A device and method for detecting giant panda coronavirus particles

Abstract

The application relates to the technical field of biology, and particularly relates to a giant panda coronavirus particle detection device and method. The device comprises a bottom plate, a driving element is fixedly connected to the top of the bottom plate, a main gear is coaxially fixedly connected to the output end of the driving element, an extension rod is coaxially fixedly connected to the top of the main gear, a centrifugal assembly for centrifuging a to-be-detected sample is arranged at the other end of the extension rod, a first spring is coaxially fixedly connected to the top of the main gear, the other end of the first spring is fixedly connected to the bottom of the centrifugal assembly, a first rotating shaft is fixedly connected to the top of the centrifugal assembly, and a liquid supplementing assembly for adding a lysis solution to the centrifugal assembly is arranged at the top of the first rotating shaft. The centrifugal assembly, the liquid supplementing assembly and the piston assembly are synchronously driven by the main gear and the driven gear, integrated operation of sample centrifuging and automatic addition of the lysis solution is realized, manual intervention is reduced to avoid sample pollution, the centrifuging process is not interrupted, and the detection pretreatment period is shortened.

Description

Technical Field

[0001] This invention relates to the field of biotechnology, specifically to a device and method for detecting giant panda coronavirus particles. Background Technology

[0002] Giant pandas, a rare and endangered wild animal unique to my country, are a key species for global biodiversity conservation, and their population health and reproduction are directly related to the stability of the ecosystem. In recent years, coronavirus infection has become a significant threat to the health of giant pandas. These viruses easily cause inflammation of the respiratory and digestive tracts in giant pandas, which can lead to complications and even death in severe cases, posing a significant threat to the survival of both captive and wild giant panda populations. Therefore, rapid and accurate detection of giant panda coronavirus particles is a core technological requirement for timely disease prevention and control and for ensuring the health of giant pandas.

[0003] In existing technologies, such as the LAWSON SCIENTIFIC SCM-4K centrifuge-mixer, the core application of this equipment is sample pretreatment in the life sciences field. It is compatible with carriers such as 8-tube sets and 1.5 / 2mL centrifuge tubes, and can switch between three operating modes: centrifugation, mixing, and centrifugal mixing. The mixing intensity is divided into three levels: strong, medium, and gentle.

[0004] However, in practical use, this device only integrates centrifugation and mixing functions; the addition of lysis buffer still requires manual operation. The lysis buffer must be added to the sample tube by opening the cap during centrifugation and mixing, which not only breaks the operational process and prolongs the pretreatment cycle but also easily leads to sample exposure and contamination, affecting the integrity of viral nucleic acid and the accuracy of detection. Therefore, it is necessary to propose a device and method for detecting giant panda coronavirus particles. Summary of the Invention

[0005] To address the aforementioned issues, this invention provides a giant panda coronavirus particle detection device and method. This device and method use a main gear and a driven gear to synchronously drive a centrifugation assembly, a liquid replenishment assembly, and a piston assembly, thereby achieving integrated operation of sample centrifugation and automatic addition of lysis buffer. This reduces manual intervention to avoid sample contamination and avoids interrupting the centrifugation process, thus shortening the detection pretreatment cycle.

[0006] To achieve the above objectives, the technical solution of the present invention is as follows: A giant panda coronavirus particle detection device includes a base plate, a driving component fixedly connected to the top of the base plate, a main gear fixedly connected coaxially to the output end of the driving component, a telescopic rod fixedly connected coaxially to the top of the main gear, a centrifugation assembly for centrifuging the sample to be tested at the other end of the telescopic rod, a first spring fixedly connected coaxially to the top of the main gear, the other end of the first spring fixedly connected to the bottom of the centrifugation assembly, a first rotating shaft fixedly connected to the top of the centrifugation assembly, a replenishment assembly for adding lysis buffer to the centrifugation assembly at the top of the first rotating shaft, a replenishment assembly housing and a housing cover, and a baffle slidably fitted on the inner side wall of the housing. The first rotating shaft, at its end furthest from the centrifugal assembly, passes through the box body, baffle, and lid, and is detachably connected to these components. An air inlet is located at the top of the lid, and several liquid outlets are fixedly connected to the bottom of the box body. Each liquid outlet has a pressure valve fixedly connected to its other end. A main gear meshes with a driven gear, which is coaxially and fixedly connected to a second rotating shaft. One end of the second rotating shaft passes through the driven gear and rotates with the base plate. The other end of the second rotating shaft is equipped with a transmission assembly for transmitting power. A piston assembly for inflating the air inlet is located at the top of the base plate. An adjustment assembly for adjusting the output of the piston assembly is located between the power input end of the piston assembly and the push rod. The gas output end of the piston assembly is connected to the air inlet.

