A PCR detection device for rainbow shrimps virus

Abstract

This invention provides a PCR detection device for prawn iridovirus, belonging to the field of virus detection technology. It solves the technical problems of existing detection devices being easily contaminated by aerosols and unsuitable for rapid on-site detection. The device includes a shell with a touch screen display, a switch and discharge port on the rear, and a cover hinged to the top. Inside the shell, at the top, are a fluorescence detection component and several ultraviolet lamps. Inside the shell are a rechargeable battery, a detection box, and a main control board. The detection box has a rubber sealing gasket at the top and contains a pretreatment mechanism and a separation mechanism. The separation mechanism houses an amplification component. This invention automates sample processing and result analysis, significantly improving detection efficiency and consistency. It constructs multiple barriers to effectively prevent aerosol contamination and cross-contamination, enabling precise temperature control and detection, rapid amplification and sensitive signal capture, intelligent process coordination, real-time analysis and display of detection results, and high efficiency and reliability.

Description

Technical Field

[0001] This utility model belongs to the field of virus detection technology, and relates to a PCR detection device, particularly a PCR detection device for shrimp iridovirus. Background Technology

[0002] The iridovirus of freshwater shrimp is a double-stranded DNA virus. The virus particles are icosahedral and have an envelope. This virus is extremely harmful to the freshwater shrimp farming industry. After infection, it can cause symptoms such as atrophy of the hepatopancreas, lightening of color, empty intestines and stomach, cessation of feeding, and decreased vitality in shrimp. It spreads rapidly, has a high morbidity rate, and can cause large-scale mortality of freshwater shrimp in a short period of time, resulting in huge economic losses.

[0003] Currently, PCR technology has become the mainstream method for detecting iridovirus in freshwater shrimp due to its high sensitivity and specificity. Existing PCR testing procedures typically require multiple open workstations for nucleic acid extraction, reagent preparation, PCR amplification, and product analysis, involving numerous steps. Human operation can easily introduce aerosol contamination, leading to false positive results and severely impacting detection accuracy. Furthermore, it is generally performed in a laboratory, which is time-consuming and cannot meet the needs of rapid on-site testing in places such as aquaculture farms.

[0004] Therefore, we propose a PCR detection device for prawn iridovirus. By automating sample processing and result analysis, it significantly improves detection efficiency and consistency. By constructing multiple barriers, it effectively eliminates aerosol contamination and cross-contamination. Through precise temperature control and detection, it achieves rapid amplification and sensitive signal capture. The intelligent coordination process and real-time analysis of detection results simplify operation and ensure reliable results. Utility Model Content

[0005] The purpose of this invention is to address the aforementioned problems in existing technologies by proposing a PCR detection device for prawn iridovirus. The technical problem this invention aims to solve is: how to achieve rapid and accurate PCR detection of prawn iridovirus while avoiding aerosol contamination and cross-contamination.

[0006] The objective of this utility model can be achieved through the following technical solutions:

[0007] A PCR detection device for prawn iridovirus includes a housing with an air pump inside. A touchscreen display is located on the front bevel of the housing, and a switch and discharge port are located on the rear side. A cover plate is hinged to the top of the housing, and a handle is located on the cover plate. A fluorescence detection component and several ultraviolet lamps are located at the top inside the housing. From left to right, the interior of the housing contains a rechargeable battery, a detection box, and a main control board. A rubber sealing gasket is located at the top of the detection box, abutting against the top of the interior of the housing. From front to back, the interior of the detection box contains a pretreatment mechanism and a separation mechanism. The separation mechanism has an amplification component. The separation mechanism detects… The box is internally divided into a pretreatment chamber, a first disinfection channel, an amplification chamber, a second disinfection channel, and a detection chamber, arranged sequentially from front to back. Several ultraviolet lamps are located above the pretreatment chamber, the first disinfection channel, the amplification chamber, the second disinfection channel, and the detection chamber, respectively. An air pump pipe extends into the pretreatment chamber. The pretreatment mechanism is located inside the pretreatment chamber. The amplification component is located inside the amplification chamber. The fluorescence detection component is located inside the detection chamber. The discharge port is located in the middle of the rear side of the detection box. The main control board is electrically connected to the touch screen, the rechargeable battery, the fluorescence detection component, the pretreatment mechanism, the amplification component, the separating mechanism, the switch, and several ultraviolet lamps.

[0008] The working principle of this utility model is as follows: The operator turns on the switch, and the rechargeable battery powers the entire device; then, the handle is pulled to open the cover. At this time, the air pump starts and airflow is introduced into the pre-processing chamber through the pipe, forming an outward positive pressure airflow barrier at the inlet of the cover, effectively preventing external aerosols from entering the device and causing contamination. Then, the pre-processing mechanism moves to the inlet of the cover, and the operator places the nucleic acid pre-processing tube and nucleic acid carrier into the corresponding positions of the pre-processing mechanism. Then, the cover is closed, and the detection parameters are set through the touch screen. Then, the equipment is started.

