sealing cover to maintain the sealing of the interior of the shell.

[0006] The collecting mechanism includes a maintenance liquid tank arranged inside the shell at the bottom of the rotating cover, and the interior of the shell near the maintenance liquid tank is respectively provided with a cell cylinder, a virus cylinder, a maintenance liquid cylinder, a fluorescent liquid cylinder and a test tube warehouse, and the interior of the shell near the test tube warehouse is provided with two pipettes on one side, the lower surfaces of the first sealing cover, the second sealing cover and the third sealing cover are respectively provided with temperature sensors, the upper surfaces of the first sealing cover, the second sealing cover and the third sealing cover are respectively provided with pulling blocks, and the connections between the first sealing cover, the second sealing cover and the third sealing cover and the shell are respectively provided with torsion springs.

[0007] The constant temperature mechanism includes an air pump fixedly connected to the bottom of the inner wall of the shell, an auxiliary pipe and an air outlet pipe are respectively provided on one side of the air pump, a three-way valve is provided at one end of the air outlet pipe, a heating pipe is provided on one side of the three-way valve, the top of the heating box is fixedly connected to a mounting shell, the heating pipe is located inside the mounting shell, and an ultraviolet lamp is provided at one end of the heating pipe.

[0008] A connecting pipe is provided at the bottom of the three-way valve, a heating box is provided at the bottom of the inner wall of the outer shell, a plurality of heating plates are provided inside the heating box, a cleaning cylinder is provided between the plurality of heating plates, a plurality of deceleration plates are provided inside the cleaning cylinder, and one end of the connecting pipe is connected to one end of the cleaning cylinder.

[0009] The other end of the cleaning cylinder is provided with a discharge pipe, one end of the ultraviolet lamp passes through the ventilation cylinder, a plurality of ventilation holes are provided on the outer wall of the rotating cylinder, a hexagonal groove is provided on the top of the rotating cylinder, and a rotating handle is provided on the top of the hexagonal groove.

[0010] In a preferred embodiment, the automatic maintaining mechanism includes a micro pump fixedly connected to the bottom of the inner wall of the shell, a suction pipe and a discharge pipe are respectively provided on one side of the micro pump, a connecting pipe is provided at the bottom of the first sealing cover, and a plurality of dripping tubes are provided at the bottom of the connecting tube, and a solenoid valve is respectively provided at one end of the dripping tube.

[0011] A ninety-six-hole plate is provided on the upper surface of the rotating disk at the bottom of the connecting tube. A plurality of culture holes are provided on the upper surface of the ninety-six-hole plate. The number of culture holes and dropper tubes is the same. One end of the drainage tube is connected to the interior of the connecting tube.

[0012] In a preferred solution, the locking mechanism includes a plurality of blocks slidably connected to the upper surface of the housing near the rotating cylinder, a first spring is provided on one side of the block, and a middle groove is provided inside the block.

[0013] The upper surface of the shell is slidably connected to a mounting rod inside the middle groove, the top of the mounting rod is provided with a pressing button, the bottom of the mounting rod is provided with a second spring, and the bottom of the pressing button is provided with a limiting block.

[0014] A method for using a rapid diagnostic instrument for rabies virus comprises the following steps: S1. Preparation: Rotate the first sealing cover 90 degrees. First, place the 96-well plate on the rotating disk at the bottom of the first sealing cover. The upper surface of the rotating disk is provided with a plurality of stoppers for fixing the 96-well plate. The stoppers are used to fix the position of the 96-well plate. Use a pipette to drop the cell solution inside the cell cylinder into the multiple culture wells on the upper surface of the 96-well plate. S2. Culture: Take out multiple test tubes from the test tube compartment and use a pipette to dilute the rabies virus stock solution in the virus cartridge and the cell culture maintenance solution in the maintenance solution cartridge proportionally into ten levels of concentration. Since there are culture wells on the upper surface of a 96-well plate, drop each concentration of the mixed solution into eight culture wells vertically. Drop ten concentrations of the mixed solution into each of the twelve columns from left to right. Drop only the maintenance solution into the eleventh and twelfth columns as blank controls. Culture for four to five days. S3. Staining: Rotate the rotating cylinder and rotating disk by turning the handle to rotate the 96-well plate from the bottom of the first sealing cover to the bottom of the second sealing cover. Rotate the second sealing cover to expose the upper surface of the 96-well plate. Use a pipette to drop the fluorescent liquid in the fluorescent liquid cylinder into the culture wells to perform fluorescent staining on the BHK cells. Let it stand for one hour. S4. Counting: A 96-well plate is rotated from the bottom of the second sealing cover to the bottom of the third sealing cover. The ultraviolet light emitted by the ultraviolet lamp causes the interior of the BHK cells infected with rabies virus to fluoresce. A camera is used to photograph the luminescent cells in multiple culture wells on the upper surface of the 96-well plate. The number of cells in the mixed solution of different concentrations is counted using image recognition technology, and the median infection dose of the BHK cells is calculated using the Reed-Muench method.