[0007] The technical principles of the above solution are as follows: The sample is loaded into a centrifuge tube, and the lysis buffer is filled into the box below the baffle. The replenishment assembly is moved downward along the first rotating shaft, and the outlet head enters the centrifuge tube and is fixed. The increased weight of the rotating platform due to the added sample compresses the first spring downward. The adjustment assembly adjusts the gas output of the piston assembly according to the downward stroke of the rotating platform. The drive unit is activated, and its output end drives the rotating platform to rotate via the main gear and telescopic rod, thus centrifuging the sample in the centrifuge tube. Simultaneously, the first rotating shaft drives the replenishment assembly to rotate synchronously with the rotating platform. The main gear drives the driven gear through the transmission assembly and adjustment assembly to drive the piston assembly. The piston assembly fills the air inlet with gas, which enters the box and pushes the baffle downward, increasing the pressure of the lysis buffer. This causes the lysis buffer to pass through the pressure valve, and then through the outlet head into the corresponding centrifuge tube. Under the action of the lysis buffer and the rotating platform, the sample is lysed in the centrifuge tube.

[0008] The above approach has the following beneficial effects: 1. This invention integrates sample centrifugation and automatic lysis buffer addition functions into one unit, realizing automated operation from sample loading to lysis completion. It eliminates the need for manual step-by-step opening of the cap to add lysis buffer, solving the problems of cumbersome operation and process interruption of existing equipment, and shortening the sample pretreatment cycle.

[0009] 2. This invention uses the change in the weight of the rotating stage during sample loading to trigger the adjustment component, thereby achieving dynamic adaptive adjustment of the amount of lysis buffer added. The more samples are loaded, the more lysis buffer is added, avoiding inconsistencies in the amount added each time when added manually. This ensures that samples of different loading volumes and types can be fully lysed, improving the consistency and reliability of the test results.

[0010] 3. During the centrifugation and lysis process of this invention, the lysis buffer is directly injected into the rotating centrifuge tube through the outlet head, without exposing the sample throughout the process, effectively reducing the impact of external contamination on viral nucleic acid. At the same time, centrifugation and lysis are carried out simultaneously, and the lysis buffer is fully mixed with the sample under the action of centrifugal force, further improving the lysis efficiency of viral particles and providing a high-quality sample matrix for subsequent nucleic acid purification and amplification.

[0011] Furthermore, the centrifuge assembly includes a rotating platform coaxially and fixedly connected to the telescopic rod and the top of the first spring. Several centrifuge tubes are embedded and installed on the rotating platform, and each centrifuge tube is located on the same axis as its adjacent liquid outlet. The top of the rotating platform is coaxially and fixedly connected to the first rotating shaft.

[0012] Beneficial effects: The embedded installation design of the centrifuge tubes ensures stable loading during rotation. The centrifuge tubes correspond one-to-one with the liquid outlets, and the rotating table and the liquid replenishment assembly rotate coaxially and synchronously, ensuring that the lysate can be accurately injected into the centrifuge tubes during rotation.

[0013] Furthermore, the transmission assembly includes a rotating rod fixedly connected to the top end of the second rotating shaft, a transmission rod rotatably engaged at the other end of the rotating rod, a first slider rotatably engaged at the other end of the transmission rod, and a push rod fixedly connected to the other end of the first slider; it also includes a first slide rail fixedly connected to the top of the base plate, and the first slide rail slidably engaged with the first slider.

[0014] Beneficial effects: By cooperating with the rotating rod, transmission rod, first slider and first slide rail, the rotational motion of the gear is converted into the linear reciprocating motion of the push rod. The transmission structure is simple and efficient, ensuring that the piston assembly can continuously and stably obtain power and ensuring the continuity of pyrolysis fluid addition.

[0015] Furthermore, the piston assembly includes a piston cylinder and a piston rod. The piston rod extends through the side wall of the piston cylinder into the piston cylinder and slides within the piston cylinder. The other end of the piston cylinder has an outlet and an intake port. An exhaust check valve is fixedly connected to the outlet, and the gas flow direction in the exhaust check valve is from inside the piston cylinder to outside the piston cylinder. The outlet is fixedly connected to the intake port. An intake check valve is fixedly connected to the intake port, and the gas flow direction in the intake check valve is from outside the piston cylinder to inside the piston cylinder. A piston plate is fixedly connected to one end of the piston rod located inside the piston cylinder. The piston plate slides within the inner side wall of the piston cylinder. A third spring is fixedly connected to the side of the piston plate away from the piston rod. The other end of the third spring is fixedly connected to the end of the piston cylinder away from the piston rod. A piston seat is fixedly connected to the bottom of the piston cylinder, and the bottom of the piston seat is fixedly connected to the base plate.

[0016] Beneficial effects: The exhaust check valve and intake check valve ensure unidirectional gas flow and prevent backflow from affecting pressure stability; the third spring enables the piston plate to automatically reset without additional driving force; the piston seat provides support for the piston cylinder, enhancing stability and preventing vibration and displacement during operation.

