Integrally-controlled gene amplification equipment

Abstract

The invention relates to the field of biological devices, in particular to integrally-controlled gene amplification equipment which comprises a shell. A placement cavity is formed in the shell; a placement device is arranged in the placement cavity; the placing device comprises a placing shell, a separating assembly, a centrifugal frame and a circulating assembly; the separation assembly is used for separating the amplification reaction cavity into three fan-shaped reaction chambers; the three reaction chambers are all provided with constant-temperature assemblies; the three constant-temperature reaction chambers are arranged, a sample needing to be amplified is driven by the circulating assembly to rotate in the amplification reaction cavity around the axis of the containing shell at a constant speed, the sample is seamlessly switched between the two adjacent reaction chambers with different preset temperatures, and compared with a traditional mode that the temperature is increased and decreased through a cooling module to achieve different reaction temperature directions, the sample amplification efficiency is improved. The equipment does not need to be provided with a cooling module, so that the temperature rising and falling matching time in the sample amplification reaction process is saved, the reaction temperature control is more accurate, and the gene sample amplification efficiency is improved.

Description

Technical Field

[0001] The invention relates to the field of biological devices, and in particular to an integrated controlled gene amplification device. Background Art

[0002] Gene amplification is a biological process that refers to the process of increasing the number of copies of a specific gene in a cell. The commonly used gene amplification equipment in the laboratory is the polymerase chain reaction (PCR) instrument. Through this gene amplification equipment, researchers can specifically amplify trace amounts of DNA fragments and other gene samples. The PCR instrument is a key device for polymerase chain reaction (PCR). It achieves denaturation, annealing (renaturation) and extension of gene samples by controlling temperature changes and the specific time that the gene sample stays at a specific temperature. The gene amplification device takes about two hours to perform an amplification (thirty cycles). The working process of the gene amplification instrument is approximately that the sample stays at 94°C for 5 minutes for pre-denaturation, and then a single cycle is started. Among them, the temperature and time required for each program in a single cycle are usually: denaturation, stay at 94°C for 30 seconds; renaturation, stay at 55°C for 30 seconds; extension, stay at 72°C for 1 minute.

[0003] Existing gene amplification instruments are equipped with temperature regulating devices such as heating devices and cooling devices to adjust the temperature of the environment around the sample, thereby achieving denaturation, annealing and extension of the gene sample. However, the temperature in the gene amplification instrument takes a certain amount of time to rise and fall, which affects the efficiency of gene amplification. Summary of the invention

[0004] The present invention provides an integrated controlled gene amplification device to solve the above problems.

[0005] The present invention provides an integrated controlled gene amplification device, which adopts the following technical scheme: an integrated controlled gene amplification device, comprising a housing; a placement cavity is provided in the housing; a placement device is provided in the placement cavity; The placement device includes a placement shell, a partition assembly, a centrifugal frame, and a circulation assembly; The placement shell is in the shape of a cylinder with its axis vertically arranged, and a cylindrical amplification reaction chamber is arranged inside; The partition component is used to divide the amplification reaction chamber into three fan-shaped reaction chambers; the three reaction chambers are concentrically arranged; the three reaction chambers are sequentially arranged as a denaturation chamber, a renaturation chamber and an extension chamber along the circumference of the placement shell; the denaturation chamber, the renaturation chamber and the extension chamber are all provided with constant temperature components to maintain the temperature of the corresponding reaction chamber at the required preset temperature, for example, the temperature in the denaturation chamber is maintained at 94°C; the temperature in the renaturation chamber is maintained at 55°C; the temperature in the extension chamber is maintained at 72°C; the denaturation chamber is provided with a sample loading port, and the renaturation chamber is provided with a sample outlet; the ratio of the radian values ​​of the three reaction chambers to the required residence time of the sample in the corresponding reaction chamber is the same; that is, the radian value of the denaturation chamber is set to m, the radian value of the renaturation chamber is set to n and the radian value of the extension chamber is set to p, and the required residence time of the sample in the three reaction chambers is set to x, y and z, respectively, then m / x=n / y=p / z; There are multiple centrifugal racks, which are arranged in the amplification reaction chamber; the centrifugal racks include a placement plate; the placement plate is arranged in a direction perpendicular to the radial direction of the placement shell; the placement plate is provided with multiple placement holes for placing centrifuge tubes containing samples; The circulation component is arranged in the amplification reaction chamber, and is used to drive the centrifugal frame to drive the sample to rotate at a constant speed around the axis of the placement shell in the amplification reaction chamber, and to make the sample pass through the denaturation chamber, renaturation chamber, and extension chamber in sequence. By setting up three constant temperature reaction chambers, the sample to be amplified is driven by the circulation component to rotate at a constant speed around the axis of the placement shell in the amplification reaction chamber, and the sample seamlessly switches between two adjacent reaction chambers with different preset temperatures. Compared with the traditional method of raising and lowering the temperature through the cooling module to achieve different reaction temperatures, this device does not need to set up a cooling module, which saves the time of temperature rise and fall matching during the sample amplification reaction, makes the temperature control of the reaction more accurate, and improves the efficiency of gene sample amplification.