[0015] From the above, it can be seen that the rapid diagnostic instrument for rabies virus provided by the present invention has the following technical effects.

[0016] First, ultraviolet light is emitted from an ultraviolet lamp to cause the interior of the BHK cells infected with rabies virus to fluoresce. A camera is used to photograph the luminescent cells inside multiple culture wells on the surface of a 96-well plate. Image recognition technology is used to count the number of cells in the mixed solution of different concentrations, and the half-infectious dose of BHK cells is calculated using the Reed-Muench method. Compared with the more than fourteen days required to calculate the half-infectious dose of rabies virus in BHK cells using experiments on mice, it only takes four to five days to determine the half-infectious dose of rabies virus in BHK cells, which has the effect of quickly determining the half-infectious dose of rabies virus.

[0017] Secondly, since the virus invades cells slowly, it takes four to five days to culture. During the culture process, maintenance fluid needs to be continuously dripped into the culture wells to ensure that the cells remain alive. At regular intervals, the controller controls the micropump to extract maintenance fluid from the inside of the maintenance fluid tank, and opens the solenoid valve on the drip tube to drip the maintenance fluid into multiple culture wells on the upper surface of the 96-well plate, thereby realizing automatic culture of BHK cells and reducing manual participation in the culture of BHK cells.

[0018] Third: The flow of air entering the heating tube is controlled by a three-way valve, so that part of the air enters the interior of the heating tube. The air inside the heating tube is heated by the heating plate. The heated air is discharged into the interior of the shell through the ultraviolet lamp, thereby increasing the temperature inside the shell. The temperature inside the shell is detected by a temperature sensor, so that the ambient temperature inside the shell is maintained at 37 degrees, thereby utilizing the residual heat from the heating plate to kill the rabies virus and maintain the optimal culture temperature of BHK cells. BRIEF DESCRIPTION OF THE DRAWINGS

[0019] Figure 1 This is a schematic diagram of the top view of the structure proposed by the present invention.

[0020] Figure 2 This is a bottom-up structural schematic diagram of the present invention.

[0021] Figure 3 This is a structural schematic diagram of the rotating cover in the rotating state proposed by the present invention.

[0022] Figure 4 This is a schematic cross-sectional structural diagram of the present invention.

[0023] Figure 5 This is a schematic structural diagram of the first sealing cover in the rotating state proposed by the present invention.

[0024] Figure 6 This is a schematic diagram of the internal structure proposed by the present invention.

[0025] Figure 7 This is a schematic diagram of the local structure proposed by the present invention.

[0026] Figure 8 The present invention proposes Figure 7 Schematic diagram of the enlarged structure of point A.

[0027] In the figure: 1. housing; 2. rotating cover; 3. first sealing cover; 4. second sealing cover; 5. controller; 6. third sealing cover; 7. moving handle; 8. support bar; 9. discharge pipe; 10. pulling block; 11. cell cylinder; 12. maintenance liquid tank; 13. virus cylinder; 14. maintenance liquid cylinder; 15. fluorescent liquid cylinder; 16. test tube compartment; 17. pipette; 18. connecting pipe; 19. 96-well plate; 20. air inlet slit; 21. vent cylinder; 22. heating box; 23. air pump; 24 , rotating cylinder; 25. base; 26. micro pump; 27. suction tube; 28. discharge tube; 29. ​​rotating handle; 30. temperature sensor; 31. camera; 32. ultraviolet lamp; 33. heating tube; 34. heating plate; 35. cleaning cylinder; 36. deceleration plate; 37. mounting shell; 38. three-way valve; 39. auxiliary pipe; 40. exhaust pipe; 41. press button; 42. first spring; 43. block; 44. second spring; 45. mounting rod; 46. limiting block; 47. air inlet. DETAILED DESCRIPTION

[0028] The technical solutions in the embodiments of the present invention will be clearly and completely described below in conjunction with the drawings in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, rather than all the embodiments.