[0009] The pretreatment unit shakes and heats the nucleic acid pretreatment tubes, then extracts iridovirus nucleic acid and injects it into the nucleic acid vector after standing. At the same time, the ultraviolet lamp above the first disinfection channel is turned on. After disinfection, the separation unit opens the transport channel between the first disinfection channel and the pretreatment chamber, and pushes the nucleic acid vector containing nucleic acid into the first disinfection channel. Then the transport channel is closed. At this time, the ultraviolet lamps above the pretreatment chamber and the amplification chamber are turned on simultaneously for disinfection. After disinfection, the separation unit opens the transport channel between the amplification chamber and the first disinfection channel, and transports the nucleic acid vector with the sample added to the amplification chamber. The amplification component moves to the top of the nucleic acid vector with the sample added to perform the PCR amplification reaction.

[0010] During the amplification reaction, the ultraviolet lamps above the disinfection channel 1, disinfection channel 2, and detection chamber are turned on sequentially for disinfection. The separation mechanism then opens the corresponding channels to sequentially transport the amplified nucleic acid vector to disinfection channel 2 and the detection chamber. Subsequently, fluorescence detection is performed inside the detection chamber by the fluorescence detection component, and the detection results are displayed on the touch screen in real time. After the detection is completed, the separation mechanism opens the rear channel of the detection chamber to discharge the tested nucleic acid vector from the discharge port of the outer shell.

[0011] The pretreatment mechanism includes a third electric lead screw arranged in the left-right direction and two first electric lead screws arranged symmetrically and vertically. A mounting plate is provided between the lead screw slides of the two first electric lead screws. The mounting plate is provided with a heating oscillator, a syringe holder, a third electric push rod, and a microfluidic chip. The microfluidic chip is provided with a sample dispensing port and is located behind the heating oscillator. A push plate is provided on the telescopic end of the third electric push rod and is located behind the microfluidic chip. The syringe holder is located on the left side of the heating oscillator. The third electric lead screw is fixedly installed inside the upper front side of the detection box. A second electric lead screw is detachably installed on the lead screw slide of the third electric lead screw, which is arranged in the front-rear direction. A vertically arranged precision injection pump is detachably installed on the lead screw slide of the second electric lead screw and is located above the heating oscillator.

[0012] Using the above structure, firstly, the synchronous movement of the two electric lead screw components drives the entire mounting plate and its components to move up and down to adjust to a suitable working height. Initially, the mounting plate rises, causing the heating oscillator to move to the inlet of the cover plate. Then, the operator places the nucleic acid pretreatment tube and the microfluidic chip onto the heating oscillator and the mounting plate, respectively. The microfluidic chip serves as the nucleic acid carrier, and the heating oscillator on the mounting plate is responsible for oscillating and heating the nucleic acid pretreatment tube, eluting the nucleic acid washing solution inside, and releasing the nucleic acid. During this process... During the process, the lead screw slide of the electric lead screw component three moves left and right, while the electric lead screw component two mounted on the slide is responsible for moving back and forth. The precision injection pump on the lead screw slide of the electric lead screw component two can perform aspiration and injection, and accurately move to the needle holder to replace or store the needle, move to the top of the heating oscillator for sampling, and accurately position the sample dispensing port of the microfluidic chip for nucleic acid transfer. After the nucleic acid is injected into the microfluidic chip, the electric push rod three on the mounting plate is activated, and the push plate on its telescopic end moves forward to push out the microfluidic chip that has completed the sample dispensing and send it to the next process.

[0013] The separation mechanism includes a conveyor and five vertically arranged partitions. The five partitions cooperate with the test box to divide the interior of the test box into a pretreatment chamber, a first disinfection channel, an amplification chamber, a second disinfection channel, and a test chamber arranged sequentially from front to back. The conveyor is located at the bottom inside the test box and is situated between the first partition on the front side and the fifth partition on the rear side. The five partitions have the same structure. The height of the first and fifth partitions is higher than that of the second, third, and fourth partitions. The width of the conveyor belt is equal to the width of the microfluidic chip. The partitions are all engaged inside the test box, and each partition has a sealing gasket at its lower end. The second, third, and fourth partitions abut against the conveyor, while the lower ends of the first and fifth partitions abut against the bottom inside the test box.