[0017] Furthermore, the adjustment assembly includes a second slide rail, a second slider slidably fitted onto the second slide rail, a stop block fixedly connected to the bottom of the second slider, a first groove formed on one side wall of the stop block, a third slider slidably fitted onto the first groove, the third slider being hinged to the end of the piston rod away from the piston plate, and the side wall of the stop block away from the first groove abutting against the push rod; several fourth sliders are fixedly connected to the outer wall of the second slide rail, several third slide rails are fixedly connected to the top of the base plate, each fourth slider slidably fitted onto its adjacent third slide rail, and a second spring is fixedly connected between each fourth slider and the base plate; a transmission plate is fixedly connected to one side of the second slide rail, a second annular groove is formed on the bottom of the rotary table, and the other end of the transmission plate slidably fits into the second groove.

[0018] Beneficial effects: The weight change of the rotary table is converted into the lifting and lowering motion of the second slide rail by the transmission plate, which causes the stop to rise and fall, thereby changing the driving distance of the piston plate and realizing the dynamic adjustment of the inflation volume of the piston assembly. It can adapt to different sample loading volumes without manual intervention.

[0019] Furthermore, a ball is fixedly connected to the end of the push rod that abuts against the stop block.

[0020] Beneficial effects: The ball changes the contact between the push rod and the stop from a surface contact to a point contact, reducing wear during relative sliding. At the same time, the ball can adapt to the inclined contact surface of the stop, avoiding jamming, ensuring smooth power transmission, and extending the service life of transmission components.

[0021] Furthermore, sealing rings are fixedly connected to the sliding joints between the baffle and the inner side wall of the box and the first rotating shaft.

[0022] Beneficial effects: The sealing ring enhances the sealing between the baffle, the box and the first rotating shaft, preventing gas leakage from causing insufficient pressure of the pyrolysis liquid, while also avoiding contamination of the pyrolysis liquid and preventing waste caused by leakage of the pyrolysis liquid.

[0023] Furthermore, the main gear is an incomplete gear, the second rotating shaft is in unidirectional rotational engagement with the base plate, and the slave gear can be replaced with a reduction gear set.

[0024] Beneficial effects: The incomplete gear allows the piston assembly to be aerated only during specific time periods, enabling intermittent addition of the lysis fluid and avoiding excessive addition frequency that could affect the lysis effect; unidirectional rotation prevents the stop from being pushed back by the third spring when the driven gear loses driving force in the toothless section of the main gear, which would cause the driven gear to reverse and interrupt the piston pushing process; the reduction gear set allows for flexible adjustment of the transmission ratio to adapt to the lysis rate requirements of different samples, improving the adaptability and practicality of the equipment.

[0025] Furthermore, a method for detecting giant panda coronavirus particles includes the following steps: Step 1, Sample Collection and Preprocessing: For wild or captive giant pandas, collect one of the following: fresh fecal samples, pharyngeal swab samples, or tissue samples; add 3-5 mL of sterile saline to the fecal sample and shake to homogenize for 3 minutes; shake the pharyngeal swab thoroughly for 1 minute to wash away the virus; cut the tissue sample into small pieces and homogenize until there are no obvious particles to complete the sample preprocessing. Step 2, Loading Configuration: Load the pretreated samples into centrifuge tubes, add lysis buffer to the inside of the container, install the baffle above the lysis buffer surface, and then close the lid. Move the entire replenishment assembly along the first rotating shaft so that the outlet head extends into the centrifuge tube. Fix the container and lid to the first rotating shaft. According to the sample loading volume, the rotating stage moves downward by the corresponding stroke, causing the baffle to move down. The piston assembly increases the amount of gas output each time, and the amount of lysis buffer added to the centrifuge tube each time increases accordingly. Step 3, sample centrifugation and lysis: Start the drive unit, the rotary table starts to rotate and centrifuges the sample in the centrifuge tube, and the first rotating shaft drives the liquid replenishment component to rotate synchronously with the rotary table. At the same time, the main gear drives the slave gear, and drives the transmission component to inflate the air inlet through the piston assembly at fixed time intervals, so that the baffle squeezes the lysis liquid. The lysis liquid flows out from the liquid outlet through the pressure valve and enters the centrifuge tube, thus lysing the sample in conjunction with the centrifugation motion. Step 4, Nucleic Acid Purification: After lysis is complete, turn off the equipment and wait for the rotating stage to come to a standstill before removing the centrifuge tubes to purify the nucleic acids in the mixture. Step 5, Nucleic Acid Amplification: Amplify the purified nucleic acid and collect fluorescence data; Step 6, Result Interpretation: Analyze the fluorescence curve. A positive result is defined as a curve with a clear growth curve and an inflection point cycle number ≤ 35. A negative result is defined as a curve with no growth curve or no clear inflection point. Estimate the viral load based on the fluorescence peak value.

[0026] Beneficial effects: This method designs preprocessing schemes for different scenarios and sample types of giant pandas to ensure sample applicability; the automatic centrifugation and lysis by this device and the standardized operation process reduce human error, making the test results more reliable and providing accurate data support for disease prevention and control.