[0029] Reference Figure 1 — Figure 8 , a rapid diagnostic instrument for rabies virus, comprising a shell 1, one end of the shell 1 is fixedly connected to a mobile handle 7, the edge of the other end of the shell 1 is rotatably connected to a rotating cover 2, the bottom of the shell 1 is fixedly connected to two support bars 8, the bottom of the inner wall of the shell 1 is fixedly connected to a base 25, the inside of the base 25 is rotatably connected to a rotating cylinder 24, one end of the rotating cylinder 24 is fixedly connected to a rotating disk, the upper surface of the shell 1 is rotatably connected to a first sealing cover 3, a second sealing cover 4 and a third sealing cover 6, the top of the inner wall of the shell 1 is fixedly connected to a ventilator 21, the interior of the ventilator 21 is provided with a plurality of filter plates, the upper surface of the ventilator 21 is provided with a plurality of air inlets 47, the inner side surface of the ventilator 21 is provided with a plurality of air inlet slits 20, the upper surface of the third sealing cover 6 is provided with a controller 5, the upper surface of the controller 5 is provided with a button and a USB interface, and the bottom of the third sealing cover 6 is provided with a camera 31 and an ultraviolet lamp 32; A collecting mechanism is installed at the bottom of the rotating cover 2, which is used to collect various experimental tools used for detecting rabies virus; A constant temperature mechanism is installed inside the housing 1, which is used to control the temperature inside the housing 1 and replace the air inside the housing 1; An automatic maintaining mechanism is installed inside the housing 1 near the constant temperature mechanism, and the automatic maintaining mechanism cooperates with the constant temperature mechanism to culture BHK cells; A plurality of locking mechanisms are installed on the upper surface of the housing 1 near the rotating cylinder 24 , and the locking mechanisms are used to lock the first sealing cover 3 , the second sealing cover 4 and the third sealing cover 6 to maintain the sealing inside the housing 1 .

[0030] The collecting mechanism includes a maintenance liquid tank 12 arranged inside the shell 1 at the bottom of the rotating cover 2, and a cell cylinder 11, a virus cylinder 13, a maintenance liquid cylinder 14, a fluorescent liquid cylinder 15 and a test tube warehouse 16 are respectively provided in the interior of the shell 1 near the maintenance liquid tank 12. Two pipettes 17 are provided on one side of the interior of the shell 1 near the test tube warehouse 16, and the lower surfaces of the first sealing cover 3, the second sealing cover 4 and the third sealing cover 6 are respectively provided with temperature sensors 30, and the upper surfaces of the first sealing cover 3, the second sealing cover 4 and the third sealing cover 6 are respectively provided with pulling blocks 10, and the connections between the first sealing cover 3, the second sealing cover 4 and the third sealing cover 6 and the shell 1 are respectively provided with torsion springs.

[0031] The constant temperature mechanism includes an air pump 23 fixedly connected to the bottom of the inner wall of the outer shell 1, and an auxiliary pipe 39 and an air outlet pipe 40 are respectively provided on one side of the air pump 23. A three-way valve 38 is provided at one end of the air outlet pipe 40, and a heating pipe 33 is provided on one side of the three-way valve 38. The top of the heating box 22 is fixedly connected to a mounting shell 37, and the heating pipe 33 is located inside the mounting shell 37. An ultraviolet lamp 32 is provided at one end of the heating pipe 33.

[0032] A connecting pipe is provided at the bottom of the three-way valve 38, a heating box 22 is provided at the bottom of the inner wall of the outer shell 1, a plurality of heating plates 34 are provided inside the heating box 22, a cleaning cylinder 35 is provided between the plurality of heating plates 34, a plurality of deceleration plates 36 are provided inside the cleaning cylinder 35, and one end of the connecting pipe is connected to one end of the cleaning cylinder 35.

[0033] The other end of the cleaning cylinder 35 is provided with a discharge pipe 9, one end of the ultraviolet lamp 32 passes through the ventilation cylinder 21, a plurality of ventilation holes are provided on the outer wall of the rotating cylinder 24, a hexagonal groove is provided on the top of the rotating cylinder 24, and a rotating handle 29 is provided on the top of the hexagonal groove.

