Compact structure digital PCR instrument and control method thereof

Abstract

The application provides a compact digital PCR instrument and a control method thereof. The PCR instrument comprises an integrated module and a rotating mechanism. The integrated module comprises a first workbench and a second workbench. The first workbench is provided with a chip placing groove. The second workbench is provided with a droplet generating module and a PCR amplification module. When the second workbench is in a working state, the droplet generating module is pressed and connected to a chip oil hole and a chip gas-liquid hole of an integrated micro-droplet chip to form a micro-droplet in the integrated micro-droplet chip. The reaction chamber of the integrated micro-droplet chip is in a temperature control cavity of the PCR amplification module. The rotating mechanism is used for driving the two workbenches to synchronously turn 180° when the second workbench is in the working state. The application has the advantages of compact structure, small instrument volume, high portability and low manufacturing cost.

Description

TECHNICAL FIELD

[0001] The application belongs to the technical field of digital PCR instrument device design, and particularly relates to a compact digital PCR instrument and a control method thereof. BACKGROUND

[0002] Digital PCR is the latest quantitative technology, and is a method of absolute quantification based on single-molecule PCR counting nucleic acid quantification. The method mainly uses microfluidic or microdroplet methods in the current hot research field of analytical chemistry to disperse a large amount of diluted nucleic acid solution into microreactors or microdroplets of a biochip. The number of nucleic acid templates in each reactor is less than or equal to 1. After PCR cycles, a reactor with one nucleic acid molecule template will give a fluorescent signal, and a reactor without a template will have no fluorescent signal. According to the relative proportion and the volume of the reactor, the number of template copies in the sample can be accurately calculated.

[0003] The existing digital PCR system is often complex in structure, large in size and high in cost. It not only occupies more valuable space in the laboratory, but also affects the popularization of digital PCR technology to some extent due to the high cost of the instrument. In addition, nucleic acid amplification is a key link in digital PCR technology, and its amplification efficiency directly affects the accuracy of the final detection result. The amplification efficiency of nucleic acid is mainly affected by the accuracy and consistency of temperature control. Since the microdroplet sample needs to be heated by a microfluidic chip in the digital PCR technology, the heat transfer efficiency is limited by the chip structure and the heat transfer performance of the material. A certain constant temperature time is needed to make the temperature of the microdroplets uniform, which puts higher requirements on the temperature uniformity of the temperature control module.

[0004] Patent application No. 202111381197.4 discloses an integrated digital PCR instrument and a control method thereof. The instrument has high automation and integration, which can improve the efficiency of detection and analysis and reduce labor costs. In the technical solution, a mechanical hand (i.e. a scheduling mechanism) is used to schedule the chips between the microdroplet generation module, the turnover module, the PCR amplification module, the detection module and the waste tank. Multiple chips work simultaneously to form a pipeline working mode. The chips sequentially complete the four links of droplet generation, chip turnover, droplet amplification and droplet fluorescence detection. In the technical solution, the mechanical hand needs to be scheduled in multiple modules, which results in an overall structure that is not compact enough, a relatively complex structure design, a large size, poor portability and relatively high manufacturing cost. SUMMARY

[0005] Therefore, the present application aims to provide a compact digital PCR instrument and a control method thereof, which effectively overcomes the shortcomings of the prior art, such as complex structure design, large instrument size, poor compactness, poor portability, and high manufacturing cost.

[0006] To solve the above problems, the present application provides a compact digital PCR instrument, which comprises an integrated module and a rotating mechanism. The integrated module comprises a first work platform and a second work platform. The first work platform has a plurality of chip placement grooves for positioning and placing an integrated micro-droplet chip. The second work platform is arranged in parallel with the first work platform and has a working state close to the first work platform and a chip taking and placing state away from the first work platform. The second work platform has a droplet generation module and a PCR amplification module. When the second work platform is in the working state, the droplet generation module and the chip oil hole and chip gas-liquid hole of the integrated micro-droplet chip are respectively pressed and connected to form micro-droplets in the integrated micro-droplet chip. The reaction chamber of the integrated micro-droplet chip is in the temperature control cavity of the PCR amplification module. The rotating mechanism is used to drive the first work platform and the second work platform to synchronously turn 180° when the second work platform is in the working state.

[0007] In some embodiments, the integrated module further comprises left and right spaced vertical plates. The length ends of the first work platform are pivotally arranged on the top end positions of the two vertical plates. The bottom end positions of the two vertical plates are connected by a connecting plate. The rotating mechanism comprises a rotary motor and a transmission assembly. The rotary motor is fixedly connected to one of the vertical plates. The rotary motor is drivingly connected to one end of the first work platform through the transmission assembly to drive the first work platform to rotate by a predetermined angle.

[0008] In some embodiments, the compact digital PCR instrument further comprises a device bottom plate and a longitudinal movement assembly. The longitudinal movement assembly comprises a longitudinal movement guide rail and a longitudinal movement motor. The bottom end of the integrated module is slidingly connected between the longitudinal movement guide rail and the device bottom plate. The longitudinal movement motor can drive the integrated module to reciprocatingly move linearly along the guide direction of the longitudinal movement guide rail. The guide direction of the longitudinal movement guide rail is perpendicular to the length direction of the first work platform.