[0014] Using the above structure, the partitioning mechanism, through five vertically arranged partitions that cooperate with the inner wall of the detection box, sequentially divides the internal space into a pretreatment chamber, a sterilization channel one, an amplification chamber, a sterilization channel two, and a detection chamber, thus forming a sealed transport channel. The conveyor runs through the area between the first and fifth partitions, and its conveyor belt width is equal to the width of the microfluidic chip. It is responsible for carrying and accurately transporting the microfluidic chip sequentially through each chamber and channel. All partitions have sealing gaskets at their lower ends to ensure the airtightness between chambers. The lower ends of the second, third, and fourth partitions abut against the conveyor, allowing the microfluidic chip to pass through on the conveyor belt while achieving separation. The first and fifth partitions, due to their height, have their lower ends directly abut against the inner bottom of the detection box, effectively sealing both ends of the process channel. During operation, each partition opens or closes its lower channel sequentially under the command of the main control board. In conjunction with the intermittent movement of the conveyor, it guides the microfluidic chip to gradually complete the entire process from sample pretreatment to final detection, effectively preventing cross-contamination between chambers.

[0015] The lower end of each partition is provided with a sliding groove, the width of which is equal to the width of the conveyor belt. The front side of each partition is provided with a guide groove, which is located above and connected to the sliding groove. A lifting plate is slidably installed inside each sliding groove. A vertically installed electric push rod is fixed to the front side of each partition. A connecting plate is provided on the telescopic end of the electric push rod, which extends into the guide groove and is fixedly connected to the lifting plate.

[0016] With the above structure, each partition in the separation mechanism has a groove at its lower end, the width of which is the same as the width of the conveyor belt of the conveyor, to accommodate the sliding of the lifting plate. The telescopic end of the electric push rod is connected to the lifting plate in the groove through a connecting plate. The connecting plate can move up and down in the guide groove located above the groove. During operation, the main control board controls the extension and retraction of the electric push rod, driving the connecting plate to move along the guide groove, thereby driving the lifting plate to move up and down precisely in the groove. When the lifting plate descends to be flush with the surface of the conveyor belt, the channel closes, forming an effective seal. When the lifting plate rises to touch the top of the groove, the channel opens, allowing the microfluidic chip to pass through, thereby realizing the opening and closing control of each compartment and channel.

[0017] The amplification component includes a second electric push rod, which is fixedly positioned at the middle of the rear end face of the second partition. A semiconductor temperature control block is fixed below the telescopic end of the second electric push rod, and the specifications of the semiconductor temperature control block are matched with the microfluidic chip.

[0018] With the above structure, after the microfluidic chip is transported by the conveyor to the designated position in the amplification chamber, the telescopic end of the electric push rod two extends downward, driving the semiconductor temperature control block connected below to move precisely downward until it makes close contact with the corresponding area of ​​the microfluidic chip. Subsequently, under the control of the main control board, the semiconductor temperature control block executes a precise heating and cooling program to provide the temperature cycle necessary for PCR amplification of the reactants in the microfluidic chip. After the amplification reaction is completed, the electric push rod two retracts, driving the semiconductor temperature control block to rise and reset, separating it from the microfluidic chip so that it can be continued to be transported to the next process.

[0019] The fluorescence detection component includes an LED light source holder and a photomultiplier tube. The LED light source holder is fixedly installed at the top of the housing. The photomultiplier tube is installed on the LED light source holder, and the LED light source holder is located in the middle position between the fourth and fifth partitions. An optical lens and a filter plate are provided below the LED light source holder, with the filter plate located below the optical lens.

[0020] Using the above structure, when the amplified microfluidic chip is transported into the detection chamber and positioned between the fourth and fifth partitions, the LED light source emits light of a specific wavelength. The light passes sequentially through the optical lens and filter below it. The optical lens is responsible for focusing the light to precisely illuminate the detection area of ​​the microfluidic chip, exciting the fluorescent material inside to emit fluorescence. The filter is used to filter out stray light and excitation light, ensuring that only the target fluorescence signal can pass through. Subsequently, the fluorescence signal is captured by the photomultiplier tube above and converted into an electrical signal. After the signal is processed and analyzed by the main control board, the final detection result can be displayed on the touch screen.

[0021] Compared with existing technologies, this shrimp iridovirus PCR detection device has the following advantages:

[0022] 1. Through the coordinated operation of the pretreatment mechanism, the separation mechanism and the amplification component, the entire process from sample pretreatment, nucleic acid extraction, PCR amplification to fluorescence detection is automated, which significantly improves detection efficiency and consistency.

[0023] 2. Through multiple anti-pollution measures, a positive pressure airflow is generated by an air pump when the cover is opened. Ultraviolet lamps are installed above the pretreatment chamber, disinfection channel one, amplification chamber, disinfection channel two, and testing chamber. Combined with the sealed channel design of the separation mechanism, aerosol pollution and cross-contamination are avoided to the greatest extent.

[0024] 3. The semiconductor temperature control block of the amplification component is in close contact with the microfluidic chip under the drive of the electric push rod 2, ensuring the accuracy of the PCR amplification reaction temperature cycle; the fluorescence detection component achieves sensitive and accurate capture and analysis of fluorescence signals through the coordinated work of LED light source holder, optical lens, filter plate and photomultiplier tube.