[0034] It is further explained that the experimental tools include a cell cartridge 11 , a virus cartridge 13 , a maintenance liquid cartridge 14 , a fluorescent liquid cartridge 15 , a test tube compartment 16 , a test tube, and a pipette 17 .

[0035] In the embodiment, the first sealing cover 3 is rotated by ninety degrees, and the ninety-six hole plate 19 is first placed on the rotating disc at the position of the bottom of the first sealing cover 3. The upper surface of the rotating disc is provided with a plurality of limiting blocks for fixing the ninety-six hole plate 19. The limiting blocks are used to fix the position of the ninety-six hole plate 19. The pipette 17 is used to drop the cell liquid in the cell cylinder 11 into a plurality of culture holes on the upper surface of the ninety-six hole plate 19.

[0036] Further, a plurality of test tubes are taken out from the inside of the test tube bin 16. The rabies virus stock solution in the virus cylinder 13 and the cell culture maintenance liquid in the maintenance liquid cylinder 14 are diluted in proportion by using the pipette 17. The dilution is ten levels of concentration. Since there are 96 culture holes on the upper surface of the ninety-six hole plate 19, and the 96 culture holes are arranged in the form of eight rows and twelve columns, the mixed liquid of each concentration is respectively dropped into eight culture holes in the longitudinal direction. Twelve columns are used to respectively drop the mixed liquid of ten concentrations from left to right. The eleventh column and the twelfth column are respectively used to drop only the maintenance liquid as a blank control group.

[0037] In the embodiment, in the process of culturing cells, the air pump 23 is controlled by the controller 5 to form negative pressure in the inside of the base 25 by using the auxiliary pipe 39. The air is sucked into the inside of the ventilation cylinder 21 through the air inlet 47. The impurities and dust in the air are removed by filtering through a plurality of filter plates in the inside of the ventilation cylinder 21. The air enters the inside of the shell 1 through the air inlet slit 20. The air flows above the ninety-six hole plate 19 to provide sufficient oxygen for cell culture. The air enters the inside of the base 25 through the ventilation holes on the rotating cylinder 24. The air enters the inside of the cleaning cylinder 35 through the air pump 23 and the air outlet pipe 40.

[0038] Further, the plurality of heating plates 34 are controlled by the controller 5 to heat the cleaning cylinder 35. The temperature in the inside of the base 25 reaches more than two hundred and ten degrees. When the air passes through the plurality of speed reduction plates 36 in the inside of the cleaning cylinder 35, the speed reduction plates 36 reduce the flow speed of the air. The temperature in the inside of the base 25 kills the rabies virus mixed in the air. Finally, the air is discharged from the inside of the shell 1 through the discharge pipe 9. The effect of exchanging gas for cell culture and removing the rabies virus in the air is achieved. Further, the flow of air entering the heating pipe 33 is controlled by the three-way valve 38. Part of the air enters the inside of the heating pipe 33. The air in the inside of the heating pipe 33 is heated by the heating plate 34. The heated air is discharged into the inside of the shell 1 through the ultraviolet lamp 32. The temperature in the inside of the shell 1 is increased. The temperature in the inside of the shell 1 is detected by the temperature sensor 30. The ambient temperature in the inside of the shell 1 is maintained at thirty-seven degrees. The effect of maintaining the optimal culture temperature of BHK cells by the residual heat of the heating plate 34 to kill the rabies virus is achieved.

[0039] Reference Figure 1 , Figure 4 ,Figure 5 and Figure 7 In a preferred embodiment, the automatic maintaining mechanism includes a micro pump 26 fixedly connected to the bottom of the inner wall of the shell 1, and a liquid suction pipe 27 and a liquid discharge pipe 28 are respectively provided on one side of the micro pump 26. A connecting pipe 18 is provided at the bottom of the first sealing cover 3, and a plurality of dripping tubes are provided at the bottom of the connecting pipe 18, and a solenoid valve is provided at one end of each dripping tube.

[0040] A ninety-six-hole plate 19 is provided on the upper surface of the rotating disk at the bottom of the connecting tube 18. A plurality of culture holes are provided on the upper surface of the ninety-six-hole plate 19. The number of culture holes and dropper tubes is the same. One end of the drainage tube 28 is connected to the interior of the connecting tube 18.

