A nucleic acid extraction and purification device

Abstract

The present invention discloses a nucleic acid extraction and purification device. The nucleic acid extraction and purification device comprises: a base; a chip mounting mechanism for mounting a microfluidic chip and being movably disposed on the base in the left-hand direction; an extraction drive mechanism comprising an extraction drive pin capable of engaging with the left-hand rotary piston of the microfluidic chip to drive its rotation; a translation drive mechanism capable of movably disposed on the chip mounting mechanism in the left-hand direction and comprising a translation drive member for engaging with the translation piston of the microfluidic chip to drive its movement; and a magnet assembly for applying a magnetic field to magnetic beads within the microfluidic chip. When the left-hand rotary piston engages the extraction drive pin, the magnet assembly has at least a first state and a second state. In the first state, a purification and separation chamber is located within the magnetic field of the magnet assembly; in the second state, the purification and separation chamber is separated from the magnetic field of the magnet assembly. The present invention can automatically extract and purify nucleic acids.

Description

Technical Field

[0001] The invention belongs to the technical field of nucleic acid extraction and relates to a nucleic acid extraction and purification device. Background Art

[0002] At present, microfluidic chips are used in the field of biological detection. They can be placed in a PCR instrument to carry out reactions to achieve the purpose of detection, for example, to extract and purify nucleic acids. In order to avoid contamination, the lysis solution, rinse solution and other reagents required for nucleic acid extraction are pre-stored in the various reagent chambers of the microfluidic chip, and the magnetic beads used to adsorb nucleic acids are also pre-placed in the chambers of the microfluidic chip. When performing nucleic acid extraction, the reagents and samples need to be mixed in order and subjected to lysis, separation, purification reactions, etc. After each reaction, the magnet is close to the purification and separation chamber, and a magnetic field is applied to the magnetic beads therein. The nucleic acids are adsorbed on the magnetic beads, and then the rotary piston is rotated to connect to the waste liquid chamber, and the waste liquid is driven into the waste liquid chamber by the translation piston. Accordingly, it is necessary to design a device that can automatically extract and purify nucleic acids. Summary of the Invention

[0003] In view of the above technical problems, the present invention provides a nucleic acid extraction and purification device that can automatically extract and purify nucleic acids.

[0004] In order to achieve the above object, the present invention adopts the following technical solutions:

[0005] A nucleic acid extraction and purification device, comprising:

[0006] base;

[0007] A chip mounting mechanism, which is used to mount a microfluidic chip, and the chip mounting mechanism is movably disposed on the base in a left-right direction;

[0008] an extraction drive mechanism, located on the left side of the chip mounting mechanism, the extraction drive mechanism comprising an extraction drive pin capable of engaging with the left rotary piston of the microfluidic chip to drive the left rotary piston to rotate;

[0009] a translation drive mechanism, which is movably disposed on the chip mounting mechanism in the left-right direction, the translation drive mechanism comprising a translation drive member for engaging with a translation piston of the microfluidic chip to drive the same to move; and

[0010] a magnet assembly, which is used to apply a magnetic field to the magnetic beads in the purification and separation chamber of the microfluidic chip;

[0011] When the left rotary piston engages the extraction drive pin, the magnet assembly has at least a first state and a second state. In the first state, the purification and separation chamber is located in the magnetic field of the magnet assembly; in the second state, the purification and separation chamber is out of the magnetic field of the magnet assembly.

[0012] According to a preferred embodiment, the magnet assembly is located on the left side of the chip mounting mechanism.

[0013] More preferably, the magnet assembly includes a mounting seat arranged on the base or the extraction drive mechanism, a permanent magnet movably arranged on the mounting seat in the left and right directions, and an electromagnet for driving the permanent magnet to move, and the electromagnet is arranged on or in the mounting seat.

[0014] Furthermore, the chip mounting mechanism includes a chip shell having a chip slot for accommodating the microfluidic chip, and a first notch for the permanent magnet to enter is provided on the left wall of the chip shell.

[0015] According to a preferred embodiment, the chip mechanism includes a chip shell having a chip slot for accommodating the microfluidic chip, and the translation drive component is located on the right side of the chip shell.

[0016] More preferably, a second notch corresponding to the translation driving member is provided on the right wall of the chip housing, and the translation driving member has an initial position located in the second notch.

[0017] According to a preferred embodiment, the translation drive member has a slot for inserting the translation piston, and the slot has an upward-facing notch.

[0018] According to a preferred embodiment, the translation drive mechanism includes a guide rail extending in the left-right direction and provided on the chip mounting mechanism, and a motor for driving the translation drive member to move, and the translation drive member is slidably provided on the guide rail.

[0019] According to a preferred embodiment, the extraction drive mechanism includes a motor and a worm gear transmission mechanism for connecting the motor and the extraction drive pin.

[0020] According to a preferred embodiment, the chip mounting mechanism is movably arranged on the base along the left-right direction through a moving mechanism, and the base is provided with a horizontal guide rail extending along the left-right direction, and the moving mechanism includes a slide movably arranged on the horizontal guide rail along the left-right direction; and / or, the chip mounting mechanism includes a chip shell, a chip slot for accommodating a microfluidic chip is formed in the chip shell, and a through hole for the extraction drive pin to pass through is opened on the left wall of the chip shell.

[0021] According to a preferred embodiment, the nucleic acid extraction and purification device further comprises an amplification drive mechanism located on the right side of the chip mounting mechanism, wherein the amplification drive mechanism comprises an amplification drive pin for engaging with the right rotary piston of the microfluidic chip to drive the same to rotate;

[0022] More preferably, the amplification drive mechanism includes a motor and a worm gear transmission mechanism for connecting the motor and the amplification drive pin.

[0023] More preferably, when viewed from above, the extraction drive mechanism and the amplification drive mechanism are centrally symmetrical.

[0024] According to a preferred embodiment, the moving mechanism further includes a vertical guide rail provided on the slide, and the chip mounting mechanism is movably provided on the vertical guide rail in an up-down direction.

[0025] Furthermore, the nucleic acid extraction and purification device also includes a heating mechanism for heating the microfluidic chip, the heating mechanism includes a heating component, the heating component has a first heating area and a second heating area located below the first heating area, the temperatures of the first heating area and the second heating area are different, and the chip mounting mechanism has a position that can be attached to the first heating area and a position that can be attached to the second heating area.

[0026] According to a preferred embodiment, the heating mechanism is located at the rear side of the chip mounting mechanism, and the heating component is movably disposed on the base so as to be able to move close to or away from the microfluidic chip.

[0027] According to a preferred embodiment, the heating component is rotatably disposed on the base via a rotating shaft, and the axis of the rotating shaft extends in the left-right direction.

[0028] More preferably, the heating mechanism also includes a fixed seat, a rocking arm, a connecting rod, a power source and a connecting piece that can be driven by the power source to move in the left and right directions, the fixed seat is arranged on the base, the rocking arm is rotatably connected to the fixed seat through a first rotating shaft, the power source is arranged on the rocking arm, the connecting piece is rotatably connected to one end of the connecting rod through a second rotating shaft, the other end of the connecting rod is rotatably connected to the fixed seat through a third rotating shaft, and the heating component is arranged on the connecting rod.

[0029] Furthermore, the power source includes a linear motor, and the connecting member is arranged on the output shaft of the linear motor; and / or, the axis lines of the first rotating shaft, the second rotating shaft and the third rotating shaft extend in the left and right directions respectively and are parallel to each other but do not overlap; and / or, the heating mechanism also includes a support plate, which is rotatably connected to the fixed seat through a fourth rotating shaft, the axis lines of the fourth rotating shaft and the third rotating shaft overlap with each other, and the heating component is arranged on the support plate and the connecting rod.

[0030] The principle of nucleic acid extraction and purification is as follows: the sample is loaded into the purification and separation chamber of the microfluidic chip; the microfluidic chip is loaded onto the chip mounting mechanism, and when the microfluidic chip is inserted, its translation piston engages with the translation drive member of the translation drive mechanism; the chip mounting mechanism moves to the left, so that the extraction drive pin engages with the left rotary piston of the microfluidic chip, and in turn drives the left rotary piston to rotate a certain angle, so as to connect the chamber storing the lysate and rinse solution with the purification and separation chamber in turn; the translation drive member of the translation drive mechanism drives the translation piston to move left and right to apply positive or negative pressure to the lysate and other extraction reagents, so that it flows The sample is placed in the purification and separation chamber for reaction treatment; after each reaction, the magnet assembly applies a magnetic field to the purification and separation chamber, and the magnetic beads in the purification and separation chamber are magnetized, thereby adsorbing the lysed or rinsed nucleic acids on them; the extraction drive mechanism drives the left rotary piston to rotate a set angle to connect the purification and separation chamber and the waste liquid chamber, and the translation drive member drives the translation piston to move, discharges the waste liquid in the purification and separation chamber into the waste liquid chamber, and stops applying the magnetic field to the purification and separation chamber; repeat the above steps, pass the rinse liquid into the purification and separation chamber, elute the nucleic acid on the magnetic beads, and use it for subsequent amplification reactions.

[0031] The present invention adopts the above solution and has the following advantages compared with the prior art:

[0032] The nucleic acid extraction and purification device of the present invention has an extraction drive mechanism capable of engaging with the left rotary piston of the microfluidic chip, a translation drive mechanism capable of engaging with the translation piston of the microfluidic chip, and a magnet assembly capable of applying a magnetic field to the magnetic beads of the microfluidic chip to cause them to adsorb nucleic acids for separation and rinsing and purification. Reagents for nucleic acid extraction are mixed with samples in a time sequence for reaction and separation. After the microfluidic chip is loaded, nucleic acid extraction and purification can be automatically performed without the need for other manual operations, which is quick and convenient and can effectively avoid contamination and human errors. BRIEF DESCRIPTION OF THE DRAWINGS

[0033] In order to more clearly illustrate the technical solution of the present invention, the following is a brief introduction to the drawings required for use in the description of the embodiments. 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 any creative work.

[0034] Figure 1 is a three-dimensional structural diagram of a nucleic acid extraction and purification device according to an embodiment of the present invention, wherein the cover is not shown;

[0035] Figure 2 for Figure 1 A front view of the nucleic acid extraction and purification device shown;

[0036] Figure 3 for Figure 1A top view of the nucleic acid extraction and purification device shown;

[0037] Figure 4a and Figure 4b These are schematic diagrams of the structure of the microfluidic chip from two different perspectives;

[0038] Figure 5 It is a structural diagram of the mobile mechanism;

[0039] Figure 6 A three-dimensional structural diagram of the chip mounting mechanism, translation drive mechanism, and moving mechanism;

[0040] Figure 7 for Figure 6 a side view of the mechanism shown;

[0041] Figure 8 A three-dimensional diagram of the chip mounting mechanism and the translation drive mechanism;

[0042] Figure 9 A side view of the extraction drive mechanism;

[0043] Figure 10 for Figure 9 Cross-sectional view along the AA axis;

[0044] Figure 11 It is a three-dimensional structural diagram of the heating mechanism;

[0045] Figure 12 It is a front view of the heating mechanism;

[0046] Figure 13 is a side view of the heating mechanism;

[0047] Figure 14 is a top view of the heating mechanism;

[0048] Figure 15 and Figure 16 They are structural schematic diagrams of the optical detection mechanism from two different perspectives;

[0049] Figure 17 A three-dimensional diagram of the magnet assembly;

[0050] Figure 18 A top view of the magnet assembly.

[0051] in,

[0052] 1. Base; 10. Bottom plate; 11. Mounting plate;

[0053] 2. Chip mounting mechanism; 20. Chip housing; 200. Chip slot; 201. Bottom wall; 202. Front wall; 203. Left wall; 2031. Folding edge; 2032. Through hole; 2033. Notch; 204. Right wall; 2041. Folding edge; 2042. Through hole; 2043. Notch; 205. Hollow portion;

[0054] 3. Extraction drive mechanism; 31. Extraction drive pin; 32. Motor; 33. Worm gear transmission mechanism; 34. Reducer;

[0055] 4. Amplify the drive mechanism; 41. Extract the drive pin;

[0056] 5. Heating mechanism; 50. Heating assembly; 500. Radiator; 502. Heat conducting plate; 503. Cooling fan; 504. Insulation foam; 51. Fixing seat; 52. Rocking arm; 53. Connecting rod; 54. Linear motor; 55. Connecting member; 56. Support plate; a. First rotating shaft; b. Second rotating shaft; c. Third rotating shaft; d. Fourth rotating shaft;

[0057] 6. Fluorescence detection mechanism; 60. Detection unit; 61. Horizontal guide rail; 62. Motor;

[0058] 7. Moving mechanism; 70. Horizontal guide rail; 71. Slide; 72. Vertical guide rail; 73. First chip motor; 74. Second chip motor; 740. Screw;

[0059] 8. Translation drive mechanism; 80. Translation drive member; 801. Slot; 81. Guide rail; 82. Motor; 820. Screw;

[0060] 9. Magnet assembly: 90. Mounting seat; 91. Mounting shaft; 92. Permanent magnet; 93. Sensor sheet; 94. Proximity switch;

[0061] 100. Microfluidic chip; 101. Left rotary piston; 102. Right rotary piston; 103. Translational piston; 104. Separation area; 105. Lower part; 106. Step surface. DETAILED DESCRIPTION

[0062] The preferred embodiments of the present invention are described in detail below with reference to the accompanying drawings so that the advantages and features of the present invention can be more easily understood by those skilled in the art. It should be noted that the description of these embodiments is intended to help understand the present invention, but does not constitute a limitation of the present invention.

[0063] In the description of this application, it should be understood that the terms "upper", "lower", "left", "right", "front", "back", "vertical", "horizontal", "inside" and "outside" indicate positions or positional relationships based on the positions or positional relationships shown in the accompanying drawings. They are only for the convenience of describing this application and simplifying the description, and do not indicate or imply that the device or element referred to must have a specific orientation, be constructed and operate in a specific orientation. Therefore, they should not be understood as limiting this application. Figure 2 For example, left and right correspond to the left and right sides of the paper in the figure, up and down correspond to the lower and upper sides of the paper in the figure, and front and back correspond to the direction perpendicular to the paper and the side closer to the observer is the front. Figure 2 The viewing angles shown are for reference only.

[0064] Figures 1 to 3 The present invention shows a nucleic acid extraction and purification device according to an embodiment of the present invention, which is adapted to a microfluidic chip to extract and purify nucleic acids. Figures 1 to 3 As shown, the nucleic acid extraction and purification device includes a base 1 and a chip mounting mechanism 2, an extraction drive mechanism 3, a moving mechanism 7, a translation drive mechanism 8 and a magnet assembly 9, etc., which are arranged on the base 1. The nucleic acid extraction and purification device can further perform PCR amplification and subsequent fluorescence detection on the extracted and purified nucleic acid. Correspondingly, the nucleic acid extraction and purification device also includes an amplification drive mechanism 4, a heating mechanism 5 and a fluorescence detection mechanism 6. The nucleic acid extraction and purification device also includes a cover body (not shown in the figure) provided on the base 1 for covering the above-mentioned various mechanisms and components. A slit-shaped through hole for the entry and exit of the microfluidic chip is provided on the upper side wall of the cover body, and the slit-shaped through hole is located directly above the chip mounting mechanism 2.

[0065] The structure of the microfluidic chip 100 is as follows: Figure 4a and Figure 4bAs shown, the microfluidic chip 100 has multiple chambers, and microchannels for liquid to pass through are provided between the chambers; the microfluidic chip 100 includes a left rotary piston 101 and a right rotary piston 102, which are used to switch the connectivity of the microchannels. By rotating the left rotary piston 101 or the right rotary piston 102, some microchannels can be connected and other microchannels can be cut off; the microfluidic chip 100 also includes a translation piston 103, which is used to provide a driving force for the flow of liquid. As the translation piston 103 moves, negative pressure or positive pressure can be applied to the liquid to drive it to the specified chamber. The microfluidic chip 100 in this embodiment is a vertical microfluidic chip 100, and its dimensions (length) in the left-right direction and the dimensions (height) in the up-down direction are respectively greater than its dimensions (width) in the front-back direction. The left rotary piston 101 is higher than the right rotary piston 102. Both are inserted into the chip body of the microfluidic chip 100 and can rotate around their own axes. The left end of the left rotary piston 101 is exposed to facilitate engagement with the extraction drive mechanism 3, which is mainly used to control the on-off switching of the chambers storing nucleic acid extraction and purification reagents and the purification and separation chambers. The right end of the right rotary piston 102 is exposed to facilitate engagement with the amplification drive mechanism 4, which is mainly used to control the on-off switching between the amplification reaction chamber and the amplification detection chambers. The translation piston 103 is inserted into the chip body of the microfluidic chip 100 and can move left and right. Its left end extends outside the chip body of the microfluidic chip 100 to facilitate engagement with the translation drive mechanism 8. The upper portion of the microfluidic chip 100 is provided with a purification and separation chamber containing magnetic beads. Accordingly, the left side surface of the body of the microfluidic chip 100 is provided with a separation area 104 that can contact and cooperate with the magnet assembly 9. The lower portion 105 of the microfluidic chip 100 is equipped with multiple amplification detection cavities arranged side by side along the left-right direction. The portions of the chip body corresponding to the amplification detection cavities are transparent or translucent to allow light to enter and excitation light to exit. The lower portion 105 of the microfluidic chip 100 is thinner than the upper portion and has a downward-facing step surface 106.

[0066] The chip mounting mechanism 2 is used to mount the microfluidic chip 100, which is arranged on the base 1 so as to be movable in the left and right directions. Figure 3 、 Figure 5 and Figure 6As shown, the chip mounting mechanism 2 is arranged on the base 1 via a moving mechanism 7, and can move relative to the base 1 in the left-right direction and the up-down direction. The moving mechanism 7 includes a slide 71 connected to the base 1 so as to be movable in the left-right direction, and the chip mounting mechanism 2 is arranged on the slide 71 so as to be movable in the up-down direction. Specifically, the base 1 includes a base plate 10 and a plurality of upwardly extending mounting plates 11 fixed to the base plate 10. One of the mounting plates 11 is provided with a horizontal guide rail 70 extending in the left-right direction, and the slide 71 is slidably arranged on the horizontal guide rail 70; the slide 71 is provided with a vertical guide rail 72 extending in the up-down direction, and the chip mounting mechanism 2 is slidably arranged on the vertical guide rail 72. The moving mechanism 7 also includes a first chip motor 73 for driving the slide 71 to move along the horizontal guide rail 70, and the slide 71 is connected to the output shaft of the first chip motor 73 through a screw rod; the moving mechanism 7 also includes a second chip motor 74 for driving the chip mounting mechanism 2 to move in the up and down directions, and the chip mounting mechanism 2 is connected to the output shaft of the second chip motor 74 through a screw rod 740, and the screw rod 740 extends in the up and down directions and is rotatable around its own axis and is arranged on the mounting plate 11.

[0067] Combine Figures 6 to 8As shown, the chip mounting mechanism 2 includes a chip housing 20, within which is formed a chip slot 200 for accommodating the microfluidic chip 100. The chip housing 20 includes a bottom wall 201, a front wall 202, a left wall 203, and a right wall 204. The four walls define the chip slot 200 for inserting the microfluidic chip 100. The rear side of the left wall 203 has a folded edge 2031 that bends and extends to the right, and the rear side of the right wall 204 has a folded edge 2041 that bends and extends to the left. The folded edges 2031 and 2041 serve to limit the microfluidic chip 100, thereby preventing the microfluidic chip 100 from falling out of the rear side of the chip slot 200. The rear edge of the bottom wall 201 is located a distance in front of the folded edge 2031 of the left wall 203 and the folded edge 2041 of the right wall 204, thereby forming a hollow portion 205 for the lower portion of the microfluidic chip 100 to pass downward. The left wall 203 has a through hole 2032 and a notch 2033 at the top; the right wall 204 has a through hole 2042 and a notch 2043 at the top. When the microfluidic chip 100 is loaded into the chip shell 20, the step surface 106 rests on the bottom wall 201, and the lower part of the microfluidic chip 100 passes through the chip shell 20 and is located below the chip shell 20, so that the fluorescence detection mechanism 6 can illuminate the amplification detection cavity at the lower part of the microfluidic chip 100 and collect fluorescence; the left end of the left rotary piston 101 of the microfluidic chip 100 faces the through hole 2032 on the left wall 203, and the left end of the translation piston 103 is located in the notch 2043 on the right wall 204; the right end of the right rotary piston 102 faces the through hole 2042 on the right wall 204, and the separation area 104 faces the notch 2033 on the left wall 203; the area to be heated of the microfluidic chip 100 faces the rear side and is not blocked by the chip shell 20, and can be in direct contact with the heating mechanism 5.

[0068] like Figures 1 to 3 、 Figure 9 、 Figure 10 As shown, the extraction drive mechanism 3 is located on the left side of the chip mounting mechanism 2. The extraction drive mechanism 3 includes an extraction drive pin 31 for engaging with the left rotary piston 101 of the microfluidic chip 100 to drive its rotation. The amplification drive mechanism 4 is located on the right side of the chip mounting mechanism 2. The amplification drive mechanism 4 includes an amplification drive pin 41 for engaging with the right rotary piston 102 of the microfluidic chip 100 to drive its rotation. The chip mounting mechanism 2 at least has a position for engaging the extraction drive pin 31 and disengaging the amplification drive pin 41 for the microfluidic chip 100, and a position for engaging the amplification drive pin 41 and disengaging the extraction drive pin 31 for the microfluidic chip 100.

[0069] In a top view, the extraction drive mechanism 3 and the amplification drive mechanism 4 are centrally symmetrical. Figure 9 and Figure 10The extraction drive mechanism 3 is described in detail. The amplification drive mechanism 4 is similar to the extraction drive mechanism 3. Figure 9 and Figure 10 The extraction drive mechanism 3 includes a motor 32 arranged on the base 1 and a worm gear transmission mechanism 33 for connecting the motor 32 and the extraction drive pin 31. The worm gear transmission mechanism 33 is arranged in a reduction box 34, and the reduction box 34 is fixedly arranged on the bottom plate 10 of the base 1. Specifically, the output shaft of the motor 32 extends in the front-to-back direction, and the axis of the extraction drive pin 31 extends in the left-to-right direction. After the motor 32 is running, the torque output by the motor 32 is changed in direction by the worm gear transmission mechanism 33 to drive the extraction drive pin 31 to rotate, which can then drive the left rotary piston 101 of the microfluidic chip 100 to rotate. The amplification drive mechanism 4 also includes a motor and a worm gear transmission mechanism. The working principle is similar to that of the extraction drive mechanism 3 and will not be elaborated here.

[0070] The heating mechanism 5 is located at the rear side of the chip mounting mechanism 2 to heat the microfluidic chip 100. Figures 11 to 14 As shown, the heating mechanism 5 includes a heating component 50 that is movably mounted on the base 1 and can be moved closer to or further away from the microfluidic chip 100. The heating component 50 has a first heating area and a second heating area located below the first heating area. The first heating area and the second heating area have different temperatures. The chip mounting mechanism 2 moves in the vertical direction so that it can fit in different heating areas.

[0071] The heating assembly 50 is rotatably mounted on the base 1 via a rotating shaft, the axis of which extends in the left-right direction. Specifically, the heating mechanism 5 further includes a fixed base 51, a rocking arm 52, a connecting rod 53, a power source, and a connecting member 55 capable of being driven in the left-right direction by the power source. The fixed base 51 is mounted on the base 1, the rocking arm 52 is rotatably connected to the fixed base 51 via a first rotating shaft a, the power source is mounted on the rocking arm 52, the connecting member 55 is rotatably connected to one end of the connecting rod 53 via a second rotating shaft b, and the other end of the connecting rod 53 is rotatably connected to the fixed base 51 via a third rotating shaft c. The heating assembly 50 is mounted on the connecting rod 53. The power source includes a linear motor 54, and the connecting member 55 is mounted on the output shaft of the linear motor 54. The axis of the first rotating shaft a, the second rotating shaft b, and the third rotating shaft c extend in the left-right direction and are parallel to each other but do not overlap. The heating mechanism 5 also includes a support plate 56, which is rotatably connected to the fixed base 51 via a fourth rotation axis d. The axis of the fourth rotation axis d and the third rotation axis c coincide with each other. The heating assembly 50 is mounted on the support plate 56 and the connecting rod 53. The heating assembly 50 swings in this manner, allowing it to attach to or detach from the microfluidic chip 100, eliminating the need for excessive space and resulting in a compact structure. Furthermore, the movable connection, such as the connecting rod 53, prevents excessive frictional losses between components.

[0072] The heating assembly 50 specifically includes a radiator 500, a plurality of heating elements (not shown in the figure) provided on the front side of the radiator 500, a heat conducting plate 502 respectively covering each heating element, and a cooling fan 503 provided on the rear side of the radiator 500. The plurality of heating elements are spaced apart in the vertical direction. Correspondingly, the plurality of heat conducting plates 502 are also spaced apart in the vertical direction, wherein a first heating area is formed on one of the heat conducting plates 502, and a second heating area is formed on another heat conducting plate 502 below the heat conducting plate 502. The heating element is specifically a TEC cooling plate, which is embedded directly behind the heat conducting plate 502. The heating assembly 50 also includes heat insulation cotton provided around the heating element. Each heating element can have different temperatures. By moving the chip mounting mechanism 2 up and down, the amplification reaction can be circulated at different temperatures according to the set reaction program.

[0073] The fluorescence detection mechanism 6 is located at the lower side of the chip mounting mechanism 2 and is used to fit or be close to the lower portion 105 of the microfluidic chip 100 to illuminate the amplification detection cavity and collect excitation light. Figure 15 and Figure 16 As shown, the fluorescence detection mechanism 6 is movably disposed on the base 1 in the left-right direction. Specifically, a horizontal guide rail 61 extending in the left-right direction is provided on the bottom plate 10 of the base 1, and the fluorescence detection mechanism 6 is slidably disposed on the horizontal guide rail 61. The bottom plate 10 is also provided with a motor 62 for driving the fluorescence detection mechanism 6 to move left and right. The motor 62 is specifically connected to the fluorescence detection mechanism 6 via a screw. By moving the fluorescence detection mechanism 6, a single fluorescence detection mechanism 6 can complete the detection of all amplification detection cavities of the microfluidic chip 100. For example, in one specific embodiment, the fluorescence detection mechanism 6 has four detection units 60 corresponding to four different colors of excitation light. The multiple detection units 60 are sequentially arranged in parallel in the left-right direction, and the microfluidic chip 100 has 12 amplification detection cavities arranged in parallel in the left-right direction. The fluorescence detection mechanism 6 is first aligned with amplification detection cavities 1 to 4. After the detection is completed, the fluorescence detection mechanism 6 is moved to align with amplification detection cavities 5 to 8. After the detection is completed, the fluorescence detection mechanism 6 is again moved to align with amplification detection cavities 9 to 12, thereby completing the detection of all amplification detection cavities.

[0074] Combine Figures 6 to 8As shown, the translation drive mechanism 8 is disposed on the chip mounting mechanism 2. The translation drive mechanism 8 includes a translation drive member 80 for engaging with the translation piston 103 of the microfluidic chip 100 to drive its movement. The translation drive member 80 is disposed on the chip mounting mechanism 2 so as to be movable in the left-right direction. The translation drive member 80 is located on the right side of the chip mounting mechanism 2. The translation drive member 80 can move left-right along with the chip mounting mechanism 2 and can also move left-right relative to the chip mounting mechanism 2 to drive the translation piston 103 to slide left-right within the chip body of the microfluidic chip 100. The translation drive member 80 has a slot 801 for inserting the translation piston 103, and the slot 801 has a notch facing upward; when the translation drive member 80 is in its initial position, it is embedded in the notch 2043 of the right wall 204 of the chip shell 20. When the microfluidic chip 100 is inserted into the chip shell 20, the right end of its translation piston 103 falls from top to bottom into the slot 801 of the translation drive member 80 and engages with the translation drive member 80. During the entire detection process, the translation drive member 80 and the translation piston 103 always remain engaged with each other.

[0075] The translation drive mechanism 8 also includes a guide rail 81 and a motor 82. The guide rail 81 extends left and right and is mounted on the chip mounting mechanism 2. Specifically, it is fixedly connected to the chip housing 20. The translation drive member 80 is slidably mounted on the guide rail 81. A motor 82 is mounted on the chip mounting mechanism 2 and is used to drive the translation drive member 80 to slide left and right. Specifically, the motor is connected to the translation drive member 80 via a screw 820.

[0076] The magnet assembly 9 is used to apply a magnetic field to the magnetic beads in the purification and separation chamber of the microfluidic chip 100. When the left rotary piston 101 of the microfluidic chip 100 is engaged with the extraction drive pin 31, the magnet assembly 9 has at least a first state and a second state. In the first state, the purification and separation chamber is located in the magnetic field of the magnet assembly 9; in the second state, the purification and separation chamber is out of the magnetic field of the magnet assembly 9. Figure 17 and Figure 18As shown, the magnet assembly 9 includes a mounting seat 90 arranged on the base 1, a permanent magnet 92 movably arranged on the mounting seat 90 and an electromagnet (not shown in the figure) for driving the permanent magnet 92 to move. The electromagnet is arranged on or in the mounting seat 90. Specifically, the mounting seat 90 is fixedly arranged on the reduction gearbox 30 of the extraction drive mechanism 3, and the permanent magnet 92 is movably arranged on the mounting seat 90 through a mounting shaft 91. The mounting shaft 91 extends in the left and right directions and is connected to the mounting seat 90 movably in the left and right directions. The permanent magnet 92 is fixedly arranged at the left end of the mounting shaft 91 and faces the right side; the electromagnet is arranged in the mounting seat 90 to drive the mounting shaft 91 to move. When the electromagnet is energized, the mounting shaft 91 extends to the right to drive the permanent magnet 92 to move and adhere to the separation area 104 of the microfluidic chip 100, applying a magnetic field to the purification and separation chamber, and the magnet assembly 9 is in the first state; when the electromagnet is de-energized, the mounting shaft 91 retracts to the left, the permanent magnet 92 disengages from the separation area 104 of the microfluidic chip 100, and its magnetic field leaves the purification and separation chamber, and the magnet assembly 9 is in the second state. A sensing member 93 is also provided on the mounting shaft 91, and a proximity switch 94 for detecting the position of the permanent magnet 92 is provided on the mounting base 90. The proximity switch 94 is specifically a photoelectric sensor. When the sensing member 93 enters or leaves the detection area of ​​the photoelectric sensor, the photoelectric sensor is triggered.

[0077] The working process of the nucleic acid extraction and purification device is as follows:

[0078] Insert the microfluidic chip 100 into the chip slot 200. After insertion, the right end of the translation piston 103 falls into the slot 801 of the translation driver 80, so that the two are engaged.

[0079] When it is necessary to rotate the left rotary piston 101 to connect certain chambers, the chip mounting mechanism 2 is moved to the left as a whole, so that the extraction drive pin 31 and the left end of the left rotary piston 101 are engaged (specifically, the extraction drive pin 31 passes through the through hole 2032 on the chip shell 20 and is inserted into the cross groove on the left end of the left rotary piston 101), the motor of the extraction drive mechanism 3 is running, the extraction drive pin 31 rotates and drives the left rotary piston 101 to rotate; at this time, the right rotary piston 102 is disengaged from the microfluidic chip 100; at the same time, the translation drive member 80 and the translation piston 103 are engaged with each other, and when the left rotary piston 101 connects certain chambers, the motor 82 of the third drive member mechanism 8 is running, and the translation drive member 80 moves left and right, thereby driving the translation piston 103 to move left and right in the microfluidic chip 100, thereby providing negative pressure or positive pressure to promote the flow of liquid between connected chambers;

[0080] When it is necessary to rotate the right rotary piston 102 to connect other chambers, the chip mounting mechanism 2 is moved to the right as a whole, so that the amplification drive pin 41 and the right end of the right rotary piston 102 are engaged (specifically, the amplification drive pin 41 passes through the through hole 2032 on the chip shell 20 and is inserted into the cross groove on the right end of the right rotary piston 102), the motor of the amplification drive mechanism 4 is running, the amplification drive pin 41 rotates and drives the right rotary piston 102 to rotate; at this time, the left rotary piston 101 and the permanent magnet 92 are both separated from the microfluidic chip 100; at the same time, the translation drive member 80 and the translation piston 103 are engaged with each other. When the right rotary piston 102 connects certain chambers, the motor 82 of the third drive member mechanism 8 is running, and the translation drive member 80 moves left and right, thereby driving the translation piston 103 to move left and right in the microfluidic chip 100, thereby providing negative pressure or positive pressure to promote the flow of liquid between the connected chambers;

[0081] When it is necessary to adsorb the magnetic beads in the microfluidic chip 100 (specifically located in a purification and separation chamber) to separate the extracted nucleic acid (nucleic acid extraction), the chip mounting mechanism 2 is moved to the left as a whole to the specified position, the electromagnet of the magnet assembly 9 is energized, and the mounting shaft 91 is extended to the right, so that the permanent magnet 92 is attached to the separation area 104 on the left side of the microfluidic chip 100; at the same time, the left rotary piston 101 is engaged with the extraction drive pin 31 and the translation piston 103 is engaged with the translation drive member 80, the left rotary piston 101 is rotated to connect the sample extraction and separation chamber with the waste liquid chamber, and then the translation piston 103 is moved left and right to allow the separated waste liquid to flow into the waste liquid chamber;

[0082] When the microfluidic chip 100 is undergoing an amplification reaction, the motor 54 of the heating mechanism 5 is operated, causing the heating assembly 50 to swing forward and attach to the rear side of the microfluidic chip 100. After attaching, according to a cycle program, the microfluidic chip 100 is attached to the first heating area for a period of time to perform a reaction at the first temperature; the chip mounting mechanism 2 is moved downward, causing the microfluidic chip 100 and the second heating area to attach to each other for a period of time to perform a reaction at the second temperature; the chip mounting mechanism 2 is moved upward, causing the microfluidic chip 100 to attach to the first heating area, and this step is repeated according to the number of cycles.

[0083] When fluorescence needs to be collected for detection, the chip mounting mechanism 2 is moved downward, and the fluorescence detection mechanism 6 faces the amplification detection cavity of the microfluidic chip 100. The fluorescence detection mechanism 6 is moved left and right to complete the detection of all amplification detection cavities.

[0084] The working principle of nucleic acid extraction and purification using this nucleic acid extraction and purification device is generally as follows:

[0085] The microfluidic chip is loaded, and the chip mounting mechanism moves leftward so that the left rotary piston engages the first driving pin;

[0086] Drive the left rotary piston and the translation piston to pass the lysate into the purification and separation chamber to react with the sample therein;

[0087] After the reaction is completed, the magnet assembly applies a magnetic field to the purification and separation chamber, and the lysed nucleic acids are adsorbed on the magnetic beads, driving the left rotary piston and the translation piston to discharge the unadsorbed cell debris and the like into the waste liquid chamber together with the waste liquid;

[0088] The permanent magnet of the magnet assembly retracts, causing its magnetic field to separate from the purification and separation chamber; the left rotary piston is driven to translate the piston, allowing the rinse solution to enter the purification and separation chamber, where the rinse solution washes the nucleic acid off the magnetic beads and rinses them;

[0089] After rinsing, the permanent magnet of the magnet assembly extends to apply a magnetic field to the purification and separation chamber. The rinsed nucleic acids are adsorbed on the magnetic beads, driving the left rotary piston and the translation piston to discharge the unadsorbed waste and other waste into the waste liquid chamber together with the waste liquid.

[0090] The above rinsing steps can be repeated multiple times;

[0091] The permanent magnet of the magnet assembly retracts, causing its magnetic field to separate from the purification and separation chamber; the left rotary piston and translation piston are driven to pass the eluent into the purification and separation chamber, where the eluent elutes the nucleic acid from the magnetic beads for subsequent amplification and detection.

[0092] After the microfluidic chip 100 is loaded, the nucleic acid extraction and purification device can automatically perform nucleic acid extraction, purification, PCR amplification, and detection, eliminating the need for manual operations. This makes detection quick and convenient, effectively avoiding contamination and human error, and providing relatively accurate test results. The device is particularly suitable for nucleic acid extraction, purification, and detection. The various mechanisms of the device are rationally arranged, resulting in a compact structure, making it compact and space-saving.

[0093] As used in this specification and claims, the terms "comprises" and "include" merely indicate the inclusion of the steps and elements specifically identified, and these steps and elements do not constitute an exclusive list; a method or apparatus may also include other steps or elements. The term "and / or" as used herein includes any combination of one or more of the associated listed items.

[0094] It should be noted that, unless otherwise specified, when a feature is referred to as being "fixed" or "connected" to another feature, it may be directly fixed or connected to the other feature or indirectly fixed or connected to the other feature. Furthermore, terms such as "upper," "lower," "left," and "right" used in this disclosure are intended solely to describe the relative positions of the components of the disclosure as shown in the accompanying drawings.

[0095] It will be further understood that the terms "first," "second," and the like are used to describe various types of information, but such information should not be limited to these terms. These terms are used solely to distinguish information of the same type from one another and do not indicate a particular order or level of importance. In fact, the terms "first," "second," and the like are fully interchangeable. For example, first information could be referred to as second information, and similarly, second information could be referred to as first information without departing from the scope of this disclosure.

[0096] The above embodiment is intended only to illustrate the technical concepts and features of the present invention and is a preferred embodiment. Its purpose is to enable those skilled in the art to understand the contents of the present invention and implement them accordingly. It is not intended to limit the scope of protection of the present invention. Any equivalent changes or modifications made based on the principles of the present invention are intended to be covered by the scope of protection of the present invention.

Claims

1. A nucleic acid extraction and purification device, characterized in that: include: base; A chip mounting mechanism, which is used to mount a microfluidic chip, and the chip mounting mechanism is movably disposed on the base in a left-right direction; an extraction drive mechanism, located on the left side of the chip mounting mechanism, the extraction drive mechanism comprising an extraction drive pin capable of engaging with the left rotary piston of the microfluidic chip to drive the left rotary piston to rotate; an amplification drive mechanism located on the right side of the chip mounting mechanism, the amplification drive mechanism comprising an amplification drive pin capable of engaging with the right rotary piston of the microfluidic chip to drive the same to rotate; a translation drive mechanism, which is movably disposed on the chip mounting mechanism in the left-right direction, the translation drive mechanism comprising a translation drive member for engaging with a translation piston of the microfluidic chip to drive the same to move; and a magnet assembly, which is used to apply a magnetic field to the magnetic beads in the purification and separation chamber of the microfluidic chip; When the left rotary piston is engaged with the extraction drive pin, the magnet assembly has at least a first state and a second state. In the first state, the purification and separation chamber is located within the magnetic field of the magnet assembly; in the second state, the purification and separation chamber is out of the magnetic field of the magnet assembly. The chip mounting mechanism is arranged on the base via a moving mechanism and is capable of moving leftward and rightward relative to the base; the chip mounting mechanism has at least a position for causing the microfluidic chip to engage with the extraction drive pin and disengage from the amplification drive pin, and a position for causing the microfluidic chip to engage with the amplification drive pin and disengage from the extraction drive pin; The translation drive member has a slot for inserting the translation piston of the microfluidic chip, and the slot has a notch facing upward; when the microfluidic chip is inserted into the chip mounting mechanism, the right end of the translation piston of the microfluidic chip falls from top to bottom into the slot to engage with the translation drive member.

2. The nucleic acid extraction and purification device according to claim 1, characterized in that: The magnet assembly is located on the left side of the chip mounting mechanism.

3. The nucleic acid extraction and purification device according to claim 2, characterized in that: The magnet assembly includes a mounting seat arranged on the base or the extraction drive mechanism, a permanent magnet movably arranged on the mounting seat in the left and right directions, and an electromagnet for driving the permanent magnet to move, wherein the electromagnet is arranged on or in the mounting seat.

4. The nucleic acid extraction and purification device according to claim 3, characterized in that: The chip mounting mechanism comprises a chip shell, wherein the chip shell has a chip slot for accommodating the microfluidic chip, and a first notch for the permanent magnet to enter is provided on the left wall of the chip shell.

5. The nucleic acid extraction and purification device according to claim 1, characterized in that: The chip mechanism comprises a chip shell, wherein the chip shell has a chip slot for accommodating the microfluidic chip, and the translation driving component is located on the right side of the chip shell.

6. The nucleic acid extraction and purification device according to claim 5, characterized in that: A second notch corresponding to the translation driving member is provided on the right wall of the chip housing, and the translation driving member has an initial position located in the second notch.

7. The nucleic acid extraction and purification device according to claim 1, 5 or 6, characterized in that: The translation driving mechanism includes a guide rail provided on the chip mounting mechanism and extending in the left-right direction and a motor for driving the translation driving member to move. The translation driving member is slidably provided on the guide rail.

8. The nucleic acid extraction and purification device according to claim 1, characterized in that: The extraction drive mechanism includes a motor and a worm gear transmission mechanism for connecting the motor and the extraction drive pin.

9. The nucleic acid extraction and purification device according to claim 1, characterized in that: The base is provided with a horizontal guide rail extending in the left-right direction, and the moving mechanism includes a slide seat that can be movably arranged on the horizontal guide rail in the left-right direction; and / or, the chip mounting mechanism includes a chip shell, a chip slot for accommodating a microfluidic chip is formed in the chip shell, and a through hole for the extraction drive pin to pass through is opened on the left wall of the chip shell.

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