[0025] 4. Parameters can be set intuitively and real-time results can be viewed through the touch screen. The main control board controls the entire process. The conveyor and the lifting plate that can be opened and closed precisely ensure continuous and stable sample flow. Finally, the tested chips are automatically discharged through the discharge port. While ensuring ease of operation, the reliability of the test results is also guaranteed. Attached Figure Description

[0026] Figure 1 This is a schematic diagram of the structure of this utility model.

[0027] Figure 2 This is a partially cutaway schematic diagram of this utility model.

[0028] Figure 3 This is a three-dimensional structural diagram of some components in this utility model.

[0029] Figure 4 This is a top view of some components of this utility model.

[0030] Figure 5 This is a partial cross-sectional structural diagram of some components in this utility model.

[0031] Figure 6 This is a schematic diagram of the separation mechanism in this utility model.

[0032] In the diagram: 1. Outer shell; 2. Touch screen; 3. Cover plate; 4. Handle; 5. Main control board; 6. Detection box; 7. Switch; 8. Rechargeable battery; 9. Separation mechanism; 10. Electric lead screw component one; 11. Electric push rod one; 12. Fluorescence detection component; 13. Partition plate; 14. Electric lead screw component two; 15. Precision syringe pump; 16. Electric lead screw component three; 17. Heating oscillator; 18. Needle holder; 19. Microfluidic chip; 20. Electric push rod two; 21. Semiconductor temperature control block; 22. Conveyor; 23. Electric push rod three; 24. Push plate; 25. Slide rail; 26. Lifting plate; 27. Connecting plate; 28. Guide groove. Detailed Implementation

[0033] The following are specific embodiments of the present invention, which are described in conjunction with the accompanying drawings. However, the present invention is not limited to these embodiments.

[0034] like Figures 1-6 As shown, this shrimp iridovirus PCR detection device includes a shell 1, an air pump inside the shell 1, a touch screen 2 on the front inclined surface of the shell 1, a switch 7 and a discharge port on the rear side of the shell 1, a cover plate 3 hinged to the upper end of the shell 1, a handle 4 on the cover plate 3, a fluorescence detection component 12 and several ultraviolet lamps inside the top of the shell 1, and a rechargeable battery 8, a detection box 6 and a main control board 5 arranged sequentially from left to right inside the shell 1, a rubber sealing gasket on the upper end of the detection box 6, which abuts against the top of the shell 1, and a pretreatment mechanism and a separation mechanism 9 arranged sequentially from front to back inside the detection box 6, with an amplification component on the separation mechanism 9. Mechanism 9 divides the interior of the test box 6 into a pretreatment chamber, a disinfection channel one, an amplification chamber, a disinfection channel two, and a test chamber arranged sequentially from front to back. Several ultraviolet lamps are located above the pretreatment chamber, disinfection channel one, amplification chamber, disinfection channel two, and test chamber, respectively. An air pump pipe extends into the pretreatment chamber. The pretreatment mechanism is located inside the pretreatment chamber. The amplification component is located inside the amplification chamber. The fluorescence detection component 12 is located inside the test chamber. The discharge port is located in the middle of the rear side of the test box 6. The main control board 5 is electrically connected to the touch screen 2, the rechargeable battery 8, the fluorescence detection component 12, the pretreatment mechanism, the amplification component, the separation mechanism 9, the switch 7, and several ultraviolet lamps, respectively.

[0035] In this embodiment, the operator turns on switch 7, and the rechargeable battery 8 powers the entire device. Then, the operator pulls handle 4 to open cover 3. At this time, the air pump starts and airflow is introduced into the pretreatment chamber through the pipe. A positive pressure airflow barrier is formed at the inlet of cover 3 to effectively prevent external aerosols from entering the device and causing contamination. Then, the pretreatment mechanism moves to the inlet of cover 3, and the operator places the nucleic acid pretreatment tube and nucleic acid carrier into the corresponding positions of the pretreatment mechanism. Then, cover 3 is closed, and the detection parameters are set through the touch screen 2. Then, the device is started.

[0036] The pretreatment unit shakes and heats the nucleic acid pretreatment tubes, then extracts iridovirus nucleic acid and injects it into the nucleic acid vector after standing. At the same time, the ultraviolet lamp above the first disinfection channel is turned on. After disinfection, the separation unit 9 opens the transport channel between the first disinfection channel and the pretreatment chamber, and pushes the nucleic acid vector containing nucleic acid into the first disinfection channel. Then the transport channel is closed. At this time, the ultraviolet lamps above the pretreatment chamber and the amplification chamber are turned on simultaneously for disinfection. After disinfection, the separation unit 9 opens the transport channel between the amplification chamber and the first disinfection channel, and transports the nucleic acid vector with the sample added to the amplification chamber. The amplification component moves to the top of the nucleic acid vector with the sample added to perform the PCR amplification reaction.