[0041] In this embodiment, since the virus invades cells slowly, it takes four to five days to culture. During the culture process, maintenance fluid needs to be continuously dripped into the interior of the culture wells to ensure that the cells remain alive. At regular intervals, the controller 5 controls the micropump 26 to extract maintenance fluid from the interior of the maintenance fluid tank 12, and by opening the solenoid valve on the drip tube, the maintenance fluid is dripped into the interior of multiple culture wells on the upper surface of the 96-well plate 19, thereby realizing automatic culture of BHK cells and reducing manual participation in the culture of BHK cells.

[0042] What needs to be explained further is that by rotating the handle 29 to rotate the rotating cylinder 24 and the rotating disk, the ninety-six-well plate 19 is rotated from the bottom of the first sealing cover 3 to the bottom of the second sealing cover 4, and the second sealing cover 4 is rotated to expose the upper surface of the ninety-six-well plate 19. The fluorescent liquid inside the fluorescent liquid cylinder 15 is dripped into the inside of the culture well using a pipette 17 to perform fluorescent staining on the BHK cells.

[0043] Furthermore, after standing for one hour, the rotating cylinder 24 and the rotating disk are rotated by turning the handle 29 to rotate the ninety-six-well plate 19 from the bottom of the second sealing cover 4 to the bottom of the third sealing cover 6. The ultraviolet lamp 32 is turned on by the controller 5. The ultraviolet lamp 32 emits ultraviolet rays to make the part of the BHK cells infected with rabies virus inside emit fluorescence. The camera 31 is used to take pictures of the luminous cells inside multiple culture wells on the upper surface of the ninety-six-well plate 19. The number of cells in the mixed solution of different concentrations is counted by image recognition technology, and the half-infectious dose of BHK cells is calculated by the Reed-Muench method. Compared with the half-infectious dose of rabies virus of BHK cells calculated by the mouse experiment, which takes more than fourteen days, it only takes four to five days to determine the half-infectious dose of rabies virus using BHK cells, which has the effect of quickly determining the half-infectious dose of rabies virus.

[0044] Reference Figure 1 、 Figure 2 、 Figure 3 and Figure 8In a preferred embodiment, the locking mechanism includes a plurality of blocks 43 slidably connected to the upper surface of the housing 1 near the rotating cylinder 24, a first spring 42 is provided on one side of the block 43, and an intermediate groove is provided inside the block 43.

[0045] A mounting rod 45 is slidably connected to the interior of the middle groove on the upper surface of the housing 1 , a pressing button 41 is provided on the top of the mounting rod 45 , a second spring 44 is provided at the bottom of the mounting rod 45 , and a limiting block 46 is provided at the bottom of the pressing button 41 .

[0046] In this embodiment, by rotating the first sealing cover 3, the second sealing cover 4 and the third sealing cover 6, the edges of the first sealing cover 3, the second sealing cover 4 and the third sealing cover 6 squeeze the clamping block 43, and the clamping block 43 squeezes the first spring 42, so that the clamping block 43 is clamped above the edges of the first sealing cover 3, the second sealing cover 4 and the third sealing cover 6, thereby fixing the positions of the first sealing cover 3, the second sealing cover 4 and the third sealing cover 6. When it is necessary to rotate the first sealing cover 3, the second sealing cover 4 and the third sealing cover 6, the press button 41 is pressed, and the clamping block 43 is squeezed by the limiting block 46, so that the clamping block 43 slides to one side, thereby allowing the first sealing cover 3, the second sealing cover 4 and the third sealing cover 6 to rotate.

[0047] Working principle: When in use, rotate the first sealing cover 3 ninety degrees, first place the ninety-six-well plate 19 on the rotating disk at the bottom of the first sealing cover 3, and the upper surface of the rotating disk is provided with a plurality of limit blocks for fixing the ninety-six-well plate 19. The limit blocks are used to fix the position of the ninety-six-well plate 19, and the cell liquid inside the cell cylinder 11 is dripped into the multiple culture wells on the upper surface of the ninety-six-well plate 19 using a pipette 17. Take out multiple test tubes from the inside of the test tube warehouse 16, and use the pipette 17 to dilute the rabies virus stock solution inside the virus cylinder 13 and the cell culture maintenance solution inside the maintenance liquid cylinder 14 in proportion to ten levels of concentration. Since there are 96 culture wells on the upper surface of the ninety-six-well plate 19, and the 96 culture wells are arranged in eight rows and twelve columns, The mixture of each concentration is respectively dripped into eight culture wells in the vertical direction. The mixture of ten concentrations is dripped into the twelve columns from left to right. The eleventh column and the twelfth column are only dripped with the maintenance solution as a blank control group. Since the virus invades the cells slowly, it needs to be cultured for four to five days. During the culture process, it is necessary to continuously drip the maintenance solution into the inside of the culture well to ensure that the cells remain alive. At regular intervals, the controller 5 controls the micro pump 26 to extract the maintenance solution from the inside of the maintenance solution tank 12, and opens the solenoid valve on the dropper to drip the maintenance solution into the multiple culture wells on the upper surface of the ninety-six-well plate 19, thereby realizing automatic culture of BHK cells and reducing the effect of manual participation in the culture of BHK cells. In the process of culturing cells, by controlling The controller 5 controls the air pump 23 to use the auxiliary pipe 39 to form a negative pressure inside the base 25, and sucks air into the interior of the ventilation cylinder 21 through the air inlet 47. The air is filtered by multiple filter plates inside the ventilation cylinder 21 to remove impurities and dust in the air. The air enters the interior of the shell 1 through the air inlet slit 20, and the air flows over the top of the ninety-six-well plate 19, providing sufficient oxygen for cell culture. The air enters the interior of the base 25 through the vent holes on the rotating cylinder 24, and enters the interior of the cleaning cylinder 35 through the air pump 23 and the outlet pipe 40. The controller 5 controls the multiple heating plates 34 to heat the cleaning cylinder 35, so that the internal temperature of the base 25 reaches above 210 degrees. When the air passes through the multiple deceleration plates 36 inside the cleaning cylinder 35, the deceleration plates 36 reduce the air pressure. The flow rate of the air and the temperature inside the base 25 kill the rabies virus mixed in the air. Finally, the air is discharged from the interior of the housing 1 through the exhaust pipe 9, which has the effect of exchanging gas for cell culture and clearing the rabies virus in the air. The flow rate of air entering the heating pipe 33 is controlled by the three-way valve 38, so that part of the air enters the interior of the heating pipe 33. The air inside the heating pipe 33 is heated by the heating plate 34. The heated air is discharged into the interior of the housing 1 through the ultraviolet lamp 32, thereby increasing the temperature inside the housing 1. The temperature inside the housing 1 is detected by the temperature sensor 30, so that the ambient temperature inside the housing 1 is maintained at 37 degrees. The residual heat from the heating plate 34 that kills the rabies virus is used to maintain the optimal culture temperature of BHK cells.The handle 29 is used to rotate the rotating cylinder 24 and the rotating disk so that the 96-well plate 19 is rotated from the bottom of the first sealing cover 3 to the bottom of the second sealing cover 4. The second sealing cover 4 is rotated to expose the upper surface of the 96-well plate 19. The fluorescent liquid in the fluorescent liquid cylinder 15 is dripped into the interior of the culture well using a pipette 17 to perform fluorescent staining on the BHK cells. After standing for one hour, the handle 29 is used to rotate the rotating cylinder 24 and the rotating disk so that the 96-well plate 19 is rotated from the bottom of the second sealing cover 4 to the bottom of the third sealing cover 6. The controller 5 controls the ultraviolet lamp 32 to turn on. The ultraviolet lamp 32 emits ultraviolet light to cause the rabies virus-infected parts of the BHK cells to emit fluorescence. The camera 31 takes pictures of the luminous cells in multiple culture wells on the upper surface of the 96-well plate 19. The number of cells in the mixed solution of different concentrations is counted using image recognition technology, and the number of BHK cells is calculated using the Reed-Muench method. Compared with the 50% infection dose of rabies virus calculated by the BHK cell experiment using mice, which takes more than fourteen days, the 50% infection dose of rabies virus determined using BHK cells only takes four to five days, which has the effect of quickly determining the 50% infection dose of rabies virus. By rotating the first sealing cover 3, the second sealing cover 4, and the third sealing cover 6, the edges of the first sealing cover 3, the second sealing cover 4, and the third sealing cover 6 squeeze the card block 43, and the card block 43 squeezes the first spring 42, so that the card block 43 is stuck above the edge of the first sealing cover 3, the second sealing cover 4, and the third sealing cover 6, which has the effect of fixing the position of the first sealing cover 3, the second sealing cover 4, and the third sealing cover 6. When it is necessary to rotate the first sealing cover 3, the second sealing cover 4, and the third sealing cover 6, the button 41 is pressed, and the card block 43 is squeezed by the limiting block 46, so that the card block 43 slides to one side, thereby allowing the first sealing cover 3, the second sealing cover 4, and the third sealing cover 6 to rotate.

