CN113621512A - Full-automatic nucleic acid extraction, amplification and detection integrated reaction tube and column type detection card - Google Patents
Full-automatic nucleic acid extraction, amplification and detection integrated reaction tube and column type detection card Download PDFInfo
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- CN113621512A CN113621512A CN202110314559.1A CN202110314559A CN113621512A CN 113621512 A CN113621512 A CN 113621512A CN 202110314559 A CN202110314559 A CN 202110314559A CN 113621512 A CN113621512 A CN 113621512A
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6806—Preparing nucleic acids for analysis, e.g. for polymerase chain reaction [PCR] assay
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6844—Nucleic acid amplification reactions
Abstract
The invention relates to the field of in-vitro diagnosis nucleic acid detection, and provides a full-automatic nucleic acid extraction, amplification and detection integrated reaction tube and a column type detection card. The integrated reaction tube sequentially comprises a right-angle circular truncated cone section, a cylindrical section and a right-angle conical section along the direction from the opening part to the bottom part, wherein the right-angle circular truncated cone section forms a cracking area, the cylindrical section forms a washing area, the right-angle conical section forms an elution and amplification area, and the right-angle side of the right-angle circular truncated cone section and the right-angle side of the right-angle conical section are positioned on the same side and flush with the side part of the cylindrical section. The column type detection card is the integrated reaction tube filled with lysis solution, washing solution and reaction solution, wherein the liquids are separated by phase change separators. The reaction tube can complete the whole extraction, amplification and detection process in one tube, and has the characteristics of simple structure, convenient production, simple magnet movement structure of matched detection equipment and the like.
Description
Technical Field
The invention relates to the field of in-vitro diagnosis nucleic acid detection, in particular to a full-automatic nucleic acid extraction, amplification and detection integrated reaction tube and a column type detection card adopting the reaction tube.
Background
The process from nucleic acid extraction to amplification detection comprises the steps of cracking, washing, elution, amplification and the like, the mainstream technology at present needs to be carried out step by step in a standard laboratory, the whole process is easy to pollute the sample nucleic acid, the operation difficulty is high, the requirement on operators is high, and the popularization of the nucleic acid detection technology is not facilitated.
Chinese patent application publication No. CN111876468A discloses a full-automatic nucleic acid detection test tube, which comprises: the device comprises a main pipe body, an annular pipe groove, a conical pipe body and a turnover cover; the main pipe body, the annular pipe groove and the conical pipe body are communicated through the side face; the main pipe body is a cracking liquid area; the annular pipe groove is a cleaning area; the conical tube is a reaction zone. The detection principle is as follows: the outside magnet sets up in the intercommunication mouth department of being responsible for body, annular tube groove, and when the sample in being responsible for the internal schizolysis of body and accomplish, the magnetic bead gathers under the attraction of outside magnet, and outside magnet rotates a week along the lateral wall of annular tube groove, drives the magnetic bead and passes hydrophobic substance from being responsible for the body, and the impurity of magnetic bead is washed away to the washing liquid through annular tube groove, passes hydrophobic substance entering toper body and carries out nucleic acid reaction.
In the invention patent application, the lysis liquid tube, the cleaning liquid tube and the reaction liquid tube are connected together, the whole extraction and amplification process is carried out under a closed condition, the environmental pollution is effectively avoided, the extraction, amplification and detection are fully automatically completed through equipment in the whole experiment process, and the experiment operation difficulty is greatly simplified. However, the following drawbacks still exist: 1. the structure of the detection test tube is complex, which is not beneficial to production. Because the test tube includes the annular tube groove, leads to the structure complicacy, so be unfavorable for the production of test tube, also be unfavorable for the encapsulation of three kinds of liquid. 2. The inconsistent direction of motion of the external magnet can increase the complexity of the associated detection device. The motion trail of the magnetic beads is: the main pipe body moves downwards to enter the annular pipe groove, rotates for a circle along the annular pipe groove, and then moves downwards to enter the conical pipe body. The movement of the magnetic beads is driven by an external magnet, so that the external magnet needs to be capable of moving up and down and also capable of making circular motion around the annular tube groove, which makes the structure of the matched detection equipment complicated.