[0037] During the amplification reaction, the ultraviolet lamps above the disinfection channel 1, disinfection channel 2, and detection chamber are turned on in sequence for disinfection. The separation mechanism 9 then opens the corresponding channels to sequentially transport the amplified nucleic acid vector to disinfection channel 2 and the detection chamber. Subsequently, the fluorescence detection component 12 performs fluorescence detection inside the detection chamber, and the detection results are displayed on the touch screen 2 in real time. After the detection is completed, the separation mechanism 9 opens the rear channel of the detection chamber to discharge the tested nucleic acid vector from the discharge port of the outer shell 1.

[0038] The pretreatment mechanism includes a third electric lead screw 16 arranged in the left-right direction and two first electric lead screws 10 arranged symmetrically in the left and right directions and vertically. A mounting plate is provided between the lead screw slides of the two first electric lead screws 10. The mounting plate is provided with a heating oscillator 17, a syringe holder 18, a third electric push rod 23 and a microfluidic chip 19. The microfluidic chip 19 is provided with a sample dispensing port and is located behind the heating oscillator 17. A push plate 24 is provided on the telescopic end of the third electric push rod 23 and is located behind the microfluidic chip 19. The syringe holder 18 is located to the left of the heating oscillator 17. The third electric lead screw 16 is fixedly installed inside the upper front side of the detection box 6. A second electric lead screw 14 arranged in the front-rear direction is detachably provided on the lead screw slide of the third electric lead screw 16. A vertically arranged precision injection pump 15 is detachably provided on the lead screw slide of the second electric lead screw 14 and is located above the heating oscillator 17.

[0039] In this embodiment, firstly, the synchronous movement of the lead screw slides of the two electric lead screw components 10 drives the entire mounting plate and its components to move up and down to adjust to a suitable working height. Initially, the mounting plate rises, causing the heating oscillator 17 to move to the inlet of the cover plate 3. Subsequently, the operator places the nucleic acid pretreatment test tube and the microfluidic chip 19 on the heating oscillator 17 and the mounting plate, respectively. The microfluidic chip 19 serves as a nucleic acid carrier, and the heating oscillator 17 on the mounting plate is responsible for oscillating and heating the nucleic acid pretreatment test tube, eluting the nucleic acid washing solution inside, and releasing the nucleic acid. During this process, the electric... The lead screw slide of the moving lead screw component 3 16 moves left and right, while the electric lead screw component 2 14 mounted on the slide is responsible for moving back and forth. The precision injection pump 15 on the lead screw slide of the electric lead screw component 2 14 can draw and inject liquid, and move precisely to the needle holder 18 to replace or store the needle, move to the heating oscillator 17 to collect samples, and accurately position the sample dispensing port of the microfluidic chip 19 for nucleic acid transfer. After the nucleic acid is injected into the microfluidic chip 19, the electric push rod 3 23 on the mounting plate is activated, and the push plate 24 on its telescopic end moves forward to push out the microfluidic chip 19 that has completed the sample dispensing and send it to the next process.

[0040] The separating mechanism 9 includes a conveyor 22 and five vertically arranged partitions 13. The five partitions 13 cooperate with the test box 6 to divide the interior of the test box 6 into a pretreatment chamber, a disinfection channel one, an amplification chamber, a disinfection channel two, and a test chamber arranged sequentially from front to back. The conveyor 22 is located at the bottom inside the test box 6, and is positioned between the first partition 13 on the front side and the fifth partition 13 on the rear side. The five partitions 13 have the same structure. The height of the first partition 13 and the fifth partition 13 is higher than that of the second, third, and fourth partitions 13. The width of the conveyor belt of the conveyor 22 is equal to the width of the microfluidic chip 19. The partitions 13 are all engaged inside the test box 6. The lower end of each partition 13 is provided with a sealing gasket. The second, third, and fourth partitions 13 all abut against the conveyor 22. The lower ends of the first partition 13 and the fifth partition 13 abut against the bottom inside the test box 6.

[0041] In this embodiment, the partition mechanism 9, through five vertically arranged partitions 13 cooperating with the inner wall of the detection box 6, sequentially divides the internal space into a pretreatment chamber, a first disinfection channel, an amplification chamber, a second disinfection channel, and a detection chamber, thereby forming a sealed transport channel. The conveyor 22 passes through the area between the first and fifth partitions 13, and its conveyor belt width is equal to the width of the microfluidic chip 19, responsible for carrying and accurately transporting the microfluidic chip 19 sequentially through each chamber and channel. All partitions 13 have sealing gaskets at their lower ends to ensure the airtightness between chambers. The second... The lower ends of the first, third, and fourth partitions 13 abut against the conveyor 22, achieving separation while allowing the microfluidic chip 19 to pass through on the conveyor belt. The first and fifth partitions 13, due to their greater height, have their lower ends directly abut against the inner bottom of the detection box 6, thus sealing off both ends of the process channel. During operation, each partition 13 opens or closes its lower channel in sequence under the command of the main control board. In conjunction with the intermittent movement of the conveyor 22, the microfluidic chip 19 is guided to gradually complete the entire process from sample pretreatment to final detection, effectively preventing cross-contamination between the compartments.