[0048] The above description is only a preferred specific embodiment of the present invention, but the scope of protection of the present invention is not limited thereto. Any technician familiar with the technical field, within the technical scope disclosed by the present invention, who makes equivalent replacements or changes based on the technical solution and inventive concept of the present invention, should be covered by the scope of protection of the present invention.

Claims

1. A rapid diagnostic instrument for rabies virus, comprising a housing (1), characterized in that: One end of the shell (1) is fixedly connected to a moving handle (7), the edge of the other end of the shell (1) is rotatably connected to a rotating cover (2), the bottom of the shell (1) is fixedly connected to two support bars (8), the bottom of the inner wall of the shell (1) is fixedly connected to a base (25), the interior of the base (25) is rotatably connected to a rotating cylinder (24), one end of the rotating cylinder (24) is fixedly connected to a rotating disk, the upper surface of the shell (1) is rotatably connected to a first sealing cover (3), a second sealing cover (4) and a third sealing cover (6), the top of the inner wall of the shell (1) is fixedly connected to a vent cylinder (21), a plurality of filter plates are provided inside the vent cylinder (21), a plurality of air inlets (47) are provided on the upper surface of the vent cylinder (21), and a plurality of air inlet slits (20) are provided on the inner side surface of the vent cylinder (21); A collecting mechanism is installed at the bottom of the rotating cover (2), and the collecting mechanism is used to collect various experimental tools used for detecting rabies virus; A constant temperature mechanism is installed inside the housing (1), and the constant temperature mechanism is used to control the temperature inside the housing (1) and replace the air inside the housing (1); An automatic maintaining mechanism is installed inside the housing (1) near the constant temperature mechanism, and the automatic maintaining mechanism cooperates with the constant temperature mechanism to culture BHK cells.

2. A rapid diagnostic instrument for rabies virus according to claim 1, characterized in that: The collecting mechanism comprises a maintenance liquid tank (12) arranged inside the shell (1) and located at the bottom of the rotating cover (2); a cell cylinder (11), a virus cylinder (13), a maintenance liquid cylinder (14), a fluorescent liquid cylinder (15) and a test tube chamber (16) are respectively arranged at positions near the maintenance liquid tank (12) inside the shell (1); two pipettes (17) are arranged at positions near one side of the test tube chamber (16) inside the shell (1); temperature sensors (30) are respectively arranged on the lower surfaces of the first sealing cover (3), the second sealing cover (4) and the third sealing cover (6); pulling blocks (10) are respectively arranged on the upper surfaces of the first sealing cover (3), the second sealing cover (4) and the third sealing cover (6); and torsion springs are respectively arranged at the connection points between the first sealing cover (3), the second sealing cover (4) and the third sealing cover (6) and the shell (1).

3. A rapid diagnostic instrument for rabies virus according to claim 2, characterized in that: The constant temperature mechanism comprises an air pump (23) fixedly connected to the bottom of the inner wall of the housing (1), an auxiliary pipe (39) and an air outlet pipe (40) are respectively provided on one side of the air pump (23), a three-way valve (38) is provided at one end of the air outlet pipe (40), a heating pipe (33) is provided on one side of the three-way valve (38), a mounting shell (37) is fixedly connected to the top of the heating box (22), the heating pipe (33) is located inside the mounting shell (37), and an ultraviolet lamp (32) is provided at one end of the heating pipe (33).

4. A rapid diagnostic instrument for rabies virus according to claim 3, characterized in that: A connecting pipe is provided at the bottom of the three-way valve (38), a heating box (22) is provided at the bottom of the inner wall of the shell (1), a plurality of heating plates (34) are provided inside the heating box (22), a cleaning cylinder (35) is provided between the plurality of heating plates (34), a plurality of deceleration plates (36) are provided inside the cleaning cylinder (35), and one end of the connecting pipe is connected to one end of the cleaning cylinder (35).

5. A rapid diagnostic instrument for rabies virus according to claim 4, characterized in that: The other end of the cleaning cylinder (35) is provided with a discharge pipe (9), one end of the ultraviolet lamp (32) passes through the ventilation cylinder (21), a plurality of ventilation holes are provided on the outer wall of the rotating cylinder (24), a hexagonal groove is provided on the top of the rotating cylinder (24), and a rotating handle (29) is provided on the top of the hexagonal groove.

