CN216946986U

CN216946986U - Multi-head integrated nucleic acid extraction test tube and extraction and detection device

Info

Publication number: CN216946986U
Application number: CN202121181883.2U
Authority: CN
Inventors: 王德明; 周杰锋
Original assignee: Ningbo Kangcheng Deno Biomedical Co ltd
Current assignee: Ningbo Kangcheng Deno Biomedical Co ltd
Priority date: 2021-05-28
Filing date: 2021-05-28
Publication date: 2022-07-12
Anticipated expiration: 2031-05-28

Abstract

The utility model relates to the field of nucleic acid extraction and detection, in particular to a multi-head integrated nucleic acid extraction test tube and an extraction and detection device. The test tube includes a main tube; the inner cavity of the main pipe is sequentially provided with a cracking area, a washing area and an elution area from top to bottom; hydrophobic sealing layers are respectively arranged between the cracking zone and the washing zone and between the washing zone and the elution zone for separation, and magnetic bead washing liquid and nucleic acid eluent are respectively arranged in the washing zone and the elution zone; the inner cavity of the main pipe is also provided with a magnetic track extending from the cracking area to the elution area; a plurality of partition plates are arranged in the inner cavity of the elution area to divide the elution area into a plurality of mutually independent branch pipes; this test tube improves nucleic acid extraction efficiency for it is more steady during the magnetic bead motion. Meanwhile, a plurality of independent branch pipes are arranged at the bottom end of the main pipe to elute the magnetic beads respectively, so that the accuracy of extraction results is improved.

Description

Multi-head integrated nucleic acid extraction test tube and extraction and detection device

Technical Field

The utility model relates to the field of nucleic acid extraction and detection, in particular to a multi-head integrated nucleic acid extraction test tube.

Background

With the popularization of gene detection, personalized drug delivery, prenatal diagnosis and the like, the limitation of the traditional DNA extraction method becomes more and more obvious today when high throughput and automation are pursued in various fields of the biological industry. Because the magnetic bead method for extracting nucleic acid can realize automatic extraction and large-scale operation, and has simple operation and short time, the magnetic bead method for extracting nucleic acid is more and more emphasized.

The Chinese invention patent application (publication No. CN108796038B, published: 20191018) discloses a nucleic acid integrated detection method and a detection reagent tube, wherein a plurality of separating plugs which are arranged up and down are arranged in a main tube, and a liquid-phase or solid-phase hydrophobic layer is arranged on each separating plug, so that lysate, cleaning liquid and reaction liquid in the detection reagent tube are isolated; adding a sample into a lysis solution for uniform mixing and lysis, extracting nucleic acid in the sample by using nano magnetic beads, driving the nano magnetic beads to carry the nucleic acid to sequentially pass through each hydrophobic layer along a magnetic bead channel on the inner wall of a detection reagent tube by using an external magnetic body to enter cleaning solution and reaction solution, and realizing the cleaning and amplification of the nucleic acid, wherein a biological reagent required by the reaction solution is stored in a separation plug above the reaction solution, and finally, external equipment realizes the detection of the sample nucleic acid through optical detection, thereby realizing multiple steps of nucleic acid extraction, cleaning and amplification reaction in the same detection reagent tube. The utility model has the characteristics of reducing detection errors and reducing operation difficulty.

The prior art has the following defects: the number of the magnetic bead channels on the inner wall of the main pipe is small, the width of the magnetic bead channels is narrow, and the number of the magnetic beads driven by the magnet from the outer wall of the main pipe at one time is small, so that the nucleic acid extraction efficiency is low; the magnetic bead channel width is narrower to make and pile up each other easily between the magnetic bead and can not evenly distributed at the magnetic bead channel inner wall to be unstable when causing the magnetic bead motion. Meanwhile, the magnetic bar drives all the washed magnetic beads to elute in the same elution area at the same time, and the magnetic beads cannot be separately eluted to compare results, so that error influence factors are eliminated, and the accuracy of extraction results is reduced.

Disclosure of Invention

The purpose of the utility model is: aiming at the problems, the magnetic track is arranged in the main pipe, and the magnetic rod is inserted into the inner hole of the magnetic track to drive the magnetic bead to be attached to the outer wall of the magnetic track and move along with the magnetic rod; therefore, the magnetic beads can move downwards without being limited by the number of magnetic bead channels, a large number of magnetic beads are driven at one time, and the nucleic acid extraction efficiency is improved; the magnetic beads are uniformly distributed on the outer wall of the magnetic track with larger area and are not stacked, so that the magnetic beads are more stable in movement. Simultaneously, set up a plurality of independent branch pipes respectively and elute the magnetic bead in being responsible for the bottom, improve a multihead integration nucleic acid extraction test tube of extraction result accuracy.