Disclosure of Invention
The invention aims to provide a full-automatic nucleic acid extraction, amplification and detection integrated reaction tube, which solves the defects of the related technology at least to a certain extent.
In order to achieve the purpose, the technical scheme adopted by the invention is as follows:
the utility model provides a full-automatic nucleic acid extraction, amplification, detection integration reaction tube, wherein, along the direction of oral area to bottom, integration reaction tube includes right angle circular truncated cone section, cylinder section and right angle cone section in proper order, right angle circular truncated cone section constitutes the schizolysis district, the cylinder section constitutes the washing district, right angle cone section constitutes elution amplification district, the right angle side of right angle circular truncated cone section and the right angle side of right angle cone section are in same one side, and flush with the lateral part of cylinder section.
In the above-mentioned full-automatic nucleic acid extraction, amplification, detection integrated reaction tube, preferably, the right-angled circular table section, the right-angled conical section and the cylindrical section are respectively provided with a heating structure at the side opposite to the right-angled side, and the heating structure can be matched with the heat conduction surface of an external heating element in the detection process. More preferably, the heating structure is a curved surface adapted to the reaction tube.
In the above-mentioned full-automatic nucleic acid extraction, amplification, detection integrated reaction tube, preferably, the inner diameter of the cylindrical section is smaller than the average inner diameter of the right-angled truncated cone section and larger than the average inner diameter of the right-angled conical section. More preferably, the inner diameter of the cylindrical section is 2 to 10mm, and the ratio of the length of the cylindrical section to the inner diameter is greater than 5.
In the above-mentioned full-automatic integrated reaction tube for nucleic acid extraction, amplification and detection, preferably, the bottom of the integrated reaction tube comprises a semi-spherical structure, and the mouth of the integrated reaction tube comprises a flip integrally formed with the integrated reaction tube.
A column type detection card comprises a lysis solution, a washing solution, a reaction solution, a phase change partition and a reaction tube; the reaction tube is the integrated reaction tube described in any one of the above; the device comprises an integrated reaction tube, a pyrolysis zone, a washing zone, an elution amplification zone, a washing zone, a reaction zone, a.
Compared with the prior art, the invention has at least the following beneficial effects:
the integrated reaction tube adopting the three-section structure has no other structures inside the tube body, and is simple in structure and convenient to produce. Meanwhile, the washing movement direction of the magnetic bead-nucleic acid complex in the experiment process is the same as the conveying direction of the magnetic bead-nucleic acid complex between adjacent sections, so that an external magnet for driving the magnetic bead to move only needs to move in one direction (along the axial direction of the reaction tube), and the magnetic bead-nucleic acid complex has the characteristic of simple movement structure of the magnet of the required matched detection equipment.
The design of right angle round platform and right angle circular cone make the schizolysis zone and elution amplification district form single face contraction structure respectively on the one hand, are favorable to improving the capacity of schizolysis zone and the quantity of saving reaction liquid, and on the other hand is in the right angle side of the two with one side and flush with the lateral part of cylinder section for in the whole transfer process of magnetic bead, outside magnet keeps unanimous to the suction of magnetic bead, can avoid the magnetic bead loss.
By adopting the integrated reaction tube, the whole extraction and amplification process is completed in one reaction tube, the operation is simple, the environmental pollution can be effectively avoided, and the full-automatic nucleic acid extraction, amplification and detection can be realized.
The heating structure extends to the cracking zone (right-angled circular table section), and can heat the cracking zone, thereby being beneficial to cracking of some complex samples.
The specification of the cylindrical section of the magnetic bead-nucleic acid complex ensures that the magnetic bead-nucleic acid complex has enough time to remove residual lysis solution and samples on the surface, can reduce the dosage of washing solution and improve the stability of the whole filling liquid layered system.
The semicircle sphere structure of bottom can be to detecting the light condensation, is favorable to improving and detects the precision.
Drawings
FIG. 1 is a schematic structural view of an embodiment of an integrated reaction tube;
FIG. 2 is a sectional view thereof;
reference numerals: 1. a cover is turned; 2. a mouth; 3. a right-angle side; 4. a semi-spherical structure; 5. a right-angled circular truncated cone section; 6. a cylindrical section; 7. a right angle conical section; 8. a heating structure; 9. a cleavage zone; 10. a washing zone; 11. eluting the amplification zone.