[0027] Furthermore, in step three, different tooth ratios of the main gear and the driven gear are selected to change the time interval between each addition of pyrolysis solution.

[0028] Beneficial effects: By adjusting the ratio of the number of teeth in the master and slave gears, the interval for adding lysis buffer can be flexibly controlled to adapt to the lysis characteristics of different samples. For tissue samples that are difficult to lyse, the interval can be shortened and the frequency of adding lysis buffer can be increased. For pharyngeal swab samples that are easy to lyse, the interval can be extended to avoid reagent waste and further improve the flexibility and detection specificity of the equipment. Attached Figure Description

[0029] Figure 1 This is an isometric view of an embodiment of the giant panda coronavirus particle detection device of the present invention.

[0030] Figure 2 This is a cross-sectional view of an embodiment of the giant panda coronavirus particle detection device of the present invention.

[0031] Figure 3 This is a bottom view of an embodiment of the giant panda coronavirus particle detection device of the present invention.

[0032] Figure 4 This is a step diagram of an embodiment of the giant panda coronavirus particle detection method of the present invention.

[0033] The reference numerals in the accompanying drawings of the instruction manual include: 1. Base plate; 2. Drive component; 3. Main gear; 4. Telescopic rod; 5. First spring; 6. Rotary table; 7. Centrifuge tube; 8. First shaft; 9. Box body; 10. Box cover; 11. Baffle; 12. Air inlet; 13. Liquid outlet; 14. Second shaft; 15. Driven gear; 16. Rotating rod; 17. Transmission rod; 18. First slider; 19. First slide rail; 20. Push rod; 21. Second slider; 22. Second slide rail; 23. Stop block; 24. Third slider; 25. First slide groove; 26. Fourth slider; 27. Third slide rail; 28. Second spring; 29. ​​Piston rod; 30. Piston cylinder; 31. Piston plate; 32. Piston seat; 33. Air outlet; 34. Air intake; 35. Transmission plate; 36. Second slide groove; 37. Third spring. Detailed Implementation

[0034] The technical solution of the present invention will now be clearly and completely described with reference to the accompanying drawings. Obviously, the described embodiments are only some, not all, of the embodiments of the present invention. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0035] In the description of this invention, it should be noted that the terms "center," "upper," "lower," "left," "right," "vertical," "horizontal," "inner," and "outer," etc., indicate the orientation or positional relationship based on the orientation or positional relationship shown in the accompanying drawings. They are used only for the convenience of describing the invention and for simplifying the description, and do not indicate or imply that the device or element referred to must have a specific orientation, or be constructed and operated in a specific orientation. Therefore, they should not be construed as limitations on the invention. Furthermore, the terms "first," "second," and "third" are used for descriptive purposes only and should not be construed as indicating or implying relative importance.

[0036] In the description of this invention, it should be noted that, unless otherwise explicitly specified and limited, the terms "installation," "connection," and "linking" should be interpreted broadly. For example, they can refer to a fixed connection, a detachable connection, or an integral connection; they can refer to a mechanical connection or an electrical connection; they can refer to a direct connection or an indirect connection through an intermediate medium; and they can refer to the internal connection of two components. Those skilled in the art can understand the specific meaning of the above terms in this invention based on the specific circumstances.

[0037] The following detailed description illustrates the specific implementation method: Example 1: As attached Figure 1 and Figure 2 As shown: A giant panda coronavirus particle detection device includes a base plate 1, a drive component 2 bolted to the top of the base plate 1, a main gear 3 coaxially bolted to the output end of the drive component 2, a telescopic rod 4 coaxially bolted to the top of the main gear 3, and a centrifugation component for centrifuging the sample to be tested at the other end of the telescopic rod 4.

[0038] A first spring 5 is coaxially bonded to the top of the main gear 3. The other end of the first spring 5 is welded to the bottom of the centrifuge assembly. A first rotating shaft 8 is bolted to the top of the centrifuge assembly. A replenishment component for adding lysis solution to the centrifuge assembly is provided on the top of the first rotating shaft 8.

[0039] The liquid replenishment assembly includes a box body 9 and a box cover 10; a baffle 11 is slidably fitted on the inner side wall of the box body 9; the end of the first rotating shaft 8 away from the centrifugal assembly passes through the box body 9, the baffle 11 and the box cover 10 and is detachably connected to the box body 9, the baffle 11 and the box cover 10; an air inlet 12 is opened on the top of the box cover 10; a number of liquid outlets 13 are fixedly connected to the bottom of the box body 9; and a pressure valve is fixedly connected to the other end of each liquid outlet 13.

[0040] The centrifuge assembly includes a rotating platform 6 coaxially welded to the top of the telescopic rod 4 and the first spring 5. Several centrifuge tubes 7 are embedded in the rotating platform 6. All centrifuge tubes 7 are located on the same axis as their adjacent liquid outlet heads 13. The top of the rotating platform 6 is coaxially bolted to the first rotating shaft 8.