In order to achieve the purpose, the utility model adopts the following technical scheme:

a multi-headed integrated nucleic acid extraction cuvette comprising a main tube; the inner cavity of the main pipe is sequentially provided with a cracking area, a washing area and an elution area from top to bottom; hydrophobic sealing layers are respectively arranged between the cracking zone and the washing zone and between the washing zone and the elution zone for separation, and magnetic bead washing liquid and nucleic acid eluent are respectively arranged in the washing zone and the elution zone; the inner cavity of the main pipe is also provided with a magnetic track extending from the cracking zone to the elution zone, one end of the magnetic track is closed, and the other end of the magnetic track is provided with an opening for the magnetic rod to extend into; the inner cavity of the elution zone is provided with a plurality of partition plates, and the radial two ends of the plurality of partition plates are respectively connected with the inner wall of the cavity of the elution zone and the outer wall of the magnetic track, so that the elution zone is divided into a plurality of mutually independent branch pipes; two adjacent clapboards are distributed oppositely, the inner wall of the cavity of the elution area and the outer wall of the magnetic track are distributed oppositely to form four walls of the branch pipe, and the bottom surface of the elution area is connected with the bottom ends of the four walls of the branch pipe to form the bottom surface of the branch pipe.

In addition, the utility model also discloses an integrated nucleic acid extraction and detection device, which comprises the integrated nucleic acid extraction test tube and a magnetic rod, wherein the outer wall of the magnetic rod is matched with the inner hole of the magnetic track.

Preferably, the main pipe inner cavity is provided with a first separating plug and a second separating plug; annular gaps are formed among the outer wall of the magnetic track, the first separating plug and the second separating plug; the hydrophobic sealing layers are respectively positioned in the first separating plug and the second separating plug and fill and seal the inner cavity of the main pipe at the position.

Preferably, the hydrophobic sealing layer is a paraffin or liquid-phase hydrophobic layer; the first and second plugs are both of an elastomeric material. The bottom ends of the branch pipes are mutually independent cylindrical. The plurality of branch pipes are distributed along the center of the main pipe in a circumferential shape; the axial central axis of the magnetic track is axially coincident with the axial central axis of the main pipe.

Preferably, the first separating plug comprises a first plug body, a first arc surface, a first prism surface and a first central hole; the upper end of the inner wall of the main pipe is in an arc shape, and the lower end of the inner wall of the main pipe is in a hexagonal prism shape; the first arc surface is in an arc shape and is in interference fit with the arc inner wall at the upper end of the main pipe, and the first prismatic surface is in a hexagonal prism shape and is in interference fit with the hexagonal prism inner wall at the lower end of the main pipe; the first central hole is penetrated by the magnetic track and a gap between the first central hole and the outer wall of the magnetic track is filled and sealed by a hydrophobic sealing layer.

Preferably, the second septum plug comprises a second plug body, a second prismatic surface and a second central bore; the inner wall of the main pipe is also provided with a second step surface; the bottom surface of the second plug body in the horizontal direction is in contact with the second step surface in the horizontal direction; the second prismatic surface is in interference fit with the inner wall of the hexagonal prism at the lower end of the main pipe, the second central hole is penetrated by the magnetic track, and a gap between the second central hole and the outer wall of the magnetic track is filled and sealed by the hydrophobic sealing layer.

Preferably, the top end of the main pipe is provided with a pipe cover, and the pipe cover is screwed with the external thread of the outer wall of the main pipe through the internal thread; the pipe cover is provided with a positioning hole which vertically penetrates through, and the positioning hole is matched with the outer wall of the magnetic track.

The multi-head integrated nucleic acid extraction test tube adopting the technical scheme has the advantages that:

the hydrophobic sealing layer separates cell lysis solution in the lysis zone from magnetic bead washing solution in the washing zone and separates magnetic bead washing solution in the washing zone from nucleic acid eluent in the elution zone; after the magnetic rod is placed into the inner hole of the magnetic track from the top opening of the magnetic track, the magnetic beads in the cracking area are sucked and attached to the outer wall of the magnetic track by using the magnetic force of the magnetic rod; the magnetic bar moves downwards to drive the magnetic beads to move downwards to penetrate through the hydrophobic sealing layer to enter a washing area and an elution area respectively for washing and elution so as to complete the extraction of nucleic acid; in the mode, the magnetic beads are attached to the outer wall of the magnetic track and move along with the magnetic rod, the outer wall of the whole magnetic track can be used for moving the magnetic beads without being limited by the number of magnetic bead channels, a large number of magnetic beads can be driven at one time, and the nucleic acid extraction efficiency is improved; meanwhile, the magnetic beads are uniformly distributed on the outer wall of the magnetic track with larger area and are not stacked, so that the magnetic beads are more stable in movement. In addition, in the mode, the magnetic beads on the magnetic rod are divided into a plurality of parts by the partition plate when entering the elution area, and are eluted in a plurality of mutually independent branch pipes respectively so as to finish the process of separately eluting the magnetic beads; then, the solution eluted by the branch pipes which are independent from each other is contrasted and analyzed to remove error influence factors, so that the accuracy of the extraction result is improved.

Drawings

Fig. 1 is a schematic structural diagram of the present invention.

Fig. 2 is a schematic structural view of the first separator plug.

Fig. 3 is a schematic structural view of the second separator plug.

Detailed Description

The following describes in detail embodiments of the present invention with reference to the drawings.

Example 1

A multiheaded integrated nucleic acid extraction cuvette as shown in FIG. 1, which comprises a main tube 1; the inner cavity of the main pipe 1 is sequentially provided with a cracking area 2, a washing area 4 and an elution area 6 from top to bottom; hydrophobic sealing layers are respectively arranged between the cracking zone 2 and the washing zone 4 and between the washing zone 4 and the elution zone 6 for separation, and magnetic bead washing liquid and nucleic acid eluent are respectively arranged in the washing zone 4 and the elution zone 6; the inner cavity of the main pipe 1 is also provided with a magnetic track 7 extending from the cracking zone 2 to the elution zone 6, one end of the magnetic track 7 is closed, and the other end of the magnetic track is provided with an opening for the magnetic rod to extend into; a plurality of partition plates 61 are arranged in the inner cavity of the elution area 6, and the radial two ends of the partition plates 61 are respectively connected with the inner wall of the cavity of the elution area 6 and the outer wall of the magnetic track 7, so that the elution area 6 is divided into a plurality of mutually independent branch pipes 62; two adjacent partition boards 61 are distributed oppositely, the inner wall of the cavity of the elution area 6 and the outer wall of the magnetic track 7 are distributed oppositely to form four walls of the branch pipe 62, and the bottom surface of the elution area 6 is connected with the bottom ends of the four walls of the branch pipe 62 to form the bottom surface of the branch pipe 62. In this way, the hydrophobic sealing layer separates the cell lysis solution in the lysis zone 2 from the magnetic bead washing solution in the washing zone 4, and separates the magnetic bead washing solution in the washing zone 4 from the nucleic acid eluent in the elution zone 6; after a magnetic rod is placed into an inner hole of the magnetic track 7 from the top opening of the magnetic track 7, magnetic beads in the cracking area 2 are sucked and attached to the outer wall of the magnetic track 7 by using the magnetic force of the magnetic rod; the magnetic bar moves downwards to drive the magnetic beads to move downwards to pass through the hydrophobic sealing layer to enter the washing area 4 and the elution area 6 respectively for washing and elution so as to complete the extraction of nucleic acid; in this way, the magnetic beads are attached to the outer wall of the magnetic track 7 and move along with the magnetic rod, the outer wall of the whole magnetic track 7 can be used for moving the magnetic beads without being limited by the number of magnetic bead channels, a large number of magnetic beads can be driven at one time, and the nucleic acid extraction efficiency is improved; meanwhile, the magnetic beads are uniformly distributed on the outer wall of the magnetic track with larger area and are not stacked, so that the magnetic beads are more stable in movement. In this way, the magnetic beads on the magnetic rod are separated into a plurality of parts by the partition board 61 when entering the elution zone 6, and are eluted in a plurality of mutually independent branch pipes 62 respectively, thereby completing the separate elution process of the magnetic beads; then, the solution eluted by the branch pipes 62 which are independent of each other is contrasted and analyzed to eliminate error influence factors, so that the accuracy of the extraction result is improved.

In addition, the utility model also discloses an integrated nucleic acid extraction and detection device, which comprises the integrated nucleic acid extraction test tube and a magnetic rod, wherein the outer wall of the magnetic rod is matched with the inner hole of the magnetic track 7.

The inner cavity of the main pipe 1 is provided with a first separating plug 3 and a second separating plug 5; annular gaps are formed between the outer wall of the magnetic track 7 and the first separating plug 3 and the second separating plug 5; the hydrophobic sealing layers are respectively positioned in the first separating plug 3 and the second separating plug 5 and fill and seal the inner cavity of the main pipe 1 at the positions.

The hydrophobic sealing layer is a paraffin or liquid-phase hydrophobic layer; the first and second partition plugs 3 and 5 are both of an elastic material. The bottom ends of the branch pipes 62 are in the shape of independent cylinders. A plurality of branch pipes 62 are distributed along the center of the main pipe 1 in a circumferential shape; the axial central axis of the magnetic track 7 is axially coincident with the axial central axis of the main pipe 1.

As shown in fig. 2, the first separator plug 3 includes a first plug body 31, a first circular arc surface 32, a first prismatic surface 33, and a first center hole 34; the upper end of the inner wall of the main pipe 1 is in a circular arc shape, and the lower end of the inner wall of the main pipe 1 is in a hexagonal prism shape; the first arc surface 32 is arc-shaped and is in interference fit with the arc inner wall at the upper end of the main pipe 1, and the first prismatic surface 33 is hexagonal prism-shaped and is in interference fit with the hexagonal prism inner wall at the lower end of the main pipe 1; the first central hole 34 is penetrated by the track 7 and the gap with the outer wall of the track 7 is filled and sealed by a hydrophobic sealing layer. The first arc surface 32 and the first prismatic surface 33 are respectively in interference fit with the inner wall of the main pipe 1, so that the first separating plug 3 is clamped and positioned to the inner wall of the main pipe 1.

As shown in fig. 3, the second separator plug 5 includes a second plug body 51, a second prismatic surface 52, and a second center hole 53; the inner wall of the main pipe 1 is also provided with a second step surface 11; the bottom surface of the second plug body 51 in the horizontal direction is in contact with the second step surface 11 in the horizontal direction; the second prismatic surface 52 is in interference fit with the inner wall of the hexagonal prism at the lower end of the main pipe 1, the second center hole 53 is penetrated by the magnetic track 7, and the gap between the outer wall of the magnetic track 7 and the second center hole is filled and sealed by the hydrophobic sealing layer. The second step surface 11 is in contact with the bottom surface of the second plug body 51 to axially limit the second separating plug 5, and the second prismatic surface 52 is in interference fit with the inner wall of the main pipe 1 to radially limit the second separating plug 5; so that the second separating plug 5 can be clamped on the inner wall of the main pipe 1 by using the elasticity of the second separating plug and the limit of the inner wall of the main pipe 1.

As shown in fig. 1, the top end of the main pipe 1 is provided with a pipe cover 18, and the pipe cover 18 is screwed with the external thread of the outer wall of the main pipe 1 through the internal thread to prevent external pollutants from entering the main pipe 1; the tube cover 18 is provided with a positioning hole 181 penetrating vertically, and the positioning hole 181 is matched with the outer wall of the magnetic track 7 so as to further limit the upper part of the magnetic track 7 and prevent the upper part from deviating.

When in use, 1) firstly, a lysis solution and magnetic beads are preset in the lysis zone 2, the tube cover 18 is unscrewed, then sample cells to be detected are added into the lysis zone 2, and the sample cells are lysed by the lysis solution, release complete nucleic acid and are mutually combined with the magnetic beads; or directly adding the solution containing the sample cells to be detected and the magnetic beads after the lysis to the lysis zone 2; 2) inserting a magnetic rod into the cracking zone 2 from the opening of the magnetic track 7 and moving downwards to the washing zone 4, wherein the magnetic rod attracts magnetic beads to adhere to the outer wall of the magnetic track 7 and moves downwards to the washing zone 4 along with the magnetic rod through a hydrophobic sealing layer in the first separation plug 3; when the hydrophobic sealing layer is paraffin, the main pipe 1 is heated to melt the paraffin; 3) after removing some impurities such as protein and inorganic salt in the nucleic acid combined with the magnetic beads by the magnetic bead washing solution, the magnetic rod drives the magnetic beads to continuously move downwards to pass through the hot-melted hydrophobic sealing layer in the second separating plug 5 to the elution area 6 and respectively enter a plurality of different branch pipes 62; 4) contacting the nucleic acid eluent with the washed magnetic beads to elute the nucleic acids combined on the magnetic beads into the nucleic acid eluents of different branch pipes 62; 5) the nucleic acid is subjected to PCR or isothermal amplification reaction in the nucleic acid eluent, and a molecular beacon is introduced into the nucleic acid eluent, so that the PCR or isothermal amplification reaction can be observed in real time by external equipment through fluorescent quantitative detection, and the quantity of the sample nucleic acid is obtained by analyzing a fluorescence value, so that the detection of the sample nucleic acid is realized; then, the detection results of the nucleic acid eluents eluted by the magnetic beads in the different branch pipes 62 are respectively compared to complete the integrated nucleic acid extraction and detection process.

Claims

1. A multi-headed integrated nucleic acid extraction cuvette comprising a main tube (1); the inner cavity of the main pipe (1) is sequentially provided with a cracking area (2), a washing area (4) and an elution area (6) from top to bottom; hydrophobic sealing layers are respectively arranged between the lysis zone (2) and the washing zone (4) and between the washing zone (4) and the elution zone (6) for separating, and magnetic bead washing liquid and nucleic acid eluent are respectively arranged in the washing zone (4) and the elution zone (6); the device is characterized in that a magnetic track (7) extending from the cracking zone (2) to the elution zone (6) is further arranged in the inner cavity of the main pipe (1), one end of the magnetic track (7) is closed, and the other end of the magnetic track is provided with an opening for the magnetic rod to extend into; a plurality of partition plates (61) are arranged in the inner cavity of the elution area (6), and the partition plates (61) are respectively connected with the inner wall of the cavity of the elution area (6) and the outer wall of the magnetic track (7) at two radial ends, so that the elution area (6) is divided into a plurality of mutually independent branch pipes (62); two adjacent partition plates (61) are distributed oppositely, the inner wall of the cavity of the elution area (6) and the outer wall of the magnetic track (7) are distributed oppositely to form four walls of the branch pipe (62), and the bottom surface of the elution area (6) is connected with the bottom ends of the four walls of the branch pipe (62) to form the bottom surface of the branch pipe (62).

2. The multi-tip integrated nucleic acid extraction cuvette according to claim 1, wherein the inner cavity of the main tube (1) is provided with a first partition plug (3) and a second partition plug (5); annular gaps are formed between the outer wall of the magnetic track (7) and the first separating plug (3) and the second separating plug (5); the hydrophobic sealing layer is respectively positioned in the first separating plug (3) and the second separating plug (5) and fills and seals the inner cavity of the main pipe (1) at the position.

3. The multi-headed integrated nucleic acid extraction cuvette of claim 2, wherein the hydrophobic sealing layer is a paraffin or a liquid-phase hydrophobic layer; the first separating plug (3) and the second separating plug (5) are made of elastic materials.

4. The multiple-tip integrated nucleic acid extraction cuvette according to claim 1, wherein the bottom ends of the branch tubes (62) have mutually independent cylindrical shapes.

5. The multiple-headed integrated nucleic acid extraction cuvette according to claim 1, wherein the plurality of branch tubes (62) are circumferentially distributed along the center of the main tube (1); the axial central axis of the magnetic track (7) is axially coincident with the axial central axis of the main pipe (1).

6. The multi-tip integrated nucleic acid extraction cuvette according to claim 2, wherein the first separation plug (3) comprises a first plug body (31), a first circular arc surface (32), a first prism surface (33) and a first center hole (34); the upper end of the inner wall of the main pipe (1) is in a circular arc shape, and the lower end of the inner wall of the main pipe (1) is in a hexagonal prism shape; the first arc surface (32) is in an arc shape and is in interference fit with the arc inner wall at the upper end of the main pipe (1), and the first prismatic surface (33) is in a hexagonal prism shape and is in interference fit with the hexagonal prism inner wall at the lower end of the main pipe (1); the first central hole (34) is penetrated by the magnetic track (7) and a gap between the first central hole and the outer wall of the magnetic track (7) is filled and sealed by a hydrophobic sealing layer.

7. The multi-tip integrated nucleic acid extraction cuvette according to claim 2, wherein the second partition plug (5) comprises a second plug body (51), a second prism face (52) and a second center hole (53); the inner wall of the main pipe (1) is also provided with a second step surface (11); the bottom surface of the second plug body (51) in the horizontal direction is in contact with the second step surface (11) in the horizontal direction; the second prismatic surface (52) is in interference fit with the inner wall of the hexagonal prism at the lower end of the main pipe (1), the second central hole (53) is penetrated by the magnetic track (7), and a gap between the second central hole and the outer wall of the magnetic track (7) is filled and sealed by a hydrophobic sealing layer.

8. The multi-head integrated nucleic acid extraction test tube according to claim 1, wherein the top end of the main tube (1) is provided with a tube cover (18), and the tube cover (18) is screwed with the external thread of the outer wall of the main tube (1) through internal threads; the pipe cover (18) is provided with a positioning hole (181) which vertically penetrates through, and the positioning hole (181) is matched with the outer wall of the magnetic track (7).

9. An integrated nucleic acid extraction and detection device, comprising an integrated nucleic acid extraction cuvette according to any one of claims 1 to 8 and a magnetic rod, wherein the outer wall of the magnetic rod is fitted into the inner hole of the magnetic track (7).