[0009] In some embodiments, the compact digital PCR instrument further comprises a fluorescence detection module on the first side of the integrated module and a detection compression module on the second side of the integrated module, the first side and the second side being one of the top side and the bottom side of the first work platform, the fluorescence detection module being capable of being controlled to be lifted away from or close to the first work platform to achieve position correspondence with the fluorescence detection area of the integrated microdroplet chip and perform fluorescence detection, the detection compression module being capable of being controlled to be lifted away from or close to the second work platform to achieve compression docking with the first hole and the second hole in the droplet generation module, so that the detection oil in the detection oil circuit enters the reaction chamber to make the microdroplets after amplification float to the fluorescence detection area and enter the sample loading cavity of the integrated microdroplet chip.

[0010] In some embodiments, the detection compression module comprises an air cavity and a compression assembly connected to the air cavity, the air cavity being used to provide negative pressure for the droplet generation module and positive pressure for the detection compression module; and / or the fluorescence detection module and the detection compression module are capable of being controlled to move to be capable of corresponding to the position of each integrated microdroplet chip.

[0011] In some embodiments, the compression assembly comprises a detection compression block and a moving block, the detection compression block being in sliding connection with the moving block and being capable of being driven to be lifted by a first compression motor, the moving block being in sliding connection with the air cavity and being capable of being driven to move along the length direction of the first work platform by a second compression motor; preferably, the moving block is provided with a light source to provide a light field for a camera of the fluorescence detection module.

[0012] In some embodiments, the first work platform and the second work platform are each vertically provided with a guide slide rod at the corresponding corner area, the guide slide rod is sleeved with a first return spring, the first return spring is clamped between the first work platform and the second work platform, and further comprises a pressing structure, the pressing structure is capable of overcoming the elastic force of the first return spring under the action of a pressing force to switch the second work platform from the chip taking and placing state to the working state.

[0013] In some embodiments, the droplet generation module comprises a generation compression block, the length extension direction of the generation compression block is parallel to the width direction of the second work platform, and the length two ends of the generation compression block are connected with the second work platform via connecting lugs, the first hole and the second hole are formed on the generation compression block, and the droplet generation module further comprises a generation cover plate, the generation cover plate has a top surface for sealing the chip oil hole and the chip air-liquid hole when generating microdroplets and achieving compression communication of the air circuit on the generation cover plate with the chip air-liquid hole.

[0014] The application also provides a control method of the compact digital PCR instrument.

[0015] Placing the integrated micro-droplet chip loaded with sample liquid into the chip placing groove;

[0016] Switching the second operation platform from the chip taking and placing state to the working state;

[0017] Controlling the droplet generation module to generate micro-droplets in the integrated micro-droplet chip and store the micro-droplets in the reaction chamber of the integrated micro-droplet chip;

[0018] After the micro-droplets are generated, the first operation platform and the second operation platform are synchronously flipped by 180°, and the PCR amplification module is controlled to amplify the micro-droplets in the reaction chamber;

[0019] After the amplification is completed, the detection compression module and the fluorescence detection module are both moved towards the integrated micro-droplet chip, the detection compression module is controlled to drive the amplified micro-droplets to enter the sample adding cavity of the integrated micro-droplet chip from the reaction chamber through the fluorescence detection area of the integrated micro-droplet chip after the detection compression module and the droplet generation module have the first generation hole and the second generation hole, and the fluorescence detection module is controlled to perform fluorescence detection at the fluorescence detection area.

[0020] In some embodiments, the control method further comprises:

[0021] During the operation of the PCR amplification module, the first operation platform and the second operation platform are synchronously reciprocally swung; and / or,

[0022] After the fluorescence detection is completed, the first operation platform and the second operation platform are synchronously rotated by 180° in the reverse direction, and then the second operation platform is switched from the working state to the chip taking and placing state.

[0023] The compact digital PCR instrument and the control method thereof provided by the application integrate the droplet generation module and the PCR amplification module on the second operation platform, the second operation platform has a working state and a chip taking and placing state and can be switched between the two states, the first operation platform and the second operation platform are synchronously flipped by the rotation driving of the rotation mechanism, the integrated micro-droplet chip does not need to be switched between different operation modules or flipped by the scheduling mechanism in the prior art, the structure of the digital PCR instrument is more compact, the volume of the instrument is smaller, the portability is stronger, and the manufacturing cost is relatively reduced. BRIEF DESCRIPTION OF DRAWINGS

[0024] Figure 1 This is a schematic diagram of the structure of a compact digital PCR instrument according to an embodiment of the present invention (the overall structure omits parts such as the outer shell).

[0025] Figure 2 for Figure 1 A three-dimensional structural diagram of the integrated microdroplet chip in the image;

[0026] Figure 3 for Figure 1 A three-dimensional structural diagram of the integrated module (including the rotating mechanism);

[0027] Figure 4 yes Figure 3 A magnified view of a section at point A in the middle;

[0028] Figure 5 for Figure 1 A schematic diagram of the three-dimensional structure of the droplet generation module in the image;

[0029] Figure 6 This is a schematic diagram of the three-dimensional structure of a PCR amplification module;

[0030] Figure 7 This is a schematic diagram of the three-dimensional structure after the flipping mechanism is assembled.