[0041] Specifically, the sample to be tested is placed in centrifuge tube 7, and drive unit 2 is activated. The output end of drive unit 2 drives the main gear 3 to rotate, the main gear 3 drives the telescopic rod 4 to rotate, the telescopic rod 4 drives the rotating table 6 to rotate, and the rotating table 6 and the main gear 3 drive the first spring 5 to rotate synchronously. When the rotating table 6 rotates, it drives the centrifuge tube 7 to rotate, thereby centrifuging the sample in the centrifuge tube 7; at the same time, the first rotating shaft 8 drives the box body 9, the box cover 10 and the liquid outlet 13 to rotate synchronously. In this embodiment, drive unit 2 is a motor.

[0042] like Figure 1 and Figure 2 As shown, the main gear 3 meshes with the driven gear 15, and the driven gear 15 is coaxially bolted to a second rotating shaft 14. One end of the second rotating shaft 14 passes through the driven gear 15 and is rotatably engaged with the base plate 1. The other end of the second rotating shaft 14 is provided with a transmission component for transmitting power.

[0043] The transmission assembly includes a rotating rod 16 bolted to the top of the second rotating shaft 14, a transmission rod 17 rotatably engaged at the other end of the rotating rod 16, a first slider 18 rotatably engaged at the other end of the transmission rod 17, and a push rod 20 fixedly connected to the other end of the first slider 18. It also includes a first slide rail 19 bolted to the top of the base plate 1, the first slide rail 19 slidingly engaging with the first slider 18.

[0044] Specifically, when the main gear 3 rotates, it drives the driven gear 15 to rotate, the driven gear 15 drives the second rotating shaft 14 to rotate, the second rotating shaft 14 drives the rotating rod 16 to rotate, the other end of the rotating rod 16 drives the transmission rod 17 to move, the other end of the transmission rod 17 drives the first slider 18 to slide back and forth on the first slide rail 19, and the first slider 18 drives the push rod 20 to move back and forth.

[0045] like Figure 1 and Figure 2 As shown, the top of the base plate 1 is provided with a piston assembly for inflating the air inlet 12. The piston assembly includes a piston cylinder 30 and a piston rod 29. The piston rod 29 extends through the side wall of the piston cylinder 30 into the piston cylinder 30 and slides with the piston cylinder 30. The other end of the piston cylinder 30 has an air outlet 33 and an air intake 34. An exhaust check valve is fixedly connected in the air outlet 33. The gas flow direction in the exhaust check valve is from the inside of the piston cylinder 30 to the outside of the piston cylinder 30. The air outlet 33 is fixedly connected to the air inlet 12. An intake check valve is fixedly connected in the air intake 34. The gas flow direction in the intake check valve is from the outside of the piston cylinder 30 to the inside of the piston cylinder 30. A piston plate 31 is fixedly connected to one end of the piston rod 29 inside the piston cylinder 30. The piston plate 31 slides with the inner wall of the piston cylinder 30. A third spring 37 is welded to the side of the piston plate 31 away from the piston rod 29. The other end of the third spring 37 is welded to the end of the piston cylinder 30 away from the piston rod 29. A piston seat 32 is welded to the bottom of the piston cylinder 30. The bottom of the piston seat 32 is bolted to the base plate 1.

[0046] Specifically, when the piston rod 29 is subjected to external force, it pushes the piston plate 31 to compress the third spring 37. The piston plate 31 discharges the gas in the piston cylinder 30 through the air outlet 33 and enters the air inlet 12. When the piston rod 29 loses external force, the third spring 37 returns to its original state and pushes the piston plate 31 to reset. At this time, the external gas is drawn into the piston cylinder 30 through the air intake 34.

[0047] like Figure 1 and Figure 2 and Figure 3 As shown, an adjustment component for adjusting the output of the piston assembly is provided between the power input end of the piston assembly and the push rod 20. The adjustment component includes a second slide rail 22, a second slider 21 slidably fitted on the second slide rail 22, a stop block 23 bolted to the bottom of the second slider 21, a first groove 25 on one side wall of the stop block 23, a third slider 24 slidably fitted on the first groove 25, the third slider 24 being hinged to the end of the piston rod 29 away from the piston plate 31, and the side wall of the stop block 23 away from the first groove 25 abutting against the push rod 20; several fourth sliders 26 are bolted to the outer wall of the second slide rail 22; several third slide rails 27 are bolted to the top of the base plate 1, and each fourth slider 26 is slidably fitted with its adjacent third slide rail 27, and a second spring 28 is welded between each fourth slider 26 and the base plate 1; a transmission plate 35 is bolted to one side of the second slide rail 22, and a second annular groove 36 is opened at the bottom of the rotary table 6, with the other end of the transmission plate 35 slidably fitted with the second groove 36.