[0042] Each partition 13 has a chute 25 at its lower end. The width of the chute 25 is equal to the width of the conveyor belt of the conveyor 22. Each partition 13 has a guide groove 28 on its front side. The guide groove 28 is located above the chute 25 and is connected to the chute 25. Each chute 25 has a lifting plate 26 that slides inside. Each partition 13 has a vertically installed electric push rod 11 fixed on its front side. Each electric push rod 11 has a connecting plate 27 on its telescopic end. The connecting plate 27 extends into the guide groove 28 and is fixedly connected to the lifting plate 26.

[0043] In this embodiment, each partition 13 in the partition mechanism 9 has a groove 25 at its lower end, the width of which is the same as the width of the conveyor belt of the conveyor 22, for accommodating the sliding of the lifting plate 26. The telescopic end of the electric push rod 11 is connected to the lifting plate 26 in the groove 25 through the connecting plate 27. The connecting plate 27 can move up and down in the guide groove 28 located above the groove 25. During operation, the main control board 5 controls the extension and retraction of the electric push rod 11, driving the connecting plate 27 to move along the guide groove 28, thereby driving the lifting plate 26 to move up and down precisely in the groove 25. When the lifting plate 26 descends to be flush with the surface of the conveyor belt of the conveyor 22, the channel is closed, forming an effective seal. When the lifting plate 26 rises to touch the top of the groove 25, the channel is opened, and the microfluidic chip 19 can pass through, thereby realizing the opening and closing control of each compartment and channel.

[0044] The amplification assembly includes a second electric push rod 20, which is fixedly positioned at the middle of the rear end face of the second partition 13. A semiconductor temperature control block 21 is fixed below the telescopic end of the second electric push rod 20, and the specifications of the semiconductor temperature control block 21 are matched with those of the microfluidic chip 19.

[0045] In this embodiment, after the microfluidic chip 19 is transported by the conveyor 22 to the designated position in the amplification chamber, the telescopic end of the electric push rod 20 extends downward, driving the semiconductor temperature control block 21 connected below to move precisely downward until it makes close contact with the corresponding area of ​​the microfluidic chip 19. Subsequently, under the control of the main control board 5, the semiconductor temperature control block 21 executes a precise heating and cooling program to provide the temperature cycle necessary for PCR amplification for the reactants in the microfluidic chip 19. After the amplification reaction is completed, the electric push rod 20 retracts, driving the semiconductor temperature control block 21 to rise and reset, separating it from the microfluidic chip 19 so that it can be transported to the next process.

[0046] The fluorescence detection component includes an LED light source holder and a photomultiplier tube. The LED light source holder is fixedly installed at the top inside the housing 1. The photomultiplier tube is installed on the LED light source holder, and the LED light source holder is located in the middle position between the fourth partition 13 and the fifth partition 13. An optical lens and a filter plate are provided below the LED light source holder, and the filter plate is located below the optical lens.

[0047] In this embodiment, when the amplified microfluidic chip 19 is transported into the detection chamber and positioned between the fourth and fifth partitions 13, the LED light source emits light of a specific wavelength. The light passes sequentially through the optical lens and filter below it. The optical lens is responsible for focusing the light to precisely illuminate the detection area of ​​the microfluidic chip 19, exciting the fluorescent material inside to emit fluorescence. The filter is used to filter out stray light and excitation light, ensuring that only the target fluorescence signal can pass through. Subsequently, the fluorescence signal is captured by the photomultiplier tube above and converted into an electrical signal. After the signal is processed and analyzed by the main control board 5, the final detection result can be displayed on the touch screen 2.

[0048] The working principle of this utility model is as follows: When the operator turns on the switch 7, the rechargeable battery 8 powers the entire device. Then, the operator pulls the handle 4 to open the cover 3. At this time, the air pump starts and airflow is introduced into the pre-processing chamber through the pipe, forming a positive pressure airflow barrier that outputs from the inlet of the cover 3 to prevent external aerosol contamination. Then, the pre-processing mechanism moves to the inlet of the cover 3. The operator places the nucleic acid pre-processing test tube and the microfluidic chip 19 on the heating oscillator 17 and the mounting plate, respectively. Then, the cover 3 is closed, and the detection parameters are set and the device is started through the touch screen 2.

