CN112522085A

CN112522085A - Gene detection device based on solid-state nanopore

Info

Publication number: CN112522085A
Application number: CN202011391311.7A
Authority: CN
Inventors: 龚秀清
Original assignee: Individual
Current assignee: Hefei Guoyan Hanyin Testing Technology Co ltd
Priority date: 2020-12-01
Filing date: 2020-12-01
Publication date: 2021-03-19
Anticipated expiration: 2040-12-01
Also published as: CN112522085B

Abstract

The invention discloses a gene detection device based on solid-state nanopores, which belongs to the technical field of gene detection equipment and comprises a top plate, an installation box, two adjusting boxes, a nanopore chip and a workbench, wherein the adjusting boxes are arranged between the top plate and the installation box, the top plate and the installation box are fixedly connected, the workbench is arranged in the middle of the upper surface of the installation box, the two adjusting boxes are symmetrically arranged, threaded upright columns are movably arranged in the adjusting boxes, threaded lifting blocks are connected to the surfaces of the threaded upright columns in a threaded mode, and connecting blocks are fixedly connected to one sides of the threaded lifting blocks. This gene detection device based on solid-state nanopore carries out the centre gripping through setting up the detection clamp to the nanopore chip and fixes to use the screw thread stand to accomplish detection achievement in the middle of inserting the nanopore chip into the liquid bath, the location accuracy and the reliability of nanopore chip when having improved the clamping, this has important meaning to reducing the experimental time, improving research efficiency.

Description

Gene detection device based on solid-state nanopore

Technical Field

The invention belongs to the technical field of gene detection, and particularly relates to a gene detection device based on solid-state nanopores.

Background

With the rise of the fourth generation sequencing technology, the research of the nanopore detection technology has a profound influence on the development of the biomolecule sensing and detection field. The improvement of experimental instruments and devices and the simplification of experimental steps have very important meanings for improving experimental efficiency and accelerating research process.

In the prior art, a device for clamping a nanopore chip in a laboratory is mainly composed of two liquid pools which are different in shape, one convex and one concave to realize positioning in one moving dimension, a rubber gasket for sealing, an external bolt frame for clamping the liquid pools, and a tightening bolt. The clamping steps mainly comprise: 1. placing the chip on a rubber gasket stuck on the liquid pool by using tweezers and positioning the chip by visual inspection; 2. another liquid pool adhered with a gasket is buckled on the chip, and is positioned by visual observation and is pre-clamped by hands; 3. the two liquid pools are placed on an external bolt frame, and the bolts are screwed down to axially move and tightly press the two liquid pools. But the method is extremely easy to cause pollution and even damage to the chip and has irreparable influence on the experiment. The operation requirement on personnel is high, the experimental efficiency is greatly influenced, and the research process is slowed down.

For this reason, a solid-state nanopore-based gene detection device needs to be proposed to solve the above problems.

Disclosure of Invention

In view of the deficiencies of the prior art, the present invention provides a solid-state nanopore-based gene detection device to solve the problems set forth in the background art.

In order to achieve the purpose, the invention provides the following technical scheme: a gene detection device based on solid-state nanopores comprises a top plate, an installation box, an adjusting box, a nanopore chip and a workbench, wherein the adjusting box is arranged between the top plate and the installation box, the top plate and the installation box are fixedly connected, the workbench is arranged in the middle of the upper surface of the installation box, the two adjusting boxes are symmetrically arranged, the inner part of each adjusting box is movably provided with a threaded upright post, the surface of the threaded upright post is in threaded connection with a threaded lifting block, one side of the threaded lifting block is fixedly connected with a connecting block, a connecting plate is fixedly arranged between the connecting blocks, an equipment plate is fixedly arranged on the lower surface of the connecting plate, the equipment board is provided with a detection clamp for clamping a nanopore chip, a left liquid pool and a right liquid pool which are positioned on the workbench are arranged below the detection clamp, and the nanopore chip is inserted in the middle of the left liquid pool and the right liquid pool.

This technical scheme is further optimized, the top swing joint of screw thread stand is at the lower surface of roof, the bottom swing joint of screw thread stand is in the inside of install bin.

Further optimize this technical scheme, the inside fixed mounting of install bin has the motor case, the equal fixed mounting in both sides of motor case has driving motor for provide drive power.

Further optimize this technical scheme, the regulating tank has been seted up on the surface of regulating box inboard, the screw thread elevator exposes on the surface of regulating tank, the connecting block passes the regulating tank and is connected fixedly with the screw thread elevator.

Further optimizing the technical scheme, the clamping end of the detection clamp is sleeved with a protective sleeve, and the protective sleeve is used for protecting the nanopore chip from being damaged.

Further optimize this technical scheme, be provided with the cushion collar that is used for the buffering between the inboard of detecting two arms of clamp, the top winding that detects the clamp has about band, the other end winding of about band has the jack, jack fixed mounting is on the equipment board.

Further optimize this technical scheme, the surface of equipment board has been seted up and has been detected the clamp spout, the inside sliding connection that detects the clamp spout has the connection ball, one side fixed connection that detects the clamp is on the surface of connecting the ball.

Further optimize this technical scheme, driving motor's output shaft fixedly connected with action wheel, the meshing has from the driving wheel on the action wheel, from carrying out fixed connection with the screw thread pillar from the driving wheel.

Further optimizing the technical scheme, the nanopore chip is a chip with a solid nanopore, and the chip with the solid nanopore is placed on the chip clamp.

Compared with the prior art, the invention provides a gene detection device based on solid-state nanopores, which has the following beneficial effects:

this gene detection device based on solid-state nanopore carries out the centre gripping through setting up the detection clamp to the nanopore chip and fixes to use the screw thread stand to accomplish detection achievement in the middle of inserting the nanopore chip into the liquid bath, the location accuracy and the reliability of nanopore chip when having improved the clamping, this has important meaning to reducing the experimental time, improving research efficiency.

In addition, by adopting the gene detection device, the work of inserting the nanopore chip into the liquid pool is completed by the detection clamp, the nanopore chip is clamped and fixed by the detection clamp, and the nanopore chip is inserted into the middle of the liquid pool by the threaded upright post to complete the detection work, so that the steps of manual operation are reduced, the pollution source in the detection is reduced, and the detection accuracy is improved.

Drawings

FIG. 1 is a schematic structural diagram of a solid-state nanopore-based genetic testing device according to the present invention;

FIG. 2 is a schematic view of a connecting structure of a detecting clip chute of the gene detecting device based on solid-state nanopores according to the present invention;

FIG. 3 is a schematic diagram of a nanopore chip of the solid-state nanopore-based genetic testing device according to the present invention;

FIG. 4 is a schematic view of the connection structure of the detection clip of the gene detection device based on solid-state nanopore according to the present invention;

FIG. 5 is a schematic structural diagram of a regulation groove of the solid-state nanopore-based gene detection device according to the present invention;

FIG. 6 is a schematic structural diagram of a driving wheel and a driven wheel of the gene detection device based on solid-state nanopore according to the present invention;

FIG. 7 is a record of the current drop event and its sequence information observed after the DNA molecule is captured by the nanopore in example three.

In the figure: 1. a top plate; 2. installing a box; 3. an adjusting box; 4. a work table; 5. a motor case; 6. a drive motor; 7. a driving wheel; 8. a driven wheel; 9. a threaded upright post; 10. a threaded lifting block; 11. connecting blocks; 12. a connecting plate; 13. an equipment board; 14. detecting a clamp; 15. a protective sleeve; 16. a buffer sleeve; 17. a retraction mechanism; 18. a restraint band; 19. a nanopore chip; 20. a left liquid pool; 21. a right liquid tank; 22. detecting a clamp chute; 23. a connecting ball; 24. and (4) adjusting the groove.

Detailed Description

In order to make the technical solutions better understood by those skilled in the art, the technical solutions in the embodiments of the present application will be clearly and completely described below with reference to the drawings in the embodiments of the present application, and it is obvious that the described embodiments are only partial embodiments of the present application, but not all embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present application.