[0048] Specifically, such as Figure 2 As shown, when the rotary table 6 rotates, the left end of the transmission plate 35 can slide along the second slide groove 36; the stop block 23 can drive the second slider 21 to slide back and forth along the second slide rail 22, the second slide rail 22 can drive the fourth slider 26 to move up and down along the third slide rail 27, and the second spring 28 applies an upward thrust to the fourth slider 26.

[0049] After the centrifuge tube 7 is loaded with the sample, the rotating table 6 becomes heavier and compresses the first spring 5 downwards. The transmission plate 35 descends with the rotating table 6, thereby driving the second slide rail 22 to descend. The second slider 21 drives the stop 23 to descend with the second slide rail 22. The third slider 24 slides upwards relative to the first slide groove 25, and the stop 23 can reciprocate along the second slide rail 22 via the second slider 21. With the maximum distance to the right of the push rod 20 abutting on the left side of the stop 23 remaining unchanged, and the length of the piston rod 29 remaining unchanged, the push rod 20 pushes the stop 23 to move to the right because the stop 23 is narrower at the bottom and wider at the top. When the piston rod 29 and piston plate 31 are pushed to the right, the piston plate 31 moves a greater distance, thereby increasing the amount of gas pushed in a single operation and increasing the amount of lysate released in a single operation. Therefore, the amount of lysate released in a single operation can be automatically adjusted according to the weight of the loaded sample. When the push rod 20 moves to the left and the stop block 23 loses its thrust, the third spring 37 pushes the piston plate 31 to reset, and pushes the stop block 23 to reset through the piston rod 29. This ensures that when the piston rod 29 advances a greater distance, it can still reset to the initial position before the next push, thereby ensuring an effective increase in the amount of gas pushed.

[0050] This invention reduces operational complexity and minimizes process interruptions by incorporating a transmission assembly that simultaneously drives the centrifugal assembly via the main gear 3, which in turn drives the replenishment assembly to automatically replenish the centrifuge tube 7 with lysis buffer. Furthermore, by setting up an adjustment assembly, the single-stroke advance distance of the piston rod is automatically increased when the rotary table 6 becomes heavier and moves downwards, thereby automatically increasing the amount of lysis buffer added per stroke.

[0051] Example 2: As attached Figure 1 As shown, the difference from Embodiment 1 is that a ball is welded to the end of the push rod 20 that abuts against the stop block 23. The main gear 3 is an incomplete gear, the second rotating shaft 14 is in unidirectional rotational engagement with the base plate 1, and the slave gear 15 can be replaced with a reduction gear set. Sealing rings are bonded to the sliding engagement points of the baffle 11 with the inner wall of the box 9 and the first rotating shaft 8.

[0052] The push rod 20 moves the ball. When the ball contacts and pushes the stop 23, the ball can adapt to the inclined contact surface, reducing wear caused by mismatch in the contact surface. The main gear 3 intermittently drives the reduction gear set to rotate, extending the time of driving the transmission components, thereby reducing the frequency of adding pyrolysis fluid. The sealing ring improves the sealing performance of the baffle 11, thereby better separating the upper and lower chambers of the baffle 11.

[0053] Example 3: As attached Figure 4 As shown, the difference from Example 2 is that a method for detecting giant panda coronavirus particles includes the following steps: Step 1, Sample Collection and Preprocessing: For wild or captive giant pandas, collect one of the following: fresh fecal samples, pharyngeal swab samples, or tissue samples. In this example, fresh fecal samples are selected. Add 3-5 mL of sterile physiological saline to the fecal sample and shake to homogenize for 3 minutes. Shake the pharyngeal swab thoroughly for 1 minute to wash away the virus. Cut the tissue sample into small pieces and homogenize until there are no obvious particles. This completes the sample preprocessing.

[0054] Step 2, Loading Configuration: Load the pretreated samples into centrifuge tubes 7, add lysis buffer to the inside of the container 9, install baffle 11 above the lysis buffer surface, and then cover with the container lid 10. Move the entire replenishment assembly along the first rotating shaft 8 so that the outlet head 13 extends into the centrifuge tube 7, and fix the container 9 and the container lid 10 on the first rotating shaft 8. According to the sample loading volume, the rotating stage 6 moves downward by the corresponding stroke, causing the baffle 23 to move down, increasing the amount of gas output by the piston assembly each time, and thus increasing the amount of lysis buffer added to the centrifuge tube 7 each time.

[0055] Step 3, sample centrifugation and lysis: Start the drive unit 2, the rotary table 6 starts to rotate and centrifuges the sample in the centrifuge tube 7, and the first rotating shaft 8 drives the liquid replenishment component to rotate synchronously with the rotary table 6. At the same time, the main gear 3 drives the slave gear 15 to drive the transmission component to inflate the air inlet 12 through the piston assembly at fixed time intervals, so that the baffle 11 squeezes the lysis liquid. The lysis liquid flows out from the liquid outlet 13 through the pressure valve and enters the centrifuge tube 7, thus lysing the sample in conjunction with the centrifugation motion.