6. A rapid diagnostic instrument for rabies virus according to claim 5, characterized in that: The automatic maintenance mechanism comprises a micro pump (26) fixedly connected to the bottom of the inner wall of the housing (1), a liquid suction pipe (27) and a liquid discharge pipe (28) are respectively provided on one side of the micro pump (26), a connecting pipe (18) is provided at the bottom of the first sealing cover (3), a plurality of dripping pipes are provided at the bottom of the connecting pipe (18), and a solenoid valve is respectively provided at one end of each dripping pipe.

7. A rapid diagnostic instrument for rabies virus according to claim 6, characterized in that: A ninety-six-hole plate (19) is provided on the upper surface of the rotating disk at the bottom position of the connecting tube (18), and a plurality of culture holes are provided on the upper surface of the ninety-six-hole plate (19), the number of the culture holes and the number of the dripping tubes being the same, and one end of the drainage tube (28) is connected to the interior of the connecting tube (18).

8. A rapid diagnostic instrument for rabies virus according to claim 7, characterized in that: A plurality of locking mechanisms are installed on the upper surface of the housing (1) near the rotating cylinder (24), and the locking mechanisms are used to lock the first sealing cover (3), the second sealing cover (4) and the third sealing cover (6) to maintain the sealing inside the housing (1). The locking mechanisms include a plurality of clamping blocks (43) slidably connected to the upper surface of the housing (1) near the rotating cylinder (24), a first spring (42) is provided on one side of the clamping block (43), and an intermediate groove is provided inside the clamping block (43).

9. A rapid diagnostic instrument for rabies virus according to claim 8, characterized in that: A controller (5) is provided on the upper surface of the third sealing cover (6), and a button and a USB interface are provided on the upper surface of the controller (5). A camera (31) and an ultraviolet lamp (32) are provided on the bottom of the third sealing cover (6). A mounting rod (45) is slidably connected to the interior of the middle groove on the upper surface of the housing (1), a pressing button (41) is provided on the top of the mounting rod (45), a second spring (44) is provided on the bottom of the mounting rod (45), and a limiting block (46) is provided on the bottom of the pressing button (41).

10. The method for using a rapid diagnostic instrument for rabies virus according to claim 9, characterized in that: The following steps are involved: S1. Preparation: Rotate the first sealing cover (3) by ninety degrees, first place the ninety-six-well plate (19) on the rotating disk at the bottom of the first sealing cover (3), the upper surface of the rotating disk is provided with a plurality of limit blocks for fixing the ninety-six-well plate (19), the limit blocks are used to fix the position of the ninety-six-well plate (19), use a pipette (17) to drop the cell liquid inside the cell tube (11) into the plurality of culture wells on the upper surface of the ninety-six-well plate (19); S2. Culture: Take out multiple test tubes from the inside of the test tube warehouse (16), use a pipette (17) to dilute the rabies virus stock solution in the virus cylinder (13) and the cell culture maintenance solution in the maintenance solution cylinder (14) in proportion, and dilute them into ten levels of concentration. Since there are (96) culture wells on the upper surface of the ninety-six-well plate (19), the mixed solution of each concentration is dripped into eight culture wells in the vertical direction, and the twelve columns are dripped with the mixed solution of ten concentrations from left to right respectively. The eleventh column and the twelfth column are only dripped with the maintenance solution as a blank control group, which needs to be cultured for four to five days; S3, staining: by rotating the handle (29) to rotate the rotating cylinder (24) and the rotating disk, the ninety-six-well plate (19) is rotated from the bottom of the first sealing cover (3) to the bottom of the second sealing cover (4), the second sealing cover (4) is rotated to expose the upper surface of the ninety-six-well plate (19), and the fluorescent liquid in the fluorescent liquid cylinder (15) is dripped into the inside of the culture well using a pipette (17) to perform fluorescent staining on the BHK cells, and then allowed to stand for one hour; S4. Counting: The ninety-six-well plate (19) is rotated from the bottom of the second sealing cover (4) to the bottom of the third sealing cover (6). The ultraviolet lamp (32) emits ultraviolet light to make the part of the BHK cells infected with rabies virus emit fluorescence. The camera (31) takes pictures of the luminous cells in multiple culture wells on the upper surface of the ninety-six-well plate (19). The number of cells in the mixed solution of different concentrations is counted by image recognition technology, and the half infection dose of BHK cells is calculated by the Reed-Muench method.

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