[0031] Figure 8 This is a three-dimensional structural diagram of the fluorescence detection module and the detection crimping module assembled inside the instrument.

[0032] Figure 9 for Figure 8 A three-dimensional structural diagram of the detection crimping module in the image;

[0033] Figure 10 for Figure 8 A three-dimensional structural diagram of the fluorescence detection module in the image;

[0034] Figure 11 A schematic diagram of the instrument's three-dimensional structure before microdroplet generation;

[0035] Figure 12 A schematic diagram of the instrument's three-dimensional structure in the microdroplet generation state;

[0036] Figure 13 A schematic diagram of the instrument's three-dimensional structure during PCR amplification at 180° in the flipped-over state;

[0037] Figure 14 This is a schematic diagram of the local three-dimensional structure under fluorescence detection.

[0038] The reference numerals in the attached figures are as follows:

[0039] 1, first work platform; 11, vertical plate; 12, connecting plate; 121, guide slide rod; 122, first reset spring; 123, pressing rod; 124, roller; 125, traction rope; 126, linear bearing; 13, chip placement groove; 131, guide spring; 2, second work platform; 3, droplet generation module; 31, generation first hole; 32, generation second hole; 33, generation pressing block; 34, generation cover plate; 35, electromagnet; 36, rotating shaft; 37, second reset spring; 38, guide rod; 39, third reset spring; 310, cover plate gasket; 4, PCR amplification module; 41, semiconductor heating element; 42, heat insulation plate; 43, heat conduction block; 44, first heat dissipation block; 47, third heat dissipation block; 48, second heat dissipation block; 49, heat conduction pipe; 5, rotating mechanism; 51, rotary motor; 52, transmission assembly; 6, fluorescence detection module; 61, objective lens; 62, optical path assembly; 63, lifting motor; 64, translation motor; 7, detection pressure connection module; 70, detection oil circuit; 71, detection pressure connection block; 72, moving block; 73, first pressure connection motor; 74, second pressure connection motor; 75, first gasket; 76, air cavity; 77, light source; 100, integrated micro-droplet chip; 101, chip oil hole; 102, chip gas-liquid hole; 103, reaction chamber; 104, fluorescence detection area; 105, sample addition cavity; 201, device bottom plate; 2021, longitudinal movement guide rail; 2022, longitudinal movement motor. DETAILED DESCRIPTION

[0040] CONJUNCTION WITH Figures 1 to 14As shown, according to the embodiment of the present application, a compact digital PCR instrument is provided, which comprises an integrated module (not labeled in the figure) and a rotating mechanism 5 (not labeled in the figure), the integrated module comprises a first work platform 1 and a second work platform 2, the first work platform 1 is provided with a plurality of chip placement grooves 13 for positioning and placing an integrated micro-droplet chip 100, it can be understood that each chip placement groove 13 positions and places an integrated micro-droplet chip 100, the second work platform 2 is arranged in parallel with the first work platform 1 and has a working state close to the first work platform 1 and a chip taking and placing state away from the first work platform 1, the second work platform 2 is provided with a droplet generation module 3 and a PCR amplification module 4, when the second work platform 2 is in the working state, the droplet generation module 3 and the chip oil hole 101 and the chip gas-liquid hole 102 of the integrated micro-droplet chip 100 are respectively pressed and connected to form micro-droplets in the integrated micro-droplet chip 100, the formed micro-droplets will be stored in the reaction chamber 103 of the integrated micro-droplet chip 100, and the reaction chamber 103 of the integrated micro-droplet chip 100 is in the temperature control cavity (not shown, not labeled) of the PCR amplification module 4, the rotating mechanism 5 is used to drive the first work platform 1 and the second work platform 2 to synchronously overturn 180° when the second work platform 2 is in the working state, so that the reaction chamber 103 is upside down, at this time, it is ensured that the micro-droplets stored in the reaction chamber 103 can be at the bottom of the reaction chamber 103 under the action of gravity, and the temperature control cavity of the PCR amplification module 4 can more efficiently control the temperature of the micro-droplets (i.e. the reaction solution).

[0041] In the technical solution, the droplet generation module 3 and the PCR amplification module 4 are integrated on the second work platform 2, the second work platform 2 has a working state and a chip taking and placing state and can switch between the two states, and the first work platform 1 and the second work platform 2 are synchronously overturned by the rotation driving of the rotating mechanism 5, without using the scheduling mechanism in the prior art to switch the integrated micro-droplet chip 100 between different work modules or overturn it, so that the structure of the digital PCR instrument is more compact, the instrument volume is smaller, thereby having stronger portability, and the manufacturing cost is relatively reduced.

[0042] Specifically referring to Figure 7 As shown, the shape of the aforementioned chip placement groove 13 matches the shape of the bottom surface of the integrated micro-droplet chip 100, as a preferred embodiment, the side of the chip placement groove 13 away from the operation side of the operator is provided with a corresponding guide spring 131, so that the position of the chip is more reliable when the chip is placed, and when the chip is taken out, the end of the chip can be forced to pop out of the chip placement groove 13, thereby facilitating the taking out of the chip.

[0043] In some embodiments, the integrated module further comprises two vertically spaced upright plates 11, the first work platform 1 is pivotally (e.g. through a rolling pivot connection) arranged at the top ends of the two upright plates 11, and the bottom ends of the two upright plates 11 are connected by a connecting plate 12 to improve the structural reliability between the two upright plates 11, the rotating mechanism 5 comprises a rotary motor 51 and a transmission assembly 52, the rotary motor 51 is fixedly connected to one of the upright plates 11, and the rotary motor 51 is drivingly connected to one end of the first work platform 1 through the transmission assembly 52 to drive the first work platform 1 to rotate by a preset angle, in a specific embodiment, the rotary motor 51 and the transmission assembly 52 are realized by a rotary driving assembly on the market.

[0044] In the technical solution, the rotating mechanism 5 can drive the first work platform 1 to flip up and down by 180°, so that the top surface of the integrated microdroplet chip 100 faces upward to generate microdroplets, and after the microdroplet generation is completed, the first work platform 1 and the second work platform 2 are flipped up and down synchronously by rotation, so that the top surface of the integrated microdroplet chip 100 faces downward to realize that the microdroplets in the reaction chamber 103 can be at the bottom of the reaction chamber 103 under the action of gravity, thereby ensuring the reliable temperature control of the PCR amplification module 4 on the microdroplets in the reaction chamber 103, and the rotating mechanism 5 can also drive the first work platform 1 to swing back and forth during the amplification process, i.e. to rotate by the aforementioned preset angle (generally ±10°), so that the uniformity of the temperature field is accelerated by fluid convection, the heat transfer efficiency and temperature uniformity in the microdroplet sample are improved, and the temperature uniformity requirement of the temperature control module itself is also reduced, thereby improving the efficiency of the amplification link.

[0045] In some embodiments, referring to Figure 8 The compact digital PCR instrument further comprises a device bottom plate 201 and a longitudinal movement assembly (not labeled in the figure), the longitudinal movement assembly comprises a longitudinal movement guide rail 2021 and a longitudinal movement motor 2022, the bottom end of the integrated module is slidingly connected between the longitudinal movement guide rail 2021 and the device bottom plate 201, and the longitudinal movement motor 2022 can drive the integrated module to reciprocatingly move linearly along the guide direction of the longitudinal movement guide rail 2021, and the guide direction of the longitudinal movement guide rail 2021 is a horizontal direction perpendicular to the length direction of the first work platform 1.

[0046] In the technical solution, the longitudinal moving assembly can drive the integrated module to adjust the position in the horizontal longitudinal direction of the instrument, so that the integrated module adjusts the corresponding position with the corresponding processing module (for example, the subsequent detection and crimping module 7 and the fluorescence detection module 6) in different process flows, simplifies the structural design of the processing module, for example, the displacement control of the processing module in the longitudinal direction is not needed, that is, the control is simplified.

[0047] Further referring to Figure 8 As shown in the figure, in some embodiments, the compact digital PCR instrument further comprises a fluorescence detection module 6 on the first side of the integrated module and a detection and crimping module 7 on the second side of the integrated module, the first side and the second side are one of the top side and the bottom side of the first workbench 1, and in a specific embodiment, the first side is the top side of the first workbench 1 and the second side is the bottom side of the first workbench 1. Figure 8 In the orientation shown as a reference, the first side, that is, the upper side of the integrated module, and the second side, that is, the lower side of the integrated module, the fluorescence detection module 6 can be controlled to be lifted to move away from or close to the first workbench 1 to realize the position correspondence with the fluorescence detection area 104 of the integrated micro-droplet chip 100 and perform fluorescence detection, and the detection and crimping module 7 can be controlled to be lifted to move close to or away from the second workbench 2 to realize the pressure tight connection with the generation first hole 31 and the generation second hole 32 in the droplet generation module 3, so that the detection oil in the detection oil path 70 (which can be specifically understood as a detection oil bottle) can enter the reaction chamber 103 to make the micro-droplets after amplification reaction float to the fluorescence detection area 104 and enter the sample adding cavity 105 of the integrated micro-droplet chip 100, and the fluorescence detection module 6 and the detection and crimping module 7 can be controlled to move to correspond to each integrated micro-droplet chip 100.

[0048] In the technical solution, by arranging the fluorescence detection module 6 and the detection and crimping module 7 on the upper and lower sides of the integrated module, the PCR instrument of the application can integrate the fluorescence detection function of the micro-droplet, and realize the functional combination of the instrument.

[0049] Specifically referring to Figure 9 As shown in the figure, in some embodiments, the detection and crimping module 7 comprises an air cavity 76 and a crimping assembly (not labeled in the figure) connected to the air cavity 76, the air cavity 76 is used to provide negative pressure for the droplet generation module 3 and provide positive pressure for the detection and crimping module 7, wherein the negative pressure is provided for the micro-droplet generation process during the operation of the droplet generation module 3, and the positive pressure is provided for the supply of the detection oil during the operation of the fluorescence detection module 6.

[0050] In the technical solution, the air cavity 76 can be used as a pressure container of the instrument and also as an assembly carrier of the crimping assembly, which can further simplify the structural design of the instrument and improve the compactness of the instrument.

[0051] Continuing to refer to Figure 9 As shown, in some embodiments, the crimping assembly comprises a detection crimping block 71 and a moving block 72, the detection crimping block 71 is in sliding connection with the moving block 72 and can be driven to lift by a first crimping motor 73, the moving block 72 is in sliding connection with the air cavity 76 and can be driven to move laterally along the length direction of the first work platform 1 by a second crimping motor 74, so as to realize the sequential docking of the crimping assembly to each of the plurality of integrated micro-droplet chips 100. It can be understood that corresponding guide rails are also provided in the crimping assembly to guide the smooth lifting and lateral movement of the detection crimping block 71; in a preferred embodiment, a light source 77 is also provided on the moving block 72 to provide a light field for the camera of the fluorescence detection module 6. The light source 77 is arranged on the moving block 72, so that the relative position of the light source 77 and the detection crimping block 71 is stable, and both are moved synchronously, and the control is simplified.

[0052] In some embodiments, the detection crimping block 71 is formed with a first detection through hole and a second detection through hole corresponding to the chip oil hole 101 and the chip air-liquid hole 102 respectively, the detection oil enters the reaction chamber 103 through the first detection through hole and the second detection through hole, and the detection crimping block 71 has a first sealing gasket 75 for sealing the crimping position when the detection crimping block 71 is crimped with the droplet generation module 3.

[0053] Continuing to refer to Figure 3 and Figure 4 As shown, in some embodiments, the first work platform 1 and the second work platform 2 are each vertically provided with a guide slide rod 121 at the corresponding four corner regions, in a specific embodiment, the guide slide rod 121 is in sliding connection with the first work platform 1 and / or the second work platform 2 through a linear bearing 126, a first return spring 122 is sleeved on the outer periphery of the guide slide rod 121, the first return spring 122 is clamped between the first work platform 1 and the second work platform 2, so as to be able to exert a restoring force on the first work platform 1 and the second work platform 2 to separate them from each other, and further comprising a pressing structure (not labeled in the figure), which can overcome the elastic force of the first return spring 122 under the action of a pressing force to switch the second work platform 2 from the chip picking and placing state to the working state. The aforementioned pressing force can be realized by manual pressing by an operator.

[0054] The technical scheme is characterized in that, by applying force to the pressing structure to overcome the elastic force of the first reset spring 122, the first work platform 1 and the second work platform 2 are brought close to each other, and the liquid droplet generating module 3 and the integrated micro-liquid droplet chip 100 are pressed together; when the pressing structure is reversely forced, that is, an upward pulling force is applied, the first reset spring 122 can help the second work platform 2 switch from the working state to the chip picking and placing state, and prevent the pressing structure from falling down under the action of gravity, thereby facilitating the picking and placing of the chip.

[0055] Specifically referring to Figure 4 As shown in the drawings, in some embodiments, the pressing structure includes a pressing rod 123 and a roller 124 hinged at both ends of the second work platform 2; a traction rope 125 (for example, a steel wire) is connected between the pressing rod 123 and the first work platform 1; the traction rope 125 forms an included angle with the roller 124 (that is, the traction rope 125 partially wraps the outer circumferential surface of the roller 124); when the pressing rod 123 is pressed towards the side of the first work platform 1, the traction rope 125 can pull the first work platform 1 and the second work platform 2 close to each other and into the working state; preferably, when the pressing rod 123 is pressed to the lowest position towards the side of the first work platform 1, the connection point of the traction rope 125 and the second work platform 2 is at the center point of the roller 124 close to the side of the first work platform 1.

[0056] In the technical scheme, the state switching of the second work platform 2 can be driven when the position of the pressing rod 123 changes through the change direction guiding action of the roller 124 and the pulling connection action of the rope, and the structure is simple; in particular, after the pressing rod 123 is pressed, the connection point of the traction rope 125 and the pressing rod 123 is below the center point of the roller 124, thereby realizing self-locking of the structure, and a separate locking device is not required, further simplifying the structure design.

[0057] Specifically referring to Figure 5 As shown in the drawings, the liquid droplet generating module 3 includes a generating pressing block 33, the length extension direction of the generating pressing block 33 is parallel to the width direction of the second work platform 2, and both ends of the generating pressing block 33 are connected to the second work platform 2 via connecting lugs (not labeled in the drawings); the generating first hole 31 and the generating second hole 32 are both formed on the generating pressing block 33; the liquid droplet generating module 3 further includes a generating cover plate 34, the generating cover plate 34 has a top surface that seals the chip oil hole 101 and the chip gas-liquid hole 102 through a cover plate sealing gasket 310 when generating micro-liquid droplets, and realizes the cover closing position of the air path (not labeled in the drawings) on the generating cover plate 34 and the pressing connection communication of the chip gas-liquid hole 102.

[0058] In the technical solution, the two ends of the generating block 33 are connected with the second operation platform 2 via the connecting lifting lugs, and the structure is reliable and simple.

[0059] In some embodiments, the generating cover plate 34 is hinged with the generating block 33 through a rotating shaft 36, and the generating block 33 is provided with an electromagnet 35, and when the electromagnet 35 is powered, the generating cover plate 34 is attracted to rotate around the rotating shaft 36 to be in the closed position. In the technical solution, the electromagnet 35 is used to attract the generating cover plate 34, and the structure and control are relatively simple.

[0060] In some embodiments, the electromagnet 35 has two, and the two electromagnets 35 are respectively located at the two ends of the length of the generating block 33, and the two ends of the length of the generating cover plate 34 are respectively provided with an attracting block corresponding to the position of each electromagnet 35. The two electromagnets 35 respectively located at the two ends of the length of the generating block 33 can ensure the smooth closing of the generating cover plate 34, and can ensure the reliable sealing of the generating cover plate 34.

[0061] In some embodiments, the rotating shaft 36 is sleeved with a second reset spring 37 for switching the generating cover plate 34 from the closed position to the open position when the electromagnet 35 is powered off. In the technical solution, the second reset spring 37 is used to open the generating cover plate 34 by its own elastic force when the electromagnet 35 is powered off. The aforementioned second reset spring 37 can be a torsional spring.

[0062] In some embodiments, a hand-held protrusion (not shown in the figure) is arranged on the side edge of the generating cover plate 34 away from the rotating shaft 36, so as to facilitate the operator to exert force on the hand-held protrusion to assist in opening or closing the generating cover plate 34. A cover plate sealing gasket 310 is arranged on the side end face of the generating cover plate 34 facing the generating block 33, and the cover plate sealing gasket 310 is arranged around each of the generating first hole 31 and the generating second hole 32.

[0063] In some embodiments, the two ends of the length of the generating block 33 are vertically slidably connected with the second operation platform 2 via the connecting lifting lugs, and a guide rod 38 is arranged between the top surface of the generating block 33 and the bottom surface of the second operation platform 2, and a third reset spring 39 is sleeved outside the guide rod 38, and the third reset spring 39 is clamped between the bottom surface of the second operation platform 2 and the top surface of the generating block 33. In the technical solution, the two ends of the generating block 33 are vertically slidably connected with the second operation platform 2 via the connecting lifting lugs, and the third reset spring 39 clamped between the second operation platform 2 and the generating block 33 can buffer and adjust the horizontal state of the generating block 33, thereby ensuring the pressure sealing property of the generating block 33 to the integrated micro-droplet chip 100.

[0064] Specifically,Figure 6 As shown in the drawings, in some embodiments, the PCR amplification module 4 comprises a semiconductor heating element 41, a heat-conducting block 43 is connected to a first end of the semiconductor heating element 41, a temperature control cavity is formed on an end face of the heat-conducting block 43 away from the semiconductor heating element 41, and a heat dissipation structure (not shown in the drawings) is connected to a second end of the semiconductor heating element 41. The heat-conducting block 43 can uniformly transmit the heat of the semiconductor heating element 41 to the temperature control cavity, thereby uniformly controlling the temperature of the microdroplets in the reaction chamber 103. The semiconductor heating element 41 specifically adopts a Peltier element, and the number of Peltier elements can be multiple according to actual needs.

[0065] In some embodiments, the heat dissipation structure comprises a first heat dissipation block 44 connected in contact with the second end and a second heat dissipation block 48 connected in contact with the first heat dissipation block 44, and the heat dissipation area of the second heat dissipation block 48 is greater than that of the first heat dissipation block 44, so as to quickly dissipate the heat of the first end of the semiconductor heating element 41.

[0066] In some embodiments, the heat dissipation structure further comprises a third heat dissipation block 47 and a heat-conducting pipe 49, and the heat-conducting pipe 49 specifically adopts a heat pipe having a heat-conducting medium therein and a high heat transfer efficiency. The first end of the heat-conducting pipe 49 is between the first heat dissipation block 44 and the second heat dissipation block 48, and the second end of the heat-conducting pipe 49 is in heat exchange connection with the third heat dissipation block 47. In this technical solution, the heat at the first heat dissipation block 44 and the second heat dissipation block 48 can be quickly transferred and conducted to the third heat dissipation block 47 for dissipation. The length of the heat-conducting pipe 49 can be adjusted to flexibly select the setting position of the third heat dissipation block 47, thereby greatly increasing the heat dissipation area and achieving the purpose of quickly dissipating heat.

[0067] In some embodiments, the first heat dissipation block 44 is annularly provided with a heat insulation plate 42 at the connection position with the semiconductor heating element 41, so as to prevent the mixing of cold and hot heat and reduce energy consumption; and / or, the heat-conducting block 43 is provided with a temperature probe (not shown in the drawings) and a temperature fuse (not shown in the drawings). The temperature probe can detect the real-time temperature of the heat-conducting block 43, and the temperature fuse can disconnect the power supply of the semiconductor heating element 41 when the real-time temperature of the heat-conducting block 43 exceeds a preset value, thereby achieving the purpose of constant temperature regulation.

[0068] In some embodiments, the chip placing grooves 13 are sequentially and spacedly arranged along the length direction of the first workbench 1, and each of the droplet generating module 3 and the PCR amplification module 4 is arranged in one-to-one correspondence with each of the chip placing grooves 13, thereby improving the processing throughput of the instrument.

[0069] According to the embodiment of the present application, a control method of the compact digital PCR instrument is also provided, comprising the following steps:

[0070] The integrated micro-droplet chip 100 carrying the sample liquid is placed in the chip placing groove 13;

[0071] The second operation platform 2 is controlled to switch from the chip taking and placing state to the working state;

[0072] The droplet generating module 3 is controlled to operate to generate micro-droplets in the integrated micro-droplet chip 100 and store the micro-droplets in the reaction chamber 103 of the integrated micro-droplet chip 100;

[0073] After the micro-droplets are generated, the first operation platform 1 and the second operation platform 2 are controlled to be synchronously flipped by 180°, and then the PCR amplification module 4 is controlled to operate to amplify the micro-droplets in the reaction chamber 103;

[0074] After the amplification is completed, the detection pressing module 7 and the fluorescence detection module 6 are controlled to move towards the integrated micro-droplet chip 100, and after the detection pressing module 7 is docked with the generating first hole 31 and the generating second hole 32 of the droplet generating module 3, the detection pressing module 7 is controlled to operate to drive the amplified micro-droplets to enter the sample cavity 105 of the integrated micro-droplet chip 100 from the reaction chamber 103 through the fluorescence detection area 104 of the integrated micro-droplet chip 100, and the fluorescence detection module 6 is controlled to perform fluorescence detection at the fluorescence detection area 104.

[0075] The technical scheme integrates the droplet generating module 3 and the PCR amplification module 4 on the second operation platform 2, and the second operation platform 2 has the working state and the chip taking and placing state and can switch between the two states, and the synchronous flipping of the first operation platform 1 and the second operation platform 2 is realized by the rotation driving of the rotating mechanism 5, so that the integrated micro-droplet chip 100 does not need to be switched between different operation modules or flipped by using the scheduling mechanism in the prior art, the structure of the digital PCR instrument is more compact, the volume of the instrument is smaller, the portability is stronger, and the manufacturing cost is relatively reduced

[0076] In some embodiments, the control method further comprises: during the operation of the PCR amplification module 4, controlling the first work platform 1 and the second work platform 2 to synchronously reciprocate, which can accelerate the uniformity of the temperature field by fluid convection, improve the heat transfer efficiency and temperature uniformity in the micro-droplet sample, and reduce the temperature uniformity requirement of the temperature control module itself, thereby improving the efficiency of the amplification link; and / or after the fluorescence detection is completed, controlling the first work platform 1 and the second work platform 2 to synchronously rotate 180° in reverse, and then controlling the second work platform 2 to switch from the working state to the chip taking and placing state.

[0077] It is easy for those skilled in the art to understand that the above advantageous modes can be freely combined and superimposed without conflict.

[0078] The above is only the preferred embodiment of the present application, and should not be used to limit the present application. Any modification, equivalent replacement and improvement made within the spirit and principle of the present application shall be included in the protection scope of the present application. The above is only the preferred embodiment of the present application, and should not be used to limit the present application. Any modification, equivalent replacement and improvement made within the spirit and principle of the present application shall be included in the protection scope of the present application.

Claims

1. A compact digital PCR instrument, characterized by, The integrated module comprises a first work platform (1) and a second work platform (2), the first work platform (1) has a plurality of chip placement grooves (13) for positioning and placing an integrated micro-droplet chip (100), the second work platform (2) is arranged in parallel with the first work platform (1) and has a working state close to the first work platform (1) and a chip pick-and-place state away from the first work platform (1), the second work platform (2) has a droplet generation module (3) and a PCR amplification module (4), when the second work platform (2) is in the working state, the droplet generation module (3) and chip oil holes (101) and chip gas-liquid holes (102) of the integrated micro-droplet chip (100) are respectively pressed and connected to form micro-droplets in the integrated micro-droplet chip (100), and reaction chambers (103) of the integrated micro-droplet chip (100) are in a temperature control cavity of the PCR amplification module (4), and the rotating mechanism (5) is used for driving the first work platform (1) and the second work platform (2) to synchronously turn over 180° when the second work platform (2) is in the working state, so that the reaction chambers (103) are upside down to ensure that the micro-droplets stored in the reaction chambers (103) can be at the bottom of the reaction chambers (103) under the action of gravity. The integrated module further comprises a fluorescence detection module (6) on the first side and a detection compression module (7) on the second side, the first side and the second side are one of the top side and the bottom side of the first work platform (1), the fluorescence detection module (6) can be controlled to rise and fall to realize fluorescence detection corresponding to the position of the fluorescence detection area (104) of the integrated micro-droplet chip (100) by moving away from or close to the first work platform (1), the detection compression module (7) can be controlled to rise and fall to realize compression connection with the generation first hole (31) and the generation second hole (32) in the droplet generation module (3) by moving close to or away from the second work platform (2), so that the detection oil in the detection oil way (70) can enter the reaction chambers (103) to make the micro-droplets after amplification reaction float to the fluorescence detection area (104) and enter the sample cavity (105) of the integrated micro-droplet chip (100).

2. The compact digital PCR instrument of claim 1, wherein, The integrated module further comprises left and right spaced vertical plates (11), the length ends of the first work platform (1) are pivotally arranged on the top ends of the two vertical plates (11), the bottom ends of the two vertical plates (11) are connected by a connecting plate (12), the rotating mechanism (5) comprises a rotary motor (51) and a transmission assembly (52), the rotary motor (51) is fixedly connected to one of the vertical plates (11), and the rotary motor (51) is drivingly connected to one end of the first work platform (1) through the transmission assembly (52) to drive the first work platform (1) to rotate by a preset angle.

3. The compact digital PCR instrument of claim 2, wherein, Further comprising a device bottom plate (201) and a longitudinal moving assembly, the longitudinal moving assembly comprises a longitudinal moving guide rail (2021) and a longitudinal moving motor (2022), the bottom end of the integrated module is slidingly connected between the longitudinal moving guide rail (2021) and the device bottom plate (201), and the longitudinal moving motor (2022) can drive the integrated module to reciprocatingly move linearly along the guide direction of the longitudinal moving guide rail (2021), and the guide direction of the longitudinal moving guide rail (2021) is a horizontal direction perpendicular to the length direction of the first work platform (1).

4. The compact digital PCR instrument of claim 3, wherein, The detection pressure connection module (7) comprises an air cavity (76) and a pressure connection assembly connected to the air cavity (76), the air cavity (76) is used for providing negative pressure for the liquid drop generation module (3) and providing positive pressure for the detection pressure connection module (7); and / or the fluorescence detection module (6) and the detection pressure connection module (7) can be controlled to move to correspond to the positions of the integral micro liquid drop chips (100).

5. The compact digital PCR instrument of claim 4, wherein, The pressure connection assembly comprises a detection pressure connection block (71) and a moving block (72), the detection pressure connection block (71) and the moving block (72) are slidingly connected and can be driven to lift by a first pressure connection motor (73), and the moving block (72) is slidingly connected with the air cavity (76) and can be driven to move horizontally along the length direction of the first work platform (1) by a second pressure connection motor (74).

6. The compact digital PCR instrument of claim 5, wherein, The moving block (72) is provided with a light source (77) to provide a light field for a camera of the fluorescence detection module (6).

7. The compact digital PCR instrument of claim 1, wherein, The four corner regions of the first work platform (1) and the second work platform (2) corresponding to each other are vertically provided with guide sliding rods (121), the outer periphery of the guide sliding rod (121) is sleeved with a first return spring (122), the first return spring (122) is clamped between the first work platform (1) and the second work platform (2), and further comprising a pressing structure, the pressing structure can overcome the elastic force of the first return spring (122) under the action of a pressing force to make the second work platform (2) switch from the chip taking and placing state to the working state.

8. The compact digital PCR instrument of claim 3, wherein, The droplet generating module (3) comprises a generating block (33), the length direction of the generating block (33) is parallel to the width direction of the second operation platform (2), and the length ends of the generating block (33) are connected with the second operation platform (2) through connecting lugs, the generating first hole (31) and the generating second hole (32) are formed on the generating block (33), and the droplet generating module (3) further comprises a generating cover plate (34), the generating cover plate (34) has a cover position for sealing the top surface of the chip oil hole (101) and the chip gas-liquid hole (102) when generating microdroplets and realizing the pressure connection communication between the air path on the generating cover plate (34) and the chip gas-liquid hole (102).

9. A method of controlling a compact digital PCR machine according to any one of claims 1 to 8, characterized by, The method comprises the following steps: placing an integrated microdroplet chip (100) loaded with a sample liquid into the chip placing groove (13); controlling the second operation platform (2) to switch from a chip taking and placing state to a working state; controlling the droplet generating module (3) to operate to generate microdroplets in the integrated microdroplet chip (100) and store the microdroplets in the reaction chamber (103) of the integrated microdroplet chip (100); after the microdroplet generation is completed, controlling the first operation platform (1) and the second operation platform (2) to synchronously overturn by 180°, and controlling the PCR amplification module (4) to operate to amplify the microdroplets in the reaction chamber (103); after the amplification is completed, controlling the detection pressure connection module (7) and the fluorescence detection module (6) to move towards the integrated microdroplet chip (100), after the detection pressure connection module (7) is butt-jointed with the generating first hole (31) and the generating second hole (32) of the droplet generating module (3), controlling the detection pressure connection module (7) to operate to drive the amplified microdroplets to enter the sample adding cavity (105) of the integrated microdroplet chip (100) from the reaction chamber (103) through the fluorescence detection area (104) of the integrated microdroplet chip (100), and controlling the fluorescence detection module (6) to perform fluorescence detection at the fluorescence detection area (104).

10. The control method according to claim 9, characterized by, Further comprising: controlling the first operation platform (1) and the second operation platform (2) to synchronously reciprocate during the operation of the PCR amplification module (4); and / or, after the fluorescence detection is completed, controlling the first operation platform (1) and the second operation platform (2) to synchronously rotate by 180° in the reverse direction, and then controlling the second operation platform (2) to switch from the working state to the chip taking and placing state.

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