[0056] Step 4, Nucleic Acid Purification: After lysis, turn off the equipment and wait for the rotating stage 6 to come to a standstill before removing the centrifuge tube 7 to purify the nucleic acid in the mixture. In this embodiment, the magnetic bead method is used for purification.

[0057] Step 5, Nucleic Acid Amplification: The purified nucleic acid is amplified, and fluorescence data is collected. In this embodiment, a real-time PCR instrument is used for nucleic acid amplification.

[0058] Step 6, Result Interpretation: Analyze the fluorescence curve. A positive result is defined as a curve with a clear growth rate and an inflection point cycle number ≤ 35. A negative result is defined as a curve with no growth rate or no clear inflection point. Estimate the viral load based on the fluorescence peak value.

[0059] In step three, different tooth ratios are used for the primary gear 3 and the driven gear 15 to change the time interval between each addition of lysis fluid. In this embodiment, different tooth ratios include 12:1 and 16:1, corresponding to time intervals of 12 minutes and 20 minutes, respectively.

[0060] Obviously, the above embodiments are merely illustrative examples for clear explanation and are not intended to limit the implementation. Those skilled in the art will recognize that other variations or modifications can be made based on the above description. It is neither necessary nor possible to exhaustively list all possible implementations here. However, obvious variations or modifications derived therefrom are still within the scope of protection of this invention.

Claims

1. A giant panda coronavirus particle detection device, characterized in that, The system includes a base plate (1), a drive unit (2) fixedly connected to the top of the base plate (1), a main gear (3) fixedly connected to the output end of the drive unit (2) on the same axis, a telescopic rod (4) fixedly connected to the top of the main gear (3) on the same axis, a centrifugation assembly for centrifuging the sample to be tested provided at the other end of the telescopic rod (4), a first spring (5) fixedly connected to the top of the main gear (3) on the same axis, the other end of the first spring (5) fixedly connected to the bottom of the centrifugation assembly, a first rotating shaft (8) fixedly connected to the top of the centrifugation assembly, and a device for centrifuging the sample to be tested provided at the top of the first rotating shaft (8). A replenishment assembly for adding lysis buffer to a centrifuge assembly includes a replenishment assembly box (9) and a box cover (10). A baffle (11) is slidably fitted on the inner side wall of the box (9). The end of the first rotating shaft (8) away from the centrifuge assembly passes through the box (9), the baffle (11), and the box cover (10) and is detachably connected to the box (9), the baffle (11), and the box cover (10). An air inlet (12) is opened at the top of the box cover (10). Several liquid outlets (13) are fixedly connected to the bottom of the box (9). A pressure valve is fixedly connected to the other end of each liquid outlet (13). The main gear (3) meshes with the driven gear (15), and the driven gear (15) is coaxially fixedly connected to the second rotating shaft (14). One end of the second rotating shaft (14) passes through the driven gear (15) and rotates with the base plate (1). The other end of the second rotating shaft (14) is provided with a transmission component for transmitting power. The bottom plate (1) is provided with a piston assembly for inflating the air inlet (12) at the top. An adjustment assembly for adjusting the output of the piston assembly is provided between the power input end of the piston assembly and the push rod (20). The gas output end of the piston assembly is connected to the air inlet (12).

2. The giant panda coronavirus particle detection device according to claim 1, characterized in that, The centrifuge assembly includes a rotating platform (6) that is coaxially fixedly connected to the top of the telescopic rod (4) and the first spring (5). Several centrifuge tubes (7) are embedded in the rotating platform (6). All centrifuge tubes (7) are located on the same axis as their adjacent liquid outlets (13). The top of the rotating platform (6) is coaxially fixedly connected to the first rotating shaft (8).

3. The giant panda coronavirus particle detection device according to claim 2, characterized in that, The transmission assembly includes a rotating rod (16) fixedly connected to the top of the second rotating shaft (14), a transmission rod (17) rotatably connected to the other end of the rotating rod (16), a first slider (18) rotatably connected to the other end of the transmission rod (17), and a push rod (20) fixedly connected to the other end of the first slider (18); it also includes a first slide rail (19) fixedly connected to the top of the base plate (1), and the first slide rail (19) and the first slider (18) slidably connected.

4. The giant panda coronavirus particle detection device according to claim 3, characterized in that, The piston assembly includes a piston cylinder (30) and a piston rod (29). The piston rod (29) extends through the side wall of the piston cylinder (30) into the piston cylinder (30) and slides with the piston cylinder (30). The other end of the piston cylinder (30) has an outlet (33) and an intake (34). An exhaust check valve is fixedly connected inside the outlet (33). The gas flow direction inside the exhaust check valve is from inside the piston cylinder (30) to outside the piston cylinder (30). The outlet (33) is fixedly connected to the inlet (12). An intake check valve is fixedly connected inside the intake (34). The gas flow direction inside the intake check valve is from inside the piston cylinder (30) to outside the piston cylinder (30). The flow direction is from outside the piston cylinder (30) to inside the piston cylinder (30); the piston plate (31) is fixedly connected to one end of the piston rod (29) inside the piston cylinder (30), the piston plate (31) slides with the inner wall of the piston cylinder (30), the side of the piston plate (31) away from the piston rod (29) is fixedly connected to a third spring (37), the other end of the third spring (37) is fixedly connected to the end of the piston cylinder (30) away from the piston rod (29), the bottom of the piston cylinder (30) is fixedly connected to a piston seat (32), and the bottom of the piston seat (32) is fixedly connected to the bottom plate (1).