It should be noted that the terms "first," "second," and the like in the description and claims of this application and in the drawings described above are used for distinguishing between similar elements and not necessarily for describing a particular sequential or chronological order. It should be understood that the data so used may be interchanged under appropriate circumstances such that embodiments of the application described herein may be used. Furthermore, the terms "comprises," "comprising," and "having," and any variations thereof, are intended to cover a non-exclusive inclusion, such that a process, method, system, article, or apparatus that comprises a list of steps or elements is not necessarily limited to those steps or elements expressly listed, but may include other steps or elements not expressly listed or inherent to such process, method, article, or apparatus.

In this application, the terms "upper", "lower", "left", "right", "front", "rear", "top", "bottom", "inner", "outer", "middle", "vertical", "horizontal", "lateral", "longitudinal", and the like indicate orientations or positional relationships based on the orientations or positional relationships shown in the drawings. These terms are used primarily to better describe the present application and its embodiments, and are not used to limit the indicated devices, elements or components to a particular orientation or to be constructed and operated in a particular orientation.

Moreover, some of the above terms may be used to indicate other meanings besides the orientation or positional relationship, for example, the term "on" may also be used to indicate some kind of attachment or connection relationship in some cases. The specific meaning of these terms in this application will be understood by those of ordinary skill in the art as appropriate.

Furthermore, the terms "mounted," "disposed," "provided," "connected," and "sleeved" are to be construed broadly. For example, it may be a fixed connection, a removable connection, or a unitary construction; can be a mechanical connection, or an electrical connection; may be directly connected, or indirectly connected through intervening media, or may be in internal communication between two devices, elements or components. The specific meaning of the above terms in the present application can be understood by those of ordinary skill in the art as appropriate.

It should be noted that the embodiments and features of the embodiments in the present application may be combined with each other without conflict. The present application will be described in detail below with reference to the embodiments with reference to the attached drawings.

The first embodiment is as follows:

referring to fig. 1-6, a gene detection device based on solid-state nanopores comprises a top plate 1, an installation box 2, an adjustment box 3, a nanopore chip 19 and a workbench 4, wherein the adjustment box 3 is arranged between the top plate 1 and the installation box 2, the top plate 1 and the installation box 2 are fixedly connected, the workbench 4 is arranged in the middle of the upper surface of the installation box 2, the two adjustment boxes 3 are symmetrically arranged, a threaded upright post 9 is movably arranged in the adjustment box 3, a threaded lifting block 10 is in threaded connection with the surface of the threaded upright post 9, the threaded lifting block 10 is used for driving a detection clamp 14 to move up and down, so that the nanopore chip 19 is inserted and taken out, a connecting block 11 is fixedly connected to one side of the threaded lifting block 10, and a connecting plate 12 is fixedly installed between the connecting blocks 11, the lower fixed surface of connecting plate 12 installs equipment board 13, equipment board 13 is provided with the detection clamp 14 that is used for centre gripping nanopore chip 19, the below that detects clamp 14 is provided with left liquid pool 20 and right liquid pool 21 that are located on workstation 4, nanopore chip 19 pegs graft in the middle part of left liquid pool 20 and right liquid pool 21 to carry out nanopore chip 19's detection work.

Specifically, the top swing joint of screw thread stand 9 is at the lower surface of roof 1, the bottom swing joint of screw thread stand 9 has guaranteed that screw thread stand 9 can rotate to drive and detect that clamp 14 reciprocates in the inside of install bin 2.

Specifically, the inside fixed mounting of install bin 2 has motor case 5, the equal fixed mounting in both sides of motor case 5 has driving motor 6 for it provides drive power to rotate for threaded upright post 9.

Specifically, adjustment tank 24 has been seted up on the surface of adjusting box 3 inboard, screw thread elevator 10 exposes on the surface of adjustment tank 24, connecting block 11 passes adjustment tank 24 and is connected fixedly with screw thread elevator 10, conveniently detects clamp 14 and reciprocates and drives nanopore chip 19 and carry out the work that detects.

Specifically, the clamping end of the detection clamp 14 is sleeved with a protective sleeve 15, the protective sleeve 15 is used for protecting the nanopore chip 19 from being damaged, and the positioning accuracy and reliability of the nanopore chip 19 during clamping are improved.

Specifically, a buffer sleeve 16 for buffering is arranged between the inner sides of two arms of the detection clamp 14, a restraint belt 18 is wound on the top end of the detection clamp 14, a retraction mechanism 17 is wound on the other end of the restraint belt 18, the retraction mechanism 17 is fixedly mounted on the equipment board 13, when the retraction mechanism 17 retracts the restraint belt 18, the restraint belt 18 drives the detection clamp to fix the nanopore chip 19, and the nanopore chip 19 is inserted into the liquid pool to complete detection.

Specifically, the surface of the equipment board 13 is provided with a detection clamp sliding groove 22, the inside sliding connection of the detection clamp sliding groove 22 is provided with a connecting ball 23, one side fixed connection of the detection clamp 14 is arranged on the surface of the connecting ball 23, and the stability of the detection clamp 14 is ensured when the detection clamp 14 operates.

Specifically, an output shaft of the driving motor 6 is fixedly connected with a driving wheel 7, a driven wheel 8 is meshed with the driving wheel 7, and the driven wheel 8 is fixedly connected with a threaded support 9, so that the driving motor 6 can drive the threaded support 9 to rotate.

Specifically, the nanopore chip 19 is a chip with a solid nanopore, and the chip with the solid nanopore is placed on a chip holder, so that the purpose of gene detection by using the solid nanopore is achieved.

Example two:

a gene detection device based on solid-state nanopores comprises a top plate 1, an installation box 2, an adjusting box 3, nanopore chips 19 and a workbench 4, wherein the adjusting box 3 is arranged between the top plate 1 and the installation box 2, the top plate 1 and the installation box 2 are fixedly connected, the workbench 4 is arranged in the middle of the upper surface of the installation box 2, the adjusting box 3 is provided with two adjusting boxes 3, the two adjusting boxes 3 are symmetrically arranged, a threaded upright column 9 is movably arranged in the adjusting box 3, a threaded lifting block 10 is in threaded connection with the surface of the threaded upright column 9, the threaded lifting block 10 is used for driving a detection clamp 14 to move up and down, so that the nanopore chips 19 are inserted and taken out, a connecting block 11 is fixedly connected to one side of the threaded lifting block 10, a connecting plate 12 is fixedly installed between the connecting blocks 11, and a device plate 13 is fixedly installed on the lower surface of the connecting plate 12, the equipment board 13 is provided with a detection clamp 14 for clamping a nanopore chip 19, a left liquid pool 20 and a right liquid pool 21 which are positioned on the workbench 4 are arranged below the detection clamp 14, and the nanopore chip 19 is inserted in the middle of the left liquid pool 20 and the right liquid pool 21, so that the detection work of the nanopore chip 19 is performed.

Specifically, under the condition that the internal space of the installation box 2 allows, the output shaft of the driving motor 6 can be directly connected to the bottom end of the threaded strut 9 without being fixedly connected with the driving wheel 7, so as to drive the threaded strut 9 to rotate.

Example three: detailed experiments

Using the apparatus of example 1, 5kbp was used

And the samples were diluted to a concentration of 1nM in 1M KCl, 10mM Tris and 1mM EDTA, all solutions were filtered using 0.2 μ M to remove large particles. The ionic current was measured by applying a voltage bias across the two ends, one Ag/AgCl electrode placed at the nanopore capillary and the other at the reservoir in the main fluid channel. A5 kbp sample was injected into the nanopore chip at a flow rate of 1 μ L/min with a 500mV bias. The occurrence of a recorded drop in current can be clearly seen, indicating that the passage of DNA molecules through the vicinity of the nanopore causes a drop in current density. By adopting the device, the sequence information of the tested DNA molecules can be read, and compared with the conventional detection device, the detection time is shorter and the accuracy is higher.

The invention has the beneficial effects that: this gene detection device based on solid-state nanopore carries out the centre gripping through setting up detection clamp 14 to nanopore chip 19 and fixes to use screw thread stand 9 to accomplish detection achievement in the middle of inserting nanopore chip 19 into the liquid bath, nanopore chip 19's location accuracy and reliability when having improved the clamping have reduced manual operation's step, have reduced the source of polluting in the detection, have improved and have detected the accuracy, and this has important meaning to reducing the test time, improving research efficiency.

Although embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that changes, modifications, substitutions and alterations can be made in these embodiments without departing from the principles and spirit of the invention, the scope of which is defined in the appended claims and their equivalents.

Claims

1. The gene detection device based on the solid-state nanopore is characterized by comprising a top plate (1), an installation box (2), an adjusting box (3), a nanopore chip (19) and a workbench (4), wherein the adjusting box (3) is arranged between the top plate (1) and the installation box (2) and is used for fixedly connecting the top plate (1) and the installation box (2), the workbench (4) is arranged in the middle of the upper surface of the installation box (2), the adjusting box (3) is provided with two adjusting boxes (3), the two adjusting boxes (3) are symmetrically arranged, a threaded upright post (9) is movably arranged in the adjusting box (3), a threaded lifting block (10) is in threaded connection with the surface of the threaded upright post (9), a connecting block (11) is fixedly connected to one side of the threaded lifting block (10), and a connecting plate (12) is fixedly arranged between the connecting blocks (11), the lower fixed surface of connecting plate (12) installs equipment board (13), equipment board (13) are provided with detection clamp (14) that are used for centre gripping nanopore chip (19), the below that detects clamp (14) is provided with left liquid bath (20) and right liquid bath (21) that are located workstation (4), nanopore chip (19) are pegged graft in the middle part of left liquid bath (20) and right liquid bath (21).

2. The gene detection device based on the solid-state nanopore as claimed in claim 1, wherein the top end of the threaded column (9) is movably connected to the lower surface of the top plate (1), and the bottom end of the threaded column (9) is movably connected to the inside of the installation box (2).

3. The gene detecting device based on the solid-state nanopore as claimed in claim 1, wherein a motor box (5) is fixedly installed inside the installation box (2), and driving motors (6) are fixedly installed on both sides of the motor box (5) and used for providing driving power.

4. The gene detection device based on the solid-state nanopore as claimed in claim 1, wherein the surface of the inner side of the adjusting box (3) is provided with an adjusting groove (24), the threaded lifting block (10) is exposed on the surface of the adjusting groove (24), and the connecting block (11) passes through the adjusting groove (24) to be fixedly connected with the threaded lifting block (10).

5. The gene detection device based on solid-state nanopore as claimed in claim 1, wherein a protecting sleeve (15) is sleeved on the clamping end of the detection clamp (14), and the protecting sleeve (15) is used for protecting the nanopore chip (19) from being damaged.

6. The gene detection device based on the solid-state nanopore as claimed in claim 1, wherein a buffer sleeve (16) for buffering is arranged between the inner sides of the two arms of the detection clip (14), a restraining belt (18) is wound on the top end of the detection clip (14), a retraction mechanism (17) is wound on the other end of the restraining belt (18), and the retraction mechanism (17) is fixedly mounted on the device board (13).

7. The gene detection device based on the solid-state nanopore as claimed in claim 1, wherein a detection clip sliding groove (22) is formed on the surface of the device plate (13), a connection ball (23) is slidably connected inside the detection clip sliding groove (22), and one side of the detection clip (14) is fixedly connected to the surface of the connection ball (23).

8. The gene detection device based on the solid-state nanopore as claimed in claim 3, wherein an output shaft of the driving motor (6) is fixedly connected with a driving wheel (7), a driven wheel (8) is engaged on the driving wheel (7), and the driven wheel (8) is fixedly connected with a threaded pillar (9).

9. The solid-state nanopore based gene detection device of claim 1, wherein the nanopore chip (19) is a solid-state nanopore chip, and the solid-state nanopore chip is placed on a chip holder.