Detailed Description
The invention is further illustrated with reference to the following figures and examples.
The first embodiment is as follows:
the integrated reaction tube is a test tube used in vitro diagnosis nucleic acid detection, and is a test tube integrating a cracking container, a washing container, an amplification reaction container and a detection container.
Referring to fig. 1 to 2, along the direction from the mouth 2 to the bottom, the integrated reaction tube sequentially comprises a right-angle circular truncated cone section 5, a cylindrical section 6 and a right-angle circular cone section 7, wherein the right-angle circular truncated cone section 5 forms a cracking zone 9, the cylindrical section 6 forms a washing zone 10, the right-angle circular cone section 7 forms an elution and amplification zone 11, and the right-angle side 3 of the right-angle circular truncated cone section 5 and the right-angle side 3 of the right-angle circular cone section 7 are on the same side (left side in fig. 1) and flush with the side of the cylindrical section 6, and it can be seen that the left side of the tube body is a straight line.
In fig. 1, the right side portion of the right truncated cone 5, the right side portion of the right conical section 7, and the right side portion of the cylindrical section 6, i.e., the side portion opposite to the right side 3 (left side in fig. 1), are provided with a heating structure 8, and the heating structure 8 may be any surface structure capable of being fitted with the heat conduction surface of an external heating body, preferably an arc surface adapted to the reaction tube. This design makes in the testing process, and the right flank portion of right angle circular platform section 5, the right flank portion of right angle cone section 7 and the right flank portion of cylinder section 6 can be identical with the heat-conduction surface of outside heat-generating body, give the reaction zone that corresponds, accurate control reaction temperature with the high-efficient conduction of heat energy of outside heat-generating body.
Further, the inner diameter of the cylindrical section 6 is smaller than the average inner diameter of the right-angled frustum section 5 and larger than the average inner diameter of the right-angled cone section 7. The inner diameter of the cylindrical section 6 is preferably 2 to 10mm, the ratio between the length and the inner diameter of the cylindrical section 6 being greater than 5.
Further still include semicircle sphere structure 4 in the bottom of integration reaction tube, the oral area 2 of integration reaction tube still include with integration reaction tube integrated into one piece's flip 1.
Example two:
a column type detection card based on the reaction tube, which is used for in vitro diagnosis nucleic acid detection. The column type detection card comprises a lysis solution, a washing solution, a reaction solution and the integrated reaction tube; the lysis solution, the washing solution and the reaction solution are correspondingly contained in a lysis area 9, a washing area 10 and an elution amplification area 11 of the integrated reaction tube, phase-change separators are respectively arranged between the lysis solution and the washing solution and between the washing solution and the reaction solution, and the mouth part 2 of the integrated reaction tube is sealed by the flip cover 1. The phase change spacer is a material which is in a solid state at the storage temperature and is converted into a liquid state at the reaction temperature, preferably a material which is immiscible with the lysis solution, the washing solution and the reaction solution, such as paraffin.
The preparation method of the column type detection card comprises the following steps: the column type detection card is formed by containing reaction liquid in an elution amplification area 11 of the integrated reaction tube, then loading a layer of phase change partition, then loading washing liquid into a washing area 10, then loading a layer of phase change partition above the washing liquid, then loading lysis liquid into a cracking area 9, and sealing a sealing part 2 by a flip cover 1.
The method for detecting nucleic acid by using the column-type detection card is as follows:
before reaction, adding a sample and magnetic beads into a lysis zone 9, and lysing for a period of time to combine the magnetic beads with nucleic acids to form a magnetic bead-nucleic acid complex;
driving the external magnet to move from top to bottom on the left side of the reaction tube in FIG. 1, and attracting the magnetic bead-nucleic acid complex to move from top to bottom synchronously;
when moving to the amplification reaction zone (i.e., elution amplification zone 11), the magnetic bead-nucleic acid complex is separated while staying in the zone for a certain period of time;
driving the external magnet to move upwards, and attracting the magnetic beads to move upwards synchronously until the magnetic beads are approximately positioned in the middle of the reaction tube;
controlling an external heating element corresponding to the amplification reaction zone of the reaction tube to work, and heating the amplification reaction zone;
in the reaction process, the optical detection probe detects the fