A nucleic acid extraction and amplification device

Abstract

This invention discloses a nucleic acid extraction and amplification device. The device includes: a chip mounting mechanism for mounting a microfluidic chip, the chip mounting mechanism being movable in a left-right direction; a first driving mechanism located on the left side of the chip mounting mechanism, the first driving mechanism including a first driving pin for engaging with a left-side rotating piston of the microfluidic chip to drive its rotation; and a second driving mechanism located on the right side of the chip mounting mechanism, the second driving mechanism including a second driving pin for engaging with a right-side rotating piston of the microfluidic chip to drive its rotation; the chip mounting mechanism has at least a first position for engaging the microfluidic chip with the first driving pin and disengaging it from the second driving pin, and a second position for engaging the microfluidic chip with the second driving pin and disengaging it from the first driving pin. This invention enables automated nucleic acid extraction and amplification.

Description

TECHNICAL FIELD

[0001] The present application belongs to the field of nucleic acid extraction and amplification technology, and relates to a nucleic acid extraction and amplification device. BACKGROUND

[0002] At present, microfluidic chips are applied in the field of biological detection, which can be put into a PCR instrument for reaction to achieve the purpose of detection, such as nucleic acid extraction and amplification. In order to avoid pollution, the reagents required for the reaction are put into the chambers in the microfluidic chip in advance. During the reaction, the liquid flow direction is controlled according to the set reaction program, and the reagents, samples, reaction liquids and the like are flowed into the specified chambers. Therefore, a plurality of pistons for controlling the liquid flow direction or the communication between the chambers or not are arranged on the microfluidic chip. By rotating or moving these pistons, the communication state of the flow channels between the chambers can be switched, or the driving force for controlling the liquid flow can be provided. Accordingly, a driving mechanism needs to be arranged in the nucleic acid extraction and amplification device, which can automatically perform nucleic acid extraction and amplification. SUMMARY

[0003] In view of the above technical problems, the present application provides a nucleic acid extraction and amplification device which can automatically perform nucleic acid extraction and amplification.

[0004] To achieve the above purpose, the present application adopts the following technical scheme:

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

[0006] A chip mounting mechanism for mounting a microfluidic chip, the chip mounting mechanism being movable in left-right direction;

[0007] A first driving mechanism located at the left side of the chip mounting mechanism, the first driving mechanism comprising a first driving pin for engaging with a left side rotary piston of the microfluidic chip to drive the rotary piston to rotate; and

[0008] A second driving mechanism located at the right side of the chip mounting mechanism, the second driving mechanism comprising a second driving pin for engaging with a right side rotary piston of the microfluidic chip to drive the rotary piston to rotate.

[0009] The chip mounting mechanism has at least a first position for engaging the microfluidic chip with the first driving pin and disengaging the microfluidic chip from the second driving pin, and a second position for engaging the microfluidic chip with the second driving pin and disengaging the microfluidic chip from the first driving pin.

[0010] According to a preferred embodiment, the first driving mechanism or the second driving mechanism comprises a motor and a worm gear transmission mechanism for connecting the motor and the first driving pin or the second driving pin.

[0011] More preferably, from a top-down view, the first drive mechanism and the second drive mechanism are centrally symmetrical.

[0012] According to a preferred embodiment, the chip mounting mechanism is movable on a base in a left-right direction via a moving mechanism. The base is provided with a horizontal guide rail extending in a left-right direction, and the moving mechanism includes a slide seat that is movable on the horizontal guide rail in a left-right direction.

[0013] More preferably, the moving mechanism further includes a vertical guide rail disposed on the slide, and the chip mounting mechanism is movably disposed on the vertical guide rail in the up-down direction.

[0014] Furthermore, the nucleic acid extraction and amplification device also includes a heating mechanism for heating the microfluidic chip. The heating mechanism includes a heating component, which has a first heating region and a second heating region located below the first heating region. The first heating region and the second heating region have different temperatures. The chip mounting mechanism has a position that can fit with the first heating region and a position that can fit with the second heating region.

[0015] According to a preferred embodiment, the nucleic acid extraction and amplification device further includes a third driving mechanism, which includes a translational driving member for engaging with a translational piston of the microfluidic chip to drive its movement. The translational driving member is movably disposed on the chip mounting mechanism in a left-right direction, and the translational driving member is located on the left or right side of the chip mounting mechanism.

[0016] More preferably, the translation drive has a slot for inserting the translation piston, the slot having an upward-facing opening; the third drive mechanism includes a guide rail extending in the left-right direction on the chip mounting mechanism and a motor for driving the translation drive to move, the translation drive being slidably disposed on the guide rail.

[0017] Furthermore, the chip mounting mechanism includes a chip housing, in which a chip groove for accommodating a microfluidic chip is formed, and a notch is provided on the left or right side wall of the chip housing to cooperate with the translation drive, and the translation drive has an initial position embedded in the notch.

[0018] According to a preferred embodiment, the chip mounting mechanism includes a chip housing, in which a chip groove for accommodating a microfluidic chip is formed, a through hole is provided on the left wall of the chip housing for the first driving pin to pass through, and a through hole is provided on the right wall of the chip housing for the second driving pin to pass through.

[0019] According to a preferred embodiment, the heating mechanism is located on the rear side of the chip mounting mechanism, and the heating component is movably disposed on the base so as to be close to or away from the microfluidic chip.

[0020] According to a preferred embodiment, the heating component is rotatably mounted on the base via a rotating shaft, the axis of which extends in the left-right direction.

[0021] More preferably, the heating mechanism further includes a fixed base, a rocker arm, a connecting rod, a power source, and a connecting member that can be driven by the power source to move in a left-right direction. The fixed base is disposed on the base, the rocker arm is rotatably connected to the fixed base via a first rotating shaft, the power source is disposed on the rocker arm, the connecting member is rotatably connected to one end of the connecting rod via a second rotating shaft, the other end of the connecting rod is rotatably connected to the fixed base via a third rotating shaft, and the heating assembly is disposed on the connecting rod.

[0022] Furthermore, the power source includes a linear motor, and the connector is disposed on the output shaft of the linear motor; and / or, the center lines of the first rotating shaft, the second rotating shaft, and the third rotating shaft extend in the left-right direction and are parallel to each other but do not coincide; and / or, the heating mechanism further includes a support plate, the support plate being rotatably connected to the fixed base via a fourth rotating shaft, the center lines of the fourth rotating shaft and the third rotating shaft coinciding with each other, and the heating component being disposed on the support plate and the connecting rod.

[0023] According to a preferred embodiment, the nucleic acid extraction and amplification device further includes a magnet assembly, which includes a mounting base disposed on a base, a permanent magnet movably disposed on the mounting base, and an electromagnet for driving the permanent magnet to move, wherein the electromagnet is disposed on or inside the mounting base.

[0024] The present invention adopts the above solution, which has the following advantages compared with the prior art:

[0025] The nucleic acid extraction and amplification device of the present invention has a first driving mechanism and a second driving mechanism that can engage with two rotating pistons of a microfluidic chip. By controlling the chip mounting mechanism to move between the first and second driving mechanisms, the device can select whether to engage the first or second driving mechanism to connect specific chambers, so that the nucleic acid extraction reagent and the amplification reagent react according to the time sequence and sample mixing. The device can automatically perform nucleic acid extraction and amplification after the microfluidic chip is loaded, without the need for other manual operations. It is quick and convenient, and can effectively avoid contamination and human error. Attached Figure Description

[0026] To more clearly illustrate the technical solution of the present invention, the accompanying drawings used in the description of the embodiments will be briefly introduced below. Obviously, the accompanying drawings described below are only some embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0027] Figure 1 This is a three-dimensional structural diagram of a nucleic acid extraction and amplification device according to an embodiment of the present invention, wherein the cover is not shown;

[0028] Figure 2 for Figure 1 Front view of the nucleic acid extraction and amplification device shown;

[0029] Figure 3 for Figure 1 Top view of the nucleic acid extraction and amplification device shown;

[0030] Figure 4a and Figure 4b These are schematic diagrams of the microfluidic chip from two different perspectives.

[0031] Figure 5 This is a schematic diagram of the moving mechanism;

[0032] Figure 6 A three-dimensional structural diagram of the chip mounting mechanism, the third drive mechanism, and the moving mechanism;

[0033] Figure 7 for Figure 6 Side view of the mechanism shown;

[0034] Figure 8 A three-dimensional diagram of the chip mounting mechanism and the third drive mechanism;

[0035] Figure 9 This is a side view of the first drive mechanism;

[0036] Figure 10 for Figure 9 Sectional view along the middle AA direction;

[0037] Figure 11 This is a three-dimensional structural diagram of the heating mechanism;

[0038] Figure 12 Front view of the heating mechanism;

[0039] Figure 13 This is a side view of the heating mechanism;

[0040] Figure 14 This is a top view of the heating mechanism;

[0041] Figure 15 and Figure 16These are schematic diagrams of the optical inspection mechanism from two different perspectives.

[0042] Figure 17 This is a three-dimensional structural diagram of the magnet assembly;

[0043] Figure 18 This is a top view of the magnet assembly.

[0044] in,

[0045] 1. Base; 10. Base plate; 11. Mounting plate;

[0046] 2. Chip mounting mechanism; 20. Chip housing; 200. Chip slot; 201. Bottom wall; 202. Front wall; 203. Left wall; 2031. Folded edge; 2032. Through hole; 2033. Notch; 204. Right wall; 2041. Folded edge; 2042. Through hole; 2043. Notch; 205. Cutout area;

[0047] 3. First drive mechanism; 31. First drive pin; 32. Motor; 33. Worm gear transmission mechanism; 34. Gearbox;

[0048] 4. Second drive mechanism; 41. Second drive pin;

[0049] 5. Heating mechanism; 50. Heating component; 500. Radiator; 502. Heat-conducting plate; 503. Cooling fan; 504. Insulation cotton; 51. Mounting base; 52. Rocker arm; 53. Connecting rod; 54. Linear motor; 55. Connector; 56. Support plate; a. First rotating shaft; b. Second rotating shaft; c. Third rotating shaft; d. Fourth rotating shaft;

[0050] 6. Fluorescence detection mechanism; 60. Detection unit; 61. Horizontal guide rail; 62. Motor;

[0051] 7. Moving mechanism; 70. Horizontal guide rail; 71. Slide; 72. Vertical guide rail; 73. First chip motor; 74. Second chip motor; 740. Lead screw;

[0052] 8. Third drive mechanism; 80. Translation drive component; 801. Slot; 81. Guide rail; 82. Motor; 820. Lead screw;

[0053] 9. Magnet assembly: 90. Mounting base; 91. Mounting shaft; 92. Permanent magnet; 93. Sensor chip; 94. Proximity switch;

[0054] 100. Microfluidic chip; 101. Left rotating piston; 102. Right rotating piston; 103. Translation piston; 104. Separation region; 105. Lower part; 106. Step surface. Detailed Implementation

[0055] The preferred embodiments of the present invention will now be described in detail with reference to the accompanying drawings, so that the advantages and features of the present invention can be more readily understood by those skilled in the art. It should be noted that the description of these embodiments is for the purpose of aiding understanding the present invention, but does not constitute a limitation thereof.

[0056] In the description of this application, it should be understood that the terms "upper," "lower," "left," "right," "front," "rear," "vertical," "horizontal," "inner," and "outer," etc., indicating orientation or positional relationships based on the orientation or positional relationships shown in the accompanying drawings, are used 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, or be constructed and operated in a specific orientation, and therefore should not be construed as a limitation of this application. Figure 2 For example, left and right correspond to the left and right sides of the paper in the diagram, respectively; up and down correspond to the bottom and top sides of the paper in the diagram, respectively; and front and back correspond to directions perpendicular to the paper, with the side closer to the observer considered front. All directional terms used in this article are in the form of... Figure 2 The view shown is for reference only.

[0057] Figures 1 to 3 A nucleic acid extraction and amplification apparatus according to an embodiment of the present invention is shown. This apparatus is adapted to a microfluidic chip for nucleic acid extraction, purification, and PCR amplification. Figures 1 to 3 As shown, the nucleic acid extraction and amplification device includes a base 1 and a chip mounting mechanism 2, a first driving mechanism 3, a second driving mechanism 4, a heating mechanism 5, a moving mechanism 7, a third driving mechanism 8, and a magnet assembly 9, all mounted on the base 1. The device also enables fluorescence detection of the amplified products, and accordingly includes a fluorescence detection mechanism 6. Furthermore, the device includes a cover (not shown) mounted on the base 1 to enclose the aforementioned mechanisms and components. A slit-shaped through-hole, located directly above the chip mounting mechanism 2, is provided on the upper side wall of the cover to allow the microfluidic chip to pass through.

[0058] The structure of the microfluidic chip 100 is as follows Figure 4a and Figure 4bAs shown, the microfluidic chip 100 has multiple chambers, with microchannels between them for liquid to pass through. The microfluidic chip 100 includes a left rotating piston 101 and a right rotating piston 102, used to switch the connectivity of the microchannels. By rotating the left rotating piston 101 or the right rotating piston 102, some microchannels can be connected while others are disconnected. The microfluidic chip 100 also includes a translational piston 103, used to provide the driving force for liquid flow. As the translational piston 103 moves, negative or positive pressure can be applied to the liquid, thereby driving it to flow into a designated chamber. In this embodiment, the microfluidic chip 100 is a vertical microfluidic chip 100, with its left-right dimension (length) and its up-down dimension (height) being greater than its front-back dimension (width). The left rotating piston 101 is higher than the right rotating piston 102. Both are rotatably inserted into the body of the microfluidic chip 100 around their own axes. The left end of the left rotating piston 101 is exposed to facilitate engagement with the first drive mechanism 3, which is mainly used to control the switching between the chambers storing nucleic acid extraction and purification reagents and the purification separation chamber. The right end of the right rotating piston 102 is exposed to facilitate engagement with the second drive mechanism 4, which is mainly used to control the switching between the amplification reaction chamber and the amplification detection chamber. The translational piston 103 is movably inserted into the body of the microfluidic chip 100 in the left-right direction, with its left end extending beyond the body of the microfluidic chip 100 to facilitate engagement with the third drive mechanism 8. The upper part of the microfluidic chip 100 is provided with a purification separation chamber containing magnetic beads; correspondingly, the left side 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 part 105 of the microfluidic chip 100 is provided with a plurality of amplification detection cavities arranged side by side in the left-right direction. The portion of the microfluidic chip 100 body corresponding to the amplification detection cavity is transparent or semi-transparent to allow light to enter and excitation light to be emitted. The thickness of the lower part 105 of the microfluidic chip 100 is less than that of the upper part, and it has a step surface 106 facing downward.

[0059] The chip mounting mechanism 2 is used to mount the microfluidic chip 100, and it is movable on the base 1 in the left-right direction. Combined with... Figure 3 , Figure 5 and Figure 6As shown, the chip mounting mechanism 2 is mounted on the base 1 via a moving mechanism 7, and can move relative to the base 1 in both the left-right and up-down directions. The moving mechanism 7 includes a slide 71 movably connected to the base 1 in the left-right direction, and the chip mounting mechanism 2 is movably mounted on the slide 71 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 mounted 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 mounted 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. The slide 71 is connected to the output shaft of the first chip motor 73 via a lead screw. The moving mechanism 7 also includes a second chip motor 74 for driving the chip mounting mechanism 2 to move in the vertical direction. The chip mounting mechanism 2 is connected to the output shaft of the second chip motor 74 via a lead screw 740. The lead screw 740 extends in the vertical direction and is rotatably mounted on the mounting plate 11 around its own axis.

[0060] Combination Figures 6 to 8As shown, the chip mounting mechanism 2 includes a chip housing 20, within which a chip groove 200 for accommodating a microfluidic chip 100 is formed. The chip housing 20 includes a bottom wall 201, a front wall 202, a left wall 203, and a right wall 204, which together form the chip groove 200 for insertion of the microfluidic chip 100. The rear side of the left wall 203 has a right-bending flange 2031, and the rear side of the right wall 204 has a left-bending flange 2041. The flanges 2031 and 2041 serve to limit the movement of the microfluidic chip 100, preventing it from falling out from the rear side of the chip groove 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, thus forming a hollow portion 205 through which the lower part of the microfluidic chip 100 can 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. After the microfluidic chip 100 is installed in the chip housing 20, the stepped surface 106 abuts against the bottom wall 201. The lower part of the microfluidic chip 100 protrudes from the chip housing 20 and is located below the chip housing 20, which facilitates the fluorescence detection mechanism 6 to irradiate the amplification detection cavity at the bottom of the microfluidic chip 100 and collect fluorescence. The left end of the left rotating piston 101 of the microfluidic chip 100 is directly opposite 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 rotating piston 102 is directly opposite the through hole 2042 on the right wall 204, and the separation area 104 is directly opposite the notch 2033 on the left wall 203. The area to be heated of the microfluidic chip 100 faces the rear and is not blocked by the chip housing 20, so it can directly contact the heating mechanism 5.

[0061] like Figures 1 to 3 , Figure 9 , Figure 10 As shown, the first driving mechanism 3 is located on the left side of the chip mounting mechanism 2. The first driving mechanism 3 includes a first driving pin 31 for engaging with the left rotating piston 101 of the microfluidic chip 100 to drive its rotation. The second driving mechanism 4 is located on the right side of the chip mounting mechanism 2. The second driving mechanism 4 includes a second driving pin 41 for engaging with the right rotating piston 102 of the microfluidic chip 100 to drive its rotation. The chip mounting mechanism 2 has at least a first position for engaging the microfluidic chip 100 with the first driving pin 31 and disengaging it from the second driving pin 41, and a second position for engaging the microfluidic chip 100 with the second driving pin 41 and disengaging it from the first driving pin 31.

[0062] From a top-down view, the first drive mechanism 3 and the second drive mechanism 4 are centrally symmetrical. (The following is a summary...) Figure 9 and Figure 10The first drive mechanism 3 is described in detail; the second drive mechanism 4 is similar to the first drive mechanism 3. (Refer to...) Figure 9 and Figure 10 The first drive mechanism 3 includes a motor 32 mounted on the base 1 and a worm gear transmission mechanism 33 connecting the motor 32 and the first drive pin 31. The worm gear transmission mechanism 33 is housed within a reduction gearbox 34, which is fixedly mounted on the base plate 10 of the base 1. Specifically, the output shaft of the motor 32 extends in the front-rear direction, and the axis of the first drive pin 31 extends in the left-right direction. After the motor 32 operates, its output torque is redirected by the worm gear transmission mechanism 33, driving the first drive pin 31 to rotate, which in turn drives the left-side rotating piston 101 of the microfluidic chip 100 to rotate. The second drive mechanism 4 also includes a motor and a worm gear transmission mechanism, and its operating principle is similar to that of the first drive mechanism 3, so it will not be described in detail here.

[0063] The heating mechanism 5 is located behind the chip mounting mechanism 2 to heat the microfluidic chip 100. (Refer to...) Figures 11 to 14 As shown, the heating mechanism 5 includes a heating component 50 movably disposed on the base 1 and capable of approaching or moving 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, thereby being able to conform to the different heating areas.

[0064] The heating component 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 also includes a fixed base 51, a rocker arm 52, a connecting rod 53, a power source, and a connecting member 55 that can be driven by the power source to move in the left-right direction. The fixed base 51 is mounted on the base 1. The rocker arm 52 is rotatably connected to the fixed base 51 via a first rotating shaft a. The power source is mounted on the rocker arm 52. The connecting member 55 is rotatably connected to one end of the connecting rod 53 via a second rotating shaft b. The other end of the connecting rod 53 is rotatably connected to the fixed base 51 via a third rotating shaft c. The heating component 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 axes 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 coincide. The heating mechanism 5 also includes a support plate 56, which is rotatably connected to the fixed base 51 via a fourth rotating shaft d. The axis of the fourth rotating shaft d and the axis of the third rotating shaft c coincide. The heating component 50 is mounted on the support plate 56 and the connecting rod 53. The heating component 50 can swing in this way, allowing it to adhere to or detach from the microfluidic chip 100 without occupying too much space, resulting in a compact structure. Furthermore, the movable connection via the connecting rod 53 and other components avoids excessive frictional wear between parts.

[0065] The heating assembly 50 specifically includes a heat sink 500, multiple heating elements (not shown in the figure) disposed on the front side of the heat sink 500, heat-conducting plates 502 respectively covering each heating element, and a cooling fan 503 disposed on the rear side of the heat sink 500. The multiple heating elements are spaced apart vertically, and correspondingly, the multiple heat-conducting plates 502 are also spaced apart vertically. A first heating area is formed on one heat-conducting plate 502, and a second heating area is formed on the other heat-conducting plate 502 below it. The heating element is specifically a TEC cooling chip, embedded directly behind the heat-conducting plate 502. The heating assembly 50 also includes insulation cotton surrounding the heating elements. Each heating element can have a different temperature. By moving the chip mounting mechanism 2 up and down, the amplification reaction can be cycled at different temperatures according to the set reaction program.

[0066] The fluorescence detection mechanism 6 is located below the chip mounting mechanism 2, and is used to attach to or approach the lower part 105 of the microfluidic chip 100 to irradiate its amplification detection cavity and collect excitation light. (Refer to...) Figure 15 and Figure 16 As shown, the fluorescence detection mechanism 6 is movably mounted on the base 1 in the left-right direction. Specifically, the base plate 10 of the base 1 is provided with a horizontal guide rail 61 extending in the left-right direction. The fluorescence detection mechanism 6 is slidably mounted on the horizontal guide rail 61. The base 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 through a lead screw. By moving the fluorescence detection mechanism 6, the detection of all amplification detection cavities of the microfluidic chip 100 can be completed by one fluorescence detection mechanism 6. For example, in a specific example, the fluorescence detection mechanism 6 has four detection units 60 corresponding to four different colors of excitation light. Multiple detection units 60 are arranged side by side in the left-right direction. The microfluidic chip 100 has 12 amplification detection cavities arranged side by side 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 moved again to align with amplification detection cavities 9 to 12, thereby completing the detection of all amplification detection cavities.

[0067] Combination Figures 6 to 8As shown, the third driving mechanism 8 is disposed on the chip mounting mechanism 2. The third driving mechanism 8 includes a translational driving member 80 for engaging with the translational piston 103 of the microfluidic chip 100 to drive its movement. The translational driving member 80 is movably disposed on the chip mounting mechanism 2 in the left-right direction and is located on the right side of the chip mounting mechanism 2. The translational driving member 80 can move left and right together with the chip mounting mechanism 2, and can also move left and right relative to the chip mounting mechanism 2 to drive the translational piston 103 to slide left and right within the body of the microfluidic chip 100. The translation drive 80 has a slot 801 for inserting the translation piston 103, the slot 801 having an upward-facing opening; in its initial position, the translation drive 80 is embedded in the notch 2043 of the right wall 204 of the chip housing 20; when the microfluidic chip 100 is inserted into the chip housing 20, the right end of its translation piston 103 falls from top to bottom into the slot 801 of the translation drive 80 and engages with the translation drive 80; and throughout the detection process, the translation drive 80 and the translation piston 103 remain engaged with each other.

[0068] The third drive mechanism 8 also includes a guide rail 81 and a motor 82. The guide rail 81 extends in the left-right direction and is mounted on the chip mounting mechanism 2, specifically fixedly connected to the chip housing 20. The translation drive component 80 is slidably mounted on the guide rail 81. The motor 82 is mounted on the chip mounting mechanism 2 and is used to drive the translation drive component 80 to slide left and right; specifically, the motor is connected to the translation drive component 80 via a lead screw 820.

[0069] The magnet assembly 9 is used to apply a magnetic field to a magnetic bead within the purification and separation chamber of the microfluidic chip 100. With the left-side rotating piston 101 and the first drive pin 31 of the microfluidic chip 100 engaged, 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 within the magnetic field of the magnet assembly 9; in the second state, the purification and separation chamber is removed from the magnetic field of the magnet assembly 9. (Refer to...) Figure 17 and Figure 18As shown, the magnet assembly 9 includes a mounting base 90 disposed on the base 1, a permanent magnet 92 movably disposed on the mounting base 90, and an electromagnet (not shown in the figure) for driving the permanent magnet 92 to move. The electromagnet is disposed on or inside the mounting base 90. Specifically, the mounting base 90 is fixedly mounted on the reduction gearbox 34 of the first drive mechanism 3. The permanent magnet 92 is movably mounted on the mounting base 90 via a mounting shaft 91. The mounting shaft 91 extends in the left-right direction and is movably connected to the mounting base 90 in the left-right direction. The permanent magnet 92 is fixedly mounted on the left end of the mounting shaft 91 and faces the right side. An electromagnet is disposed in the mounting base 90 to drive the mounting shaft 91 to move. When the electromagnet is energized, the mounting shaft 91 extends to the right, driving 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 separation chamber. The magnet assembly 9 is in the first state. When the electromagnet is de-energized, the mounting shaft 91 retracts to the right, the permanent magnet 92 disengages from the separation area 104 of the microfluidic chip 100, and its magnetic field leaves the purification separation chamber. The magnet assembly 9 is in the second state. The mounting shaft 91 is also equipped with a sensor piece 93, and the mounting base 90 is equipped with a proximity switch 94 for detecting the position of the permanent magnet 92. The proximity switch 94 is specifically a photoelectric sensor. When the sensor piece 93 enters or leaves the detection area of ​​the photoelectric sensor, the photoelectric sensor is triggered.

[0070] The working process of this nucleic acid extraction and amplification device is as follows:

[0071] The microfluidic chip 100 is inserted into the chip slot 200. After insertion, the right end of the translation piston 103 falls into the slot 801 of the translation drive 80, so that the two engage.

[0072] When it is necessary to rotate the left rotary piston 101 to connect certain chambers, the chip mounting mechanism 2 moves to the left as a whole, so that the first drive pin 31 engages with the left end of the left rotary piston 101 (specifically, the first drive pin 31 passes through the through hole 2032 on the chip housing 20 and is inserted into the cross groove on the left end of the left rotary piston 101). The motor of the first drive mechanism 3 runs, and the first drive pin 31 rotates, thereby driving the left rotary piston 101 to rotate. At this time, the right rotary piston 102 disengages from the microfluidic chip 100. At the same time, the translation drive 80 and the translation piston 103 engage with each other. After the left rotary piston 101 connects certain chambers, the motor 82 of the third drive mechanism 8 runs, and the translation drive 80 moves left and right, thereby driving the translation piston 103 to move left and right within the microfluidic chip 100, thereby providing negative or positive pressure to push the liquid to flow between the connected chambers.

[0073] When it is necessary to rotate the right rotary piston 102 to connect other chambers, the chip mounting mechanism 2 moves to the right as a whole, so that the second drive pin 41 engages with the right end of the right rotary piston 102 (specifically, the second drive pin 41 passes through the through hole 2032 on the chip housing 20 and is inserted into the cross groove on the right end of the right rotary piston 102). The motor of the second drive mechanism 4 runs, and the second drive pin 41 rotates, thereby driving the right rotary piston 102 to rotate. At this time, the left rotary piston 101 and the permanent magnet 92 are both disengaged from the microfluidic chip 100. At the same time, the translation drive 80 and the translation piston 103 engage with each other. After the right rotary piston 102 connects some chambers, the motor 82 of the third drive mechanism 8 runs, and the translation drive 80 moves left and right, thereby driving the translation piston 103 to move left and right in the microfluidic chip 100, thereby providing negative or positive pressure to push the liquid to flow between the connected chambers.

[0074] When it is necessary to adsorb the magnetic beads (specifically located in a purification separation chamber) in the microfluidic chip 100 to separate the extracted nucleic acid (nucleic acid extraction), the chip mounting mechanism 2 is moved to the left to the designated position, the electromagnet of the magnet assembly 9 is energized, the mounting shaft 91 extends to the right, and 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 rotating piston 101 and the first driving pin 31 are engaged, and the translation piston 103 and the translation drive 80 are engaged. Rotating the left rotating piston 101 connects the sample extraction separation chamber and the waste liquid chamber, and then moving the translation piston 103 left and right allows the separated waste liquid to flow into the waste liquid chamber;

[0075] When the microfluidic chip 100 undergoes an amplification reaction, the linear motor 54 of the heating mechanism 5 operates, causing the heating component 50 to swing forward and adhere to the rear side of the microfluidic chip 100. After adhesion, according to the cycle program, the microfluidic chip 100 and the first heating area are adhered to each other for a period of time to react at a first temperature; the chip mounting mechanism 2 is moved downward to allow the microfluidic chip 100 and the second heating area to adhere to each other for a period of time to react at a second temperature; the chip mounting mechanism 2 is moved upward to allow the microfluidic chip 100 and the first heating area to adhere to each other, and this step is repeated according to the cycle number.

[0076] When fluorescence needs to be collected for detection, the chip mounting mechanism 2 is moved down, and the fluorescence detection mechanism 6 is aligned with the amplification detection chamber of the microfluidic chip 100. The fluorescence detection mechanism 6 is moved left and right to complete the detection of all amplification detection chambers.

[0077] The working principle of this nucleic acid extraction and amplification device for nucleic acid extraction and amplification is roughly as follows:

[0078] 1. Nucleic acid extraction and purification are performed through a control chip mounting mechanism, a first driving mechanism, a third driving mechanism, and a magnet assembly, as detailed below:

[0079] The microfluidic chip is inserted, and the chip mounting mechanism moves to the left, causing the left rotating piston to engage the first drive pin.

[0080] Drive the left rotating piston and translate the piston to introduce the lysis buffer into the purification and separation chamber, where it reacts with the sample.

[0081] After the reaction is complete, the magnet assembly applies a magnetic field to the purification and separation chamber. The lysed nucleic acids are adsorbed onto the magnetic beads, driving the left rotating piston and the translational piston, so that the unadsorbed cell debris and other debris are discharged into the waste liquid chamber along with the waste liquid.

[0082] The permanent magnet of the magnet assembly retracts, causing its magnetic field to leave the purification separation chamber; the left rotating piston and translational piston are driven to introduce the washing solution into the purification separation chamber, where the washing solution washes off the nucleic acid on the magnetic beads and performs rinsing.

[0083] After rinsing, the permanent magnet of the magnet assembly extends to apply a magnetic field to the purification and separation chamber. The rinsed nucleic acid is adsorbed onto the magnetic beads, driving the left rotating piston and the translational piston, so that the unadsorbed waste and other waste are discharged into the waste liquid chamber along with the waste liquid.

[0084] The above rinsing steps can be repeated multiple times;

[0085] The permanent magnet of the magnet assembly retracts, causing its magnetic field to leave the purification separation chamber; the left rotating piston and translational piston are driven to introduce the eluent into the purification separation chamber, where the eluent washes off the nucleic acids on the magnetic beads for subsequent amplification and detection.

[0086] 2. Nucleic acid amplification and detection are performed through a control chip mounting mechanism, a first driving mechanism, a second driving mechanism, a third driving mechanism, a heating mechanism, and a fluorescence detection mechanism, as detailed below:

[0087] Drive the left rotating piston and translate the piston to mix the amplification enzymes and other reagents with the separated nucleic acid;

[0088] The chip mounting mechanism moves to the right, causing the right-side rotating piston to engage the second drive pin.

[0089] Drive the right-side rotating piston and the translational piston to connect each amplification detection chamber in sequence, and distribute the nucleic acid and reaction solution into each amplification detection chamber.

[0090] The chip mounting mechanism is moved to the right to be directly opposite the heating mechanism;

[0091] Drive the heating mechanism to bring the heating component closer to the microfluidic chip;

[0092] The chip mounting mechanism moves up and down repeatedly, allowing the microfluidic chip to fit into two heating areas with different temperatures to meet the set PCR amplification cycle program.

[0093] The moving fluorescence detection mechanism allows it to sequentially collect fluorescence from all amplification detection chambers.

[0094] This nucleic acid extraction and amplification device can automatically perform nucleic acid extraction, purification, PCR amplification, and detection after the microfluidic chip 100 is loaded, without any other manual operation. The detection is quick and convenient, and it effectively avoids contamination and human error, resulting in relatively accurate test results. It is particularly suitable for nucleic acid extraction, purification, and detection. The various mechanisms of this nucleic acid extraction and amplification device are rationally arranged and compactly structured, making the device small in size and not occupying excessive space.

[0095] As indicated in this specification and claims, the terms "comprising" and "including" only indicate the inclusion of expressly identified steps and elements, and these steps and elements do not constitute an exclusive list; the 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.

[0096] It should be noted that, unless otherwise specified, when a feature is referred to as "fixed" or "connected" to another feature, it can be directly fixed or connected to the other feature, or indirectly fixed or connected to the other feature. Furthermore, the descriptions of "up," "down," "left," and "right" used in this invention are only relative to the relative positional relationships of the various components of the invention in the accompanying drawings.

[0097] It is further understood that the terms "first," "second," etc., are used to describe various types of information, but this information should not be limited to these terms. These terms are only used to distinguish information of the same type from one another, and do not indicate a specific order or degree of importance. In fact, the expressions "first," "second," etc., are completely interchangeable. For example, without departing from the scope of this disclosure, first information can also be referred to as second information, and similarly, second information can also be referred to as first information.

[0098] The above embodiments are merely illustrative of the technical concept and features of the present invention, and are preferred embodiments. Their purpose is to enable those skilled in the art to understand the content of the present invention and implement it accordingly, and they should not be construed as limiting the scope of protection of the present invention. All equivalent transformations or modifications made according to the principles of the present invention should be covered within the scope of protection of the present invention.

Claims

1. A nucleic acid extraction amplification device, characterized by, The nucleic acid extraction and amplification device comprises: a chip mounting mechanism for mounting a microfluidic chip, the chip mounting mechanism being capable of moving in left-right direction; a first driving mechanism located at the left side of the chip mounting mechanism, the first driving mechanism comprising a first driving pin for engaging with a rotating piston at the left side of the microfluidic chip to drive the rotating piston to rotate; and a second driving mechanism located at the right side of the chip mounting mechanism, the second driving mechanism comprising a second driving pin for engaging with a rotating piston at the right side of the microfluidic chip to drive the rotating piston to rotate; the chip mounting mechanism having at least a first position for engaging the microfluidic chip with the first driving pin and disengaging the microfluidic chip from the second driving pin, and a second position for engaging the microfluidic chip with the second driving pin and disengaging the microfluidic chip from the first driving pin; the nucleic acid extraction and amplification device further comprising a heating mechanism for heating the microfluidic chip, the heating mechanism comprising a heating assembly having a first heating area and a second heating area located below the first heating area, the first heating area and the second heating area having different temperatures, the chip mounting mechanism having a position capable of abutting the first heating area and a position capable of abutting the second heating area; the nucleic acid extraction and amplification device further comprising a third driving mechanism, the third driving mechanism comprising a translation driving member for engaging with a translation piston of the microfluidic chip to drive the translation piston to move, the translation driving member being movably arranged on the chip mounting mechanism in left-right direction, the translation driving member being located at the left side or the right side of the chip mounting mechanism; the nucleic acid extraction and amplification device further comprising a magnet assembly for applying a magnetic field to a purification separation chamber in the microfluidic chip; the chip mounting mechanism comprising a chip shell, the chip shell comprising a bottom wall, a front wall, a left wall and a right wall, the four walls forming a chip slot for inserting the microfluidic chip, the rear side of the left wall having a bent edge extending to the right, the rear side of the right wall having a bent edge extending to the left, the rear edge of the bottom wall being located at a distance in front of the bent edges of the left wall and the right wall, thereby forming a hollow part for the lower part of the microfluidic chip to pass through, the left wall being provided with a through hole for the first driving pin to pass through, the right wall being provided with a through hole for the second driving pin to pass through; the left wall or the right wall of the chip shell being provided with a notch for cooperating with the translation driving member, the translation driving member having an initial position embedded in the notch; the heating area of the microfluidic chip being configured to face the rear side and not being shielded by the chip shell, so as to be capable of directly contacting the heating mechanism.

2. The nucleic acid extraction amplification device according to claim 1, characterized by The first driving mechanism or the second driving mechanism comprises a motor and a worm gear transmission mechanism for connecting the motor and the first driving pin or the second driving pin.

3. The nucleic acid extraction amplification device according to claim 2, characterized by The first driving mechanism and the second driving mechanism are center-symmetrical in the top view.

4. The nucleic acid extraction amplification device according to claim 1, characterized by The chip mounting mechanism is movably arranged on a base along left-right direction by a moving mechanism, the base is provided with a horizontal guide rail extending along left-right direction, and the moving mechanism comprises a sliding seat movably arranged on the horizontal guide rail along left-right direction.

5. The nucleic acid extraction amplification device according to claim 4, wherein The moving mechanism further comprises a vertical guide rail arranged on the sliding seat, and the chip mounting mechanism is movably arranged on the vertical guide rail along up-down direction.

6. The nucleic acid extraction amplification device according to claim 1, wherein The translation driving part has a clamping groove for inserting the translation piston, and the clamping groove has a slot opening facing upward; the third driving mechanism comprises a guide rail extending along left-right direction arranged on the chip mounting mechanism and a motor for driving the translation driving part to move, and the translation driving part is slidably arranged on the guide rail.

Images