5. The giant panda coronavirus particle detection device according to claim 4, characterized in that, The adjustment assembly includes a second slide rail (22), a second slider (21) which is slidably fitted to the second slide rail (22), a stop block (23) which is fixedly connected to the bottom of the second slider block (21), a first groove (25) which is opened on one side wall of the stop block (23), a third slider (24) which is slidably fitted to the first groove (25), the third slider (24) which is hinged to the end of the piston rod (29) away from the piston plate (31), and the side wall of the stop block (23) away from the first groove (25) abuts against the push rod (20); the second slide rail (22) Several fourth sliders (26) are fixedly connected to the outer wall, and several third slide rails (27) are fixedly connected to the top of the base plate (1). The fourth sliders (26) slide in cooperation with their adjacent third slide rails (27). A second spring (28) is fixedly connected between the fourth sliders (26) and the base plate (1). A transmission plate (35) is fixedly connected to one side of the second slide rail (22). A second slide groove (36) in the shape of a ring is opened at the bottom of the rotary table (6). The other end of the transmission plate (35) slides in cooperation with the second slide groove (36).

6. The giant panda coronavirus particle detection device according to claim 5, characterized in that, A ball is fixedly connected to the end of the push rod (20) that abuts against the stop block (23).

7. The giant panda coronavirus particle detection device according to claim 6, characterized in that, A sealing ring is fixedly connected to the sliding fit of the baffle (11) with the inner wall of the box (9) and the first rotating shaft (8).

8. The giant panda coronavirus particle detection device according to claim 7, characterized in that, The main gear (3) is an incomplete gear, the second rotating shaft (14) is in unidirectional rotational engagement with the base plate (1), and the slave gear (15) can be replaced with a reduction gear set.

9. A method for detecting giant panda coronavirus particles, characterized in that, Includes the following steps: Step 1, Sample Collection and Preprocessing: For wild or captive giant pandas, collect one of the following: fresh fecal samples, pharyngeal swab samples, or tissue samples; add 3-5 mL of sterile saline to the fecal sample and shake to homogenize for 3 minutes; shake the pharyngeal swab thoroughly for 1 minute to wash away the virus; cut the tissue sample into small pieces and homogenize until there are no obvious particles to complete the sample preprocessing. Step 2, loading configuration: Load the pretreated samples into centrifuge tubes (7), add lysis buffer into the box body (9), install baffle (11) above the lysis buffer surface, and then cover the box with lid (10); move the entire replenishment assembly along the first rotating shaft (8) so that the liquid outlet (13) extends into the centrifuge tube (7), and fix the box body (9) and lid (10) on the first rotating shaft (8); according to the sample loading amount, the rotating table (6) moves downward by the corresponding stroke, so that the baffle (23) moves down, the amount of gas output by the piston assembly increases each time, and the amount of lysis buffer added to the centrifuge tube (7) each time increases accordingly; Step 3, sample centrifugation and lysis: Start the drive unit (2), the rotary table (6) starts to rotate and centrifuges the sample in the centrifuge tube (7), and the first rotating shaft (8) drives the liquid replenishment component to rotate synchronously with the rotary table (6). At the same time, the main gear (3) drives the slave gear (15) to drive the transmission component to inflate the air inlet (12) through the piston assembly at fixed time intervals, so that the baffle (11) squeezes the lysis liquid. The lysis liquid flows out from the liquid outlet (13) through the pressure valve and enters the centrifuge tube (7), thus lysing the sample in conjunction with the centrifugation motion. Step 4, nucleic acid purification: After lysis is completed, turn off the equipment and wait for the rotating stage (6) to come to rest before taking out the centrifuge tube (7) to purify the nucleic acid in the mixture; Step 5, Nucleic Acid Amplification: Amplify the purified nucleic acid and collect fluorescence data; Step 6, Result Interpretation: Analyze the fluorescence curve. A positive result is defined as a curve with a clear growth curve and an inflection point cycle number ≤ 35. A negative result is defined as a curve with no growth curve or no clear inflection point. Estimate the viral load based on the fluorescence peak value.

10. The method for detecting giant panda coronavirus particles according to claim 9, characterized in that, In step three, different tooth ratios of the main gear (3) and the driven gear (15) are selected to change the time interval between each addition of pyrolysis solution.

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