[0049] The synchronous movement of the lead screw slides of the two electric lead screw components 10 drives the mounting plate to rise and fall. The heating oscillator 17 oscillates and heats the nucleic acid pretreatment tube to release nucleic acid. At the same time, the electric lead screw component 3 16, the electric lead screw component 2 14 and the precision injection pump 15 work together to drive the precision injection pump 15 to complete the actions of moving to the needle holder 18 to change the needle, moving to the top of the heating oscillator 17 to take samples and accurately positioning the microfluidic chip 19 sample dispensing port to transfer nucleic acid. After the nucleic acid injection is completed, the telescopic end of the electric push rod 3 23 drives the push plate 24 to push out the microfluidic chip 19 that has been sampled.

[0050] The conveyor 22 carries the microfluidic chip 19 through each compartment in sequence. The lifting plate 26 at the lower end of each partition 13 is precisely raised and lowered under the drive of the telescopic end of the electric push rod 11 to realize the opening and closing control of each compartment channel. Before the microfluidic chip 19 enters the first disinfection channel, the ultraviolet lamp above it is turned on for disinfection. Then the separation mechanism 9 opens the channel to send it into the first disinfection channel. At this time, the ultraviolet lamps above the pretreatment chamber and the amplification chamber are turned on simultaneously for disinfection.

[0051] Inside the amplification chamber, the amplification components begin to work. The telescopic end of the electric push rod 20 drives the semiconductor temperature control block 21 to move down and make close contact with the microfluidic chip 19. Under the control of the main control board 5, it performs precise temperature cycling to complete the PCR amplification reaction.

[0052] After amplification, the ultraviolet lamps above the second disinfection channel and the detection chamber are turned on in sequence for disinfection. After disinfection, the separation mechanism 9 opens the corresponding channel to transport the microfluidic chip 19 to the second disinfection channel. When the corresponding transport channel is opened, the microfluidic chip 19 is transported to the detection chamber. The fluorescence detection component 12 is then activated. The excitation light emitted by the LED light source holder is focused by the optical lens and filtered by the filter plate, and then accurately irradiates the detection area of ​​the microfluidic chip 19. The excited fluorescence signal is captured by the photomultiplier tube and converted into an electrical signal. After being processed and analyzed by the main control board 5, the detection result is displayed on the touch screen 2 in real time.

[0053] Finally, the separation mechanism 9 opens the rear channel of the testing chamber and discharges the tested microfluidic chip 19 from the discharge port of the outer shell 1, completing the entire testing process.

[0054] In summary, through the coordinated operation of the pretreatment mechanism, the separation mechanism 9, and the amplification component, the entire process from sample pretreatment, nucleic acid extraction, PCR amplification to fluorescence detection is automated, significantly improving detection efficiency and consistency.

[0055] Through multiple anti-pollution measures, a positive pressure airflow is generated by an air pump when the cover plate 3 is opened. Ultraviolet lamps are installed above the pretreatment chamber, disinfection channel one, amplification chamber, disinfection channel two and detection chamber. Combined with the sealed channel design of the separation mechanism 9, aerosol pollution and cross-contamination are avoided to the greatest extent.

[0056] The semiconductor temperature control block 21 of the amplification component is in close contact with the microfluidic chip 19 under the drive of the electric push rod 20, ensuring the accuracy of the PCR amplification reaction temperature cycle; the fluorescence detection component 12 achieves sensitive and accurate capture and analysis of fluorescence signals through the coordinated work of LED light source holder, optical lens, filter plate and photomultiplier tube.

[0057] The parameters can be set intuitively and the real-time results can be viewed through the touch screen 2. The main control board 5 controls the entire process in an overall manner. The conveyor 22 and the lifting plate 26 that can be opened and closed precisely ensure continuous and stable sample flow. Finally, the tested chips are automatically discharged through the discharge port. While ensuring ease of operation, the reliability of the test results is also ensured.

[0058] The specific embodiments described herein are merely illustrative examples illustrating the spirit of this utility model. Those skilled in the art to which this utility model pertains may make various modifications or additions to the described specific embodiments or use similar methods to substitute them, without departing from the spirit of this utility model or exceeding the scope defined by the appended claims.

Claims

1. A PCR detection device for prawn iridovirus, comprising a shell (1), characterized in that, The outer casing (1) is equipped with an air pump. A touch screen display (2) is provided on the front inclined surface of the outer casing (1). A switch (7) and a discharge port are provided on the rear side of the outer casing (1). A cover plate (3) is hinged to the upper end of the outer casing (1). A handle (4) is provided on the cover plate (3). A fluorescence detection component (12) and several ultraviolet lamps are provided on the top inside the outer casing (1). From left to right, the inner side of the outer casing (1) is equipped with a rechargeable battery (8), a detection box (6), and a main control board (5). A rubber sealing gasket is provided on the upper end of the detection box (6). The rubber sealing gasket abuts against the top inside the outer casing (1). From front to back, the inner side of the detection box (6) is equipped with a pretreatment mechanism and a separation mechanism (9). An amplification component is provided on the separation mechanism (9). The structure (9) divides the interior of the test box (6) into a pretreatment chamber, a disinfection channel one, an amplification chamber, a disinfection channel two, and a test chamber arranged sequentially from front to back. Several ultraviolet lamps are located above the pretreatment chamber, disinfection channel one, amplification chamber, disinfection channel two, and test chamber respectively. The air pump pipe extends into the interior of the pretreatment chamber. The pretreatment mechanism is located inside the pretreatment chamber. The amplification component is located inside the amplification chamber. The fluorescence detection component (12) is located inside the test chamber. The discharge port is located in the middle of the rear side of the test box (6). The main control board (5) is electrically connected to the touch screen (2), the rechargeable battery (8), the fluorescence detection component (12), the pretreatment mechanism, the amplification component, the separation mechanism (9), the switch (7), and several ultraviolet lamps respectively.

2. The PCR detection device for prawn iridovirus according to claim 1, characterized in that, The pretreatment mechanism includes a third electric lead screw (16) arranged in the left-right direction and two first electric lead screws (10) arranged symmetrically in the left and right directions and vertically. A mounting plate is provided between the lead screw slides of the two first electric lead screws (10). The mounting plate is provided with a heating oscillator (17), a syringe holder (18), a third electric push rod (23), and a microfluidic chip (19). The microfluidic chip (19) is provided with a sample dispensing port and is located behind the heating oscillator (17). A push plate is provided on the telescopic end of the third electric push rod (23). (24) The push plate (24) is located on the rear side of the microfluidic chip (19), the needle holder (18) is located on the left side of the heating oscillator (17), the electric lead screw three (16) is fixedly installed on the upper front side inside the detection box (6), the electric lead screw two (14) is detachably provided on the lead screw slide of the electric lead screw three (16) along the front and rear direction, the electric lead screw two (14) is detachably provided on the lead screw slide of the electric lead screw two (14) vertically arranged precision injection pump (15), the precision injection pump (15) is located above the heating oscillator (17).

3. The PCR detection device for prawn iridovirus according to claim 2, characterized in that, The partitioning mechanism (9) includes a conveyor (22) and five vertically arranged partitions (13). The five partitions (13) cooperate with the test box (6) to divide the interior of the test box (6) into a pretreatment chamber, a disinfection channel one, an amplification chamber, a disinfection channel two, and a test chamber arranged sequentially from front to back. The conveyor (22) is located at the bottom inside the test box (6), and the conveyor (22) is located between the first partition (13) on the front side and the fifth partition (13) on the rear side. The five partitions (13) have the same structure. The height of the partition (13) and the fifth partition (13) is higher than that of the second, third and fourth partitions (13). The width of the conveyor belt of the conveyor (22) is equal to the width of the microfluidic chip (19). The partitions (13) are all locked inside the detection box (6). The lower end of each partition (13) is provided with a sealing gasket. The second, third and fourth partitions (13) all abut against the conveyor (22). The lower ends of the first partition (13) and the fifth partition (13) abut against the inner bottom of the detection box (6).

4. The PCR detection device for prawn iridovirus according to claim 3, characterized in that, The lower end of each partition (13) is provided with a chute (25), the width of which is equal to the width of the conveyor belt of the conveyor (22). The front side of each partition (13) is provided with a guide groove (28), which is located above the chute (25) and connected to the chute (25). A lifting plate (26) is slidably provided inside the chute (25). A vertically arranged electric push rod (11) is fixed on the front side of each partition (13). A connecting plate (27) is provided on the telescopic end of the electric push rod (11), and the connecting plate (27) extends into the guide groove (28) and is fixedly connected to the lifting plate (26).

5. The PCR detection device for prawn iridovirus according to claim 4, characterized in that, The amplification component includes an electric push rod two (20), which is fixedly installed at the middle position of the rear end face of the second partition (13). A semiconductor temperature control block (21) is fixed below the telescopic end of the electric push rod two (20), and the specifications of the semiconductor temperature control block (21) are matched with the microfluidic chip (19).

6. The PCR detection device for prawn iridovirus according to claim 5, characterized in that, The fluorescence detection component includes an LED light source holder and a photomultiplier tube. The LED light source holder is fixedly installed at the top inside the housing (1). The photomultiplier tube is installed on the LED light source holder, and the LED light source holder is located in the middle position between the fourth partition (13) and the fifth partition (13). An optical lens and a filter plate are provided below the LED light source holder, and the filter plate is located below the optical lens.

Images