[0006] Furthermore, the partition assembly includes three partitions; the partitions are vertically arranged and radially extended along the placement shell; the upper end of the partitions is fixed to the upper cavity wall of the amplification reaction chamber, and the lower end is abutted against the lower cavity wall of the amplification reaction chamber; the end of the partition away from the axis of the placement shell is abutted against the side cavity wall of the amplification reaction chamber, and the end close to the axis of the placement shell is fixedly connected with a connecting ring; the connecting ring and the placement shell are coaxially arranged; the upper end of the connecting ring is fixed to the upper cavity wall of the amplification reaction chamber, and the lower end is abutted against the lower cavity wall of the amplification reaction chamber; the partition divides the amplification reaction chamber into a denaturation chamber, a renaturation chamber and an extension chamber.

[0007] Furthermore, the arc value of the denaturation chamber is 90°, the arc value of the renaturation chamber is 90°, and the arc value of the extension chamber is 180°. The sample residence time in the three reaction chambers is set to be 30 seconds in the denaturation chamber, 30 seconds in the renaturation chamber, and 1 minute in the extension chamber.

[0008] Furthermore, the centrifugal frame also includes a push plate; two push plates are provided, distributed at both ends of the placement plate along the tangent direction perpendicular to the radial direction of the placement shell; the push plate is fixed to the end of the placement plate; a clamping protrusion is fixed at one end of the push plate away from the placement plate.

[0009] Further, the circulation assembly includes a mounting groove, a rotating disk, a retaining ring, and a guide structure; The mounting groove is an annular groove; the mounting groove is a through groove arranged on the side wall of the connecting ring; the mounting groove and the connecting ring are coaxial; The turntable and the connecting ring are coaxially arranged; the turntable is rotatably installed in the installation groove; the turntable is connected to a servo motor; the servo motor drives the turntable to rotate at a uniform speed; a plurality of card blocks are distributed in an annular manner on the side wall of the turntable; the card blocks are spaced at a preset interval; the preset interval is equal to the length of the placement plate; a card slot is provided on the side wall of the card block away from the axis of the turntable; the card slot is trapezoidal in shape with the small end facing the axis of the turntable; The retaining ring is coaxially arranged outside the turntable; a circular circulation gap is formed between the retaining ring and the turntable; the retaining ring is fixed on the partition plate; a sample loading gap and a sample discharging gap are arranged on the retaining ring; the sample loading gap corresponds to the sample loading port, and the sample discharging gap corresponds to the sample discharging port; The guiding structure is used to place the centrifugal frame in the amplification reaction chamber, so that the push plates at both ends of the placement plate are stuck between two adjacent blocks, and the corresponding clamping protrusions are stuck in the clamping grooves, so that the turntable drives the centrifugal frame to rotate around the turntable axis in the circulation gap through the clamping blocks. After the amplification reaction is completed, the centrifugal frame is driven away from the clamping blocks, the push plates and the clamping blocks are separated, and the clamping protrusions are separated from the clamping grooves.

[0010] Further, the guiding structure includes a sample loading guiding rod and a sample discharging guiding rod; Two sample loading guide rods are provided, which are horizontally distributed on both sides of the sample loading gap and are arranged in the sample loading port; the sample loading guide rods are arranged along the radial direction of the turntable, and are initially arranged horizontally; one end of the sample loading guide rod close to the axis of the turntable is rotatably mounted on the outer wall of the retaining ring through a horizontally arranged rotating shaft; a sample loading slide groove is provided on the side wall of the sample loading guide rod close to the sample loading gap; the clamping protrusion and the sample loading slide groove are slidably matched; the push plate and the side wall of the sample loading guide rod close to the sample loading gap are abutted and slidably matched; Two sample-discharging guide rods are provided, which are horizontally distributed on both sides of the sample-discharging gap and are arranged in the sample-discharging port; the sample-discharging guide rods are arranged along the radial direction of the turntable, and initially, the end of the sample-discharging guide rod away from the axis of the turntable is higher than the end close to the axis of the turntable; the end of the sample-discharging guide rod close to the axis of the turntable is rotatably mounted on the outer side wall of the retaining ring through a horizontally arranged rotating shaft; a sample-discharging groove is provided on the side wall of the sample-discharging guide rod close to the sample-loading gap; the clamping protrusion and the sample-discharging groove are slidably matched; the push plate and the side wall of the sample-discharging guide rod close to the sample-loading gap are abutted and slidably matched.

[0011] Further, a loading motor and a discharging motor are fixed on the retaining ring; the output shaft of the loading motor is fixedly connected to the rotating shaft of the loading guide rod; the output shaft of the discharging motor is fixedly connected to the rotating shaft of the discharging guide rod. When the amplification sample needs to be added after the amplification starts, the card convex of the centrifugal rack of the sample to be added is first slidably installed in the loading slide groove, the loading motor drives the loading guide rod to keep horizontal, so that the centrifugal rack of the sample to be added is pre-denatured in the denaturation chamber, after a period of pre-denaturation, the loading motor drives the loading guide rod to rotate at one end away from the axis of the turntable, so that the centrifugal rack of the sample to be added slides along the loading guide rod to the circulation gap under the driving of its own gravity, and when the placement plate of the centrifugal rack corresponds to the gap between two adjacent card blocks, the centrifugal rack slides between the two card blocks, and the card convex slides into the card slot, the push plate and the card block abut, when the card block rotates with the turntable, the card block drives the centrifugal rack to drive the sample to rotate around the turntable axis in the circulation gap, and passes through the denaturation chamber, the renaturation chamber, and the extension chamber in sequence for amplification reaction. When the sample completes a predetermined number of amplification cycles, the sample outlet motor drives the sample outlet guide rod to rotate, so that the end of the sample outlet guide rod away from the turntable axis is lower than the end close to the turntable axis, so that the centrifugal frame carrying the sample that has completed amplification is away from the block, the push plate and the block are disengaged, and the card protrusion is disengaged from the card slot, so that the centrifugal frame carrying the sample that has completed amplification leaves the amplification reaction chamber from the sample outlet.

[0012] Furthermore, a proximity switch is provided between adjacent card blocks; a controller is provided in the housing; and the proximity switch and the controller are electrically connected. When the centrifugal rack enters between two card blocks, the proximity switch is triggered, providing a signal to the controller and starting counting. When the centrifugal rack rotates to complete the preset number of amplification cycles, the controller controls the sample output motor to drive the sample output guide rod to rotate, so that the centrifugal rack carrying the amplified sample leaves the amplification reaction chamber from the sample output port, so that new samples to be amplified can be added or samples that have been amplified can be taken out at any time, thereby improving the overall amplification efficiency.

[0013] Furthermore, a sample loading cover is installed on the sample loading port; and a sample outlet cover is installed on the sample outlet to ensure sealing and reduce heat loss in the amplification reaction chamber.

[0014] Furthermore, an upper cover is provided at the upper end of the placement cavity; and an operation screen is provided at one side of the shell.

[0015] The beneficial effects of the present invention are: 1. By setting three constant temperature reaction chambers, the sample to be amplified rotates at a uniform speed around the axis of the placement shell in the amplification reaction chamber driven by the circulation component, and the sample seamlessly switches between two adjacent reaction chambers with different preset temperatures. The ratio of the curvature values ​​of the three reaction chambers to the required retention time of the sample in the corresponding reaction chamber is the same. Compared with the existing method of controlling the retention time of the sample in the corresponding reaction chamber by a controller, the retention time of the sample in the corresponding reaction chamber is controlled by the curvature of the reaction chamber, which is convenient for amplifying multiple samples at the same time and improves the amplification efficiency. Compared with the traditional method of raising and lowering the temperature through a cooling module to achieve different reaction temperatures, the device does not need to set a cooling module, which saves the time for temperature rising and falling matching during the sample amplification reaction, makes the temperature control of the reaction more accurate, and improves the efficiency of gene sample amplification.

[0016] 2. When the centrifuge frame enters between the two blocks, the proximity switch is triggered to provide a signal to the controller and start counting. When the centrifuge frame rotates to complete the preset number of amplification cycles, the controller controls the sample outlet motor to drive the sample outlet guide rod to rotate, so that the centrifuge frame carrying the amplified sample leaves the amplification reaction chamber from the sample outlet, so that new samples to be amplified can be added or samples that have been amplified can be taken out at any time, thereby improving the overall amplification efficiency. BRIEF DESCRIPTION OF THE DRAWINGS

[0017] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required for use in the embodiments or the description of the prior art will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative labor.

[0018] Figure 1 It is a structural schematic diagram of an embodiment of an integrated controlled gene amplification device of the present invention; Figure 2 A top view of an embodiment of an integrated controlled gene amplification device of the present invention with the upper cover removed; Figure 3 An exploded view of a placement device of an embodiment of an integrated controlled gene amplification device of the present invention; Figure 4 A schematic diagram of a placement shell of an embodiment of an integrated controlled gene amplification device of the present invention; Figure 5 It is a schematic diagram of a separation component, a retaining ring, and a guide structure of an embodiment of an integrated controlled gene amplification device of the present invention; Figure 6 A schematic diagram of a turntable and a centrifugal rack of an embodiment of an integrated controlled gene amplification device of the present invention; Figure 7 A schematic diagram of a turntable of an embodiment of an integrated controlled gene amplification device of the present invention; Figure 8 A top view of a placement device of an embodiment of an integrated controlled gene amplification device of the present invention; Fig. 9 for Figure 8 Sectional view at AA in the middle; Fig.10 A side view of a placement device of an embodiment of an integrated controlled gene amplification device of the present invention; Fig.11 for Fig.10 Sectional view at the middle BB; Fig.12 A schematic diagram of a centrifugal rack of an embodiment of an integrated controlled gene amplification device of the present invention; Fig.13 A cross-sectional view of another angle of a placement device of an embodiment of an integrated controlled gene amplification device of the present invention; Fig.14 for Fig.13 Enlarged view of point C in the middle.

[0019] In the figure: 100, centrifuge tube; 200, shell; 210, operation screen; 220, upper cover; 300, placement shell; 310, denaturation chamber; 311, sample loading port; 320, renaturation chamber; 321, sample outlet; 330, extension chamber; 410, partition; 420, connecting ring; 500, centrifuge rack; 510, placement plate; 520, push plate; 530, clamping cam; 600, turntable; 610, clamping block; 611, clamping slot; 612, proximity switch; 700, retaining ring; 710, circulation gap; 720, sample loading gap; 730, sample outlet gap; 740, sample loading guide rod; 741, sample loading slide; 750, sample outlet guide rod; 751, sample outlet slide. DETAILED DESCRIPTION

[0020] In the description of the present invention, it should be noted that, unless otherwise clearly specified and limited, the terms "installed", "connected", and "connected" should be understood in a broad sense, for example, it can be a fixed connection, a detachable connection, or an integral connection; it can be a mechanical connection or an electrical connection; it can be a direct connection, or it can be indirectly connected through an intermediate medium, or it can be the internal communication of two components. For ordinary technicians in this field, the specific meanings of the above terms in the present invention can be understood according to specific circumstances.

[0021] An embodiment of an integrated controlled gene amplification device of the present invention is as follows Figures 1 to 14As shown: an integrated controlled gene amplification device, including a housing 200; a placement cavity is provided in the housing 200; a placement device is provided in the placement cavity; an upper cover 220 is provided at the upper end of the placement cavity; an operation screen 210 is provided on one side of the housing 200. The placement device includes a placement shell 300, a partition assembly, a centrifugal frame 500, and a circulation assembly; The placement shell 300 is in the shape of a cylinder with a vertical axis, and a cylindrical amplification reaction chamber is arranged inside the placement shell 300; the placement shell 300 is fixed on the outer shell 200.

[0022] The partition assembly is used to divide the amplification reaction chamber into three fan-shaped reaction chambers; the three reaction chambers are arranged concentrically; the three reaction chambers are arranged in sequence along the circumference of the placement shell 300 as a denaturation chamber 310, a renaturation chamber 320 and an extension chamber 330; the denaturation chamber 310, the renaturation chamber 320 and the extension chamber 330 are all provided with a constant temperature assembly to maintain the temperature of the corresponding reaction chamber at the required preset temperature, for example, the temperature in the denaturation chamber 310 is maintained at 94°C; the temperature in the renaturation chamber 320 is maintained at 55°C; the temperature in the extension chamber 330 is maintained at 72°C. The constant temperature assembly is an existing technology on the PCR instrument, so it will not be repeated. The denaturation chamber 310 is provided with a sample loading port 311, and the renaturation chamber 320 is provided with a sample outlet 321; a sample loading cover is installed on the sample loading port 311; and a sample outlet cover is installed on the sample outlet 321 to ensure sealing and reduce heat loss in the amplification reaction chamber.

[0023] The ratio of the radian values ​​of the three reaction chambers to the time the sample needs to stay in the corresponding reaction chamber is the same; that is, the radian value of the denaturation chamber 310 is set to m, the radian value of the renaturation chamber 320 is set to n, and the radian value of the extension chamber 330 is set to p, and the sample needs to stay in the three reaction chambers for x in the denaturation chamber 310, y in the renaturation chamber 320, and z in the extension chamber 330, then m / x=n / y=p / z; the radian value of the denaturation chamber 310 is set to 90°, the radian value of the renaturation chamber 320 is set to 90°, and the radian value of the extension chamber 330 is set to 180°, and the sample needs to stay in the three reaction chambers for 30 seconds in the denaturation chamber 310, 30 seconds in the renaturation chamber 320, and 1 minute.

[0024] The partition assembly includes three partitions 410; the partitions 410 are vertically arranged and radially extended along the placement shell 300; the upper end of the partition 410 is fixed to the upper cavity wall of the amplification reaction chamber, and the lower end is abutted against the lower cavity wall of the amplification reaction chamber; the end of the partition 410 away from the axis of the placement shell 300 is abutted against the side cavity wall of the amplification reaction chamber, and the end close to the axis of the placement shell 300 is fixedly connected with a connecting ring 420; the connecting ring 420 and the placement shell 300 are coaxially arranged; the upper end of the connecting ring 420 is fixed to the upper cavity wall of the amplification reaction chamber, and the lower end is abutted against the lower cavity wall of the amplification reaction chamber; the partition 410 divides the amplification reaction chamber into a denaturation chamber 310, a renaturation chamber 320 and an extension chamber 330.

[0025] There are multiple centrifuge racks 500, which are arranged in the amplification reaction chamber; the centrifuge rack 500 includes a placement plate 510 and a push plate 520; the placement plate 510 is arranged in a direction perpendicular to the radial direction of the placement shell 300; there are multiple placement holes on the placement plate 510 for placing the centrifuge tube 100 containing the sample; there are two push plates 520, which are distributed at both ends of the placement plate 510 along the tangential direction perpendicular to the radial direction of the placement shell 300; the push plate 520 is fixed to the end of the placement plate 510; a clamping protrusion 530 is fixed at the end of the push plate 520 away from the placement plate 510.

[0026] The circulation component is arranged in the amplification reaction chamber, and is used to drive the centrifugal frame 500 to drive the sample to rotate at a uniform speed around the axis of the placement shell 300 in the amplification reaction chamber, and to make the sample pass through the denaturation chamber 310, the renaturation chamber 320, and the extension chamber 330 in sequence. By setting up three constant temperature reaction chambers, the sample to be amplified is driven by the circulation component to rotate at a uniform speed around the axis of the placement shell 300 in the amplification reaction chamber, and the sample seamlessly switches between two adjacent reaction chambers with different preset temperatures. Compared with the traditional method of raising and lowering the temperature through the cooling module to achieve different reaction temperatures, the device does not need to set up a cooling module, which saves the time of temperature rise and fall matching during the sample amplification reaction, makes the temperature control of the reaction more accurate, and improves the efficiency of gene sample amplification.

[0027] The circulation component includes a mounting groove, a turntable 600, a retaining ring 700, and a guide structure; the mounting groove is an annular groove; the mounting groove is a through groove arranged on the side wall of the connecting ring 420; the mounting groove and the connecting ring 420 are coaxial; the turntable 600 and the connecting ring 420 are coaxially arranged; the turntable 600 is rotatably installed in the mounting groove; and a servo motor is connected to the turntable 600. The servo motor drives the turntable 600 to rotate at a uniform speed; a plurality of blocks 610 are distributed in an annular manner on the side wall of the turntable 600; the blocks 610 are spaced at preset intervals; the preset intervals are equal in length to the placement plate 510; a slot 611 is provided on the side wall of the block 610 away from the axis of the turntable 600; the slot 611 is trapezoidal in shape with the small end facing the axis of the turntable 600; the retaining ring 700 is coaxially arranged on the outside of the turntable 600; a ring-shaped circulation gap 710 is formed between the retaining ring 700 and the turntable 600; the retaining ring 700 is fixed on the partition 410; a sample loading gap 720 and a sample discharging gap 730 are provided on the retaining ring 700; the sample loading gap 720 corresponds to the sample loading port 311, and the sample discharging gap 730 corresponds to the sample discharging port 321.

[0028] The guide structure is used to make the push plates 520 at both ends of the placement plate 510 stuck between two adjacent blocks 610 and the corresponding locking protrusions 530 stuck in the locking grooves 611 when the centrifugal rack 500 needs to be placed in the amplification reaction chamber, so that the turntable 600 drives the centrifugal rack 500 to rotate around the axis of the turntable 600 in the circulation gap 710 through the locking blocks 610. After the amplification reaction is completed, the centrifugal rack 500 is driven away from the locking blocks 610, so that the push plates 520 and the locking blocks 610 are separated, and the locking protrusions 530 are separated from the locking grooves 611. The guide structure includes a sample loading guide rod 740 and a sample discharging guide rod 750.

[0029] Two sample loading guide rods 740 are provided, which are horizontally distributed on both sides of the sample loading gap 720 and are arranged in the sample loading port 311; the sample loading guide rods 740 are arranged along the radial direction of the turntable 600, and the sample loading guide rods 740 are initially arranged horizontally; one end of the sample loading guide rod 740 close to the axis of the turntable 600 is rotatably mounted on the outer side wall of the retaining ring 700 through a horizontally arranged rotating shaft; a sample loading chute 741 is provided on the side wall of the sample loading guide rod 740 close to the sample loading gap 720; the clamping protrusion 530 and the sample loading chute 741 are slidably matched; the push plate 520 and the sample loading guide rod 740 are abutted against the side wall of the sample loading gap 720, and are slidably matched; Two sample guide rods 750 are provided, which are horizontally distributed on both sides of the sample gap 730 and are arranged in the sample outlet 321; the sample guide rods 750 are arranged radially along the turntable 600, and initially, the end of the sample guide rod 750 away from the axis of the turntable 600 is higher than the end close to the axis of the turntable 600; the end of the sample guide rod 750 close to the axis of the turntable 600 is rotatably mounted on the outer wall of the retaining ring 700 through a horizontally arranged rotating shaft; a sample chute 751 is provided on the side wall of the sample guide rod 750 close to the sample gap 720; the clamping protrusion 530 and the sample chute 751 are slidably matched; the push plate 520 and the side wall of the sample guide rod 750 close to the sample gap 720 are abutted and slidably matched. The retaining ring 700 is fixed with a sample loading motor and a sample discharging motor; the output shaft of the sample loading motor is fixedly connected to the rotating shaft of the sample loading guide rod 740; the output shaft of the sample discharging motor is fixedly connected to the rotating shaft of the sample discharging guide rod 750.

[0030] When the amplification sample needs to be added after the amplification starts, the protrusion 530 of the centrifugal rack 500 of the sample to be added is first slidably installed in the sample loading slide groove 741, and the sample loading motor drives the sample loading guide rod 740 to remain horizontal, so that the centrifugal rack 500 of the sample to be added is pre-denatured in the denaturation chamber 310. After a period of pre-denaturation, the sample loading motor drives the sample loading guide rod 740 to rotate upward at one end away from the axis of the turntable 600, so that the centrifugal rack 500 of the sample to be added is driven by its own gravity to move along the sample loading guide rod 740 to the upper side of the turntable 600. The circulation gap 710 slides, and when the placement plate 510 of the centrifugal frame 500 corresponds to the gap between two adjacent blocks 610, the centrifugal frame 500 slides between the two blocks 610, and the locking protrusion 530 slides into the locking groove 611, the push plate 520 and the block 610 abut, when the block 610 rotates with the turntable 600, the block 610 drives the centrifugal frame 500 to drive the sample to rotate around the axis of the turntable 600 in the circulation gap 710, and passes through the denaturation chamber 310, the renaturation chamber 320, and the extension chamber 330 in sequence for amplification reaction. When the sample completes a predetermined number of amplification cycles, the sample outlet motor drives the sample outlet guide rod 750 to rotate, so that the end of the sample outlet guide rod 750 away from the axis of the turntable 600 is lower than the end close to the axis of the turntable 600, so that the centrifugal frame 500 carrying the sample that has completed amplification is away from the block 610, the push plate 520 and the block 610 are disengaged, and the latch protrusion 530 is disengaged from the slot 611, so that the centrifugal frame 500 carrying the sample that has completed amplification leaves the amplification reaction chamber from the sample outlet 321.

[0031] A proximity switch 612 is provided between adjacent blocks 610; a controller is provided in the housing 200; and the proximity switch 612 and the controller are electrically connected. When the centrifugal rack 500 enters between two blocks 610, the proximity switch 612 is triggered, providing a signal to the controller and starting counting. When the centrifugal rack 500 rotates to complete the preset number of amplification cycles, the controller controls the sample output motor to drive the sample output guide rod 750 to rotate, so that the centrifugal rack 500 carrying the amplified sample leaves the amplification reaction chamber from the sample outlet 321, so that a new sample to be amplified can be added at any time or a sample that has been amplified can be taken out, thereby improving the overall amplification efficiency.

[0032] In combination with the above embodiments, the use principle and working process of the present invention are as follows: when in use, use a micropipette to add each component in the centrifuge tube 100 in sequence according to the PCR reaction system formula or the instructions of the PCR kit. After all components are added, cover the lid of the centrifuge tube 100. Put the centrifuge tube 100 into the centrifuge and centrifuge for about 10 seconds to concentrate the reaction solution at the bottom of the centrifuge tube 100. Place the centrifuge tube 100 with the configured sample into the placement hole of the placement plate 510. Open the upper cover 220, slide the protrusion 530 of the centrifuge frame 500 into the sample loading chute 741, and the sample loading motor drives the sample loading guide rod 740 to remain horizontal, so that the centrifuge frame 500 of the sample to be added is pre-denatured in the denaturation chamber 310. After a period of pre-denaturation, set the pre-treatment for 5 minutes.

[0033] The loading motor drives the loading guide rod 740 to rotate upward at one end away from the axis of the turntable 600, so that the centrifugal frame 500 with the sample to be added slides toward the circulation gap 710 along the loading guide rod 740 under the drive of its own gravity, and when the placement plate 510 of the centrifugal frame 500 corresponds to the gap between two adjacent blocks 610, the centrifugal frame 500 slides between the two blocks 610, and the protrusion 530 slides into the groove 611, the push plate 520 and the block 610 abut, and when the block 610 rotates with the turntable 600, the block 610 drives the centrifugal frame 500 to drive the sample to rotate around the axis of the turntable 600 in the circulation gap 710, and passes through the denaturation chamber 310, the renaturation chamber 320, and the extension chamber 330 in sequence for amplification reaction. The sample stays in the denaturation chamber 310 for 30 seconds, in the renaturation chamber 320 for 30 seconds, and in the extension chamber 330 for 1 minute.

[0034] When the centrifugal rack 500 enters between the two blocks 610, the proximity switch 612 is triggered, providing a signal to the controller and starting counting. By setting up three constant temperature reaction chambers, the sample to be amplified rotates around the axis of the placement shell 300 at a constant speed in the amplification reaction chamber under the drive of the circulation component, and the sample seamlessly switches between two adjacent reaction chambers with different preset temperatures. Compared with the traditional method of raising and lowering the temperature through the cooling module to achieve different reaction temperatures, the device does not need to set up a cooling module, which saves the time of temperature rise and fall matching during the sample amplification reaction, makes the temperature control of the reaction more accurate, and improves the efficiency of gene sample amplification.

[0035] When the centrifugal rack 500 rotates to complete the preset number of amplification cycles, the controller controls the sample outlet motor to drive the sample outlet guide rod 750 to rotate, so that the end of the sample outlet guide rod 750 away from the axis of the turntable 600 is lower than the end close to the axis of the turntable 600, so that the centrifugal rack 500 carrying the sample that has completed amplification is away from the block 610, the push plate 520 and the block 610 are disengaged, and the latch 530 is disengaged from the slot 611, so that the centrifugal rack 500 carrying the sample that has completed amplification leaves the amplification reaction chamber from the sample outlet 321 and moves to the color development area for subsequent analysis, so as to realize the addition of new samples to be amplified or the removal of samples that have completed amplification at any time, thereby improving the overall amplification efficiency.

[0036] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principle of the present invention should be included in the protection scope of the present invention.

Claims

1. An integrated controlled gene amplification device, characterized in that: It comprises a shell; a placement cavity is provided in the shell; a placement device is provided in the placement cavity; The placement device includes a placement shell, a partition assembly, a centrifugal frame, and a circulation assembly; The placement shell is in the shape of a cylinder with its axis vertically arranged, and a cylindrical amplification reaction chamber is arranged inside; The partition assembly is used to divide the amplification reaction chamber into three sector-shaped reaction chambers; the three reaction chambers are concentrically arranged; the three reaction chambers are sequentially arranged as a denaturation chamber, a renaturation chamber and an extension chamber along the circumference of the placement shell; the denaturation chamber, the renaturation chamber and the extension chamber are all provided with constant temperature assemblies; the denaturation chamber is provided with a sample loading port, and the renaturation chamber is provided with a sample outlet; the ratio of the arc values ​​of the three reaction chambers to the time required for the sample to stay in the corresponding reaction chamber is the same; There are multiple centrifugal racks, which are arranged in the amplification reaction chamber; the centrifugal racks include a placement plate; the placement plate is arranged in a direction perpendicular to the radial direction of the placement shell; and a plurality of placement holes are arranged on the placement plate; The circulation component is arranged in the amplification reaction chamber, and is used to drive the centrifugal frame to drive the sample to rotate around the axis of the placement shell in the amplification reaction chamber at a uniform speed, and make the sample pass through the denaturation chamber, the renaturation chamber, and the extension chamber in sequence.

2. The integrated controlled gene amplification device according to claim 1, characterized in that: The partition assembly includes three partitions; the partitions are arranged vertically and extend radially along the placement shell; the upper end of the partition is fixed to the upper cavity wall of the amplification reaction chamber, and the lower end is abutted against the lower cavity wall of the amplification reaction chamber; the end of the partition away from the axis of the placement shell is abutted against the side cavity wall of the amplification reaction chamber, and the end close to the axis of the placement shell is fixedly connected with a connecting ring; the connecting ring and the placement shell are coaxially arranged; the upper end of the connecting ring is fixed to the upper cavity wall of the amplification reaction chamber, and the lower end is abutted against the lower cavity wall of the amplification reaction chamber; the partition divides the amplification reaction chamber into a denaturation chamber, a renaturation chamber and an extension chamber.

3. The integrated controlled gene amplification device according to claim 2, characterized in that: The arc value of the denaturation chamber is 90°, the arc value of the renaturation chamber is 90°, and the arc value of the extension chamber is 180°. The sample residence time in the three reaction chambers is set to 30 seconds in the denaturation chamber, 30 seconds in the renaturation chamber, and 1 minute in the extension chamber.

4. The integrated controlled gene amplification device according to claim 3, characterized in that: The centrifugal frame also includes a push plate; two push plates are provided, distributed at both ends of the placement plate along a tangent direction perpendicular to the radial direction of the placement shell; the push plate is fixed to the end of the placement plate; a clamping protrusion is fixed to the end of the push plate away from the placement plate.

5. The integrated controlled gene amplification device according to claim 4, characterized in that: The circulation assembly includes a mounting groove, a rotating disk, a retaining ring, and a guide structure; The mounting groove is an annular groove; the mounting groove is a through groove arranged on the side wall of the connecting ring; the mounting groove and the connecting ring are coaxial; The turntable and the connecting ring are coaxially arranged; the turntable is rotatably installed in the installation groove; the turntable is connected to a servo motor; the servo motor drives the turntable to rotate at a uniform speed; a plurality of card blocks are distributed in an annular manner on the side wall of the turntable; the card blocks are spaced at a preset interval; the preset interval is equal to the length of the placement plate; A clamping groove is provided on the side wall of the clamping block away from the axis of the rotating disk; the clamping groove is in a trapezoidal shape with the small end facing the axis of the rotating disk; The retaining ring is coaxially arranged outside the rotating disk; a circular circulation gap is formed between the retaining ring and the rotating disk; the retaining ring is fixed on the partition plate; a sample loading gap and a sample discharging gap are arranged on the retaining ring; The loading gap corresponds to the loading port, and the discharging gap corresponds to the discharging port; The guiding structure is used to place the centrifugal rack in the amplification reaction chamber, so that the push plates at both ends of the placement plate are stuck between two adjacent blocks, and the corresponding protrusions are stuck in the slots. After the amplification reaction is completed, the centrifugal rack is driven away from the blocks.

6. The integrated controlled gene amplification device according to claim 5, characterized in that: The guide structure includes a sample loading guide rod and a sample discharging guide rod; Two sample loading guide rods are provided, which are horizontally distributed on both sides of the sample loading gap and are arranged in the sample loading port; the sample loading guide rods are arranged along the radial direction of the turntable, and are initially arranged horizontally; one end of the sample loading guide rod close to the axis of the turntable is rotatably mounted on the outer wall of the retaining ring through a horizontally arranged rotating shaft; a sample loading slide groove is provided on the side wall of the sample loading guide rod close to the sample loading gap; the clamping protrusion and the sample loading slide groove are slidably matched; the push plate and the side wall of the sample loading guide rod close to the sample loading gap are abutted and slidably matched; Two sample-discharging guide rods are provided, which are horizontally distributed on both sides of the sample-discharging gap and are arranged in the sample-discharging port; the sample-discharging guide rods are arranged along the radial direction of the turntable, and initially, the end of the sample-discharging guide rod away from the axis of the turntable is higher than the end close to the axis of the turntable; the end of the sample-discharging guide rod close to the axis of the turntable is rotatably mounted on the outer side wall of the retaining ring through a horizontally arranged rotating shaft; a sample-discharging groove is provided on the side wall of the sample-discharging guide rod close to the sample-loading gap; the clamping protrusion and the sample-discharging groove are slidably matched; the push plate and the side wall of the sample-discharging guide rod close to the sample-loading gap are abutted and slidably matched.

7. The integrated controlled gene amplification device according to claim 6, characterized in that: A sample loading motor and a sample discharging motor are fixed on the retaining ring; the output shaft of the sample loading motor is fixedly connected to the rotating shaft of the sample loading guide rod; the output shaft of the sample discharging motor is fixedly connected to the rotating shaft of the sample discharging guide rod.

8. The integrated controlled gene amplification device according to claim 7, characterized in that: A proximity switch is arranged between adjacent card blocks; a controller is arranged in the shell; and the proximity switch and the controller are electrically connected.

9. An integrated controlled gene amplification device according to any one of claims 1 to 8, characterized in that: A sample loading cover is installed on the sample loading port; a sample output cover is installed on the sample output port.

10. The integrated controlled gene amplification device according to claim 9, characterized in that: An upper cover is arranged at the upper end of the placement cavity; an operation screen is arranged at one side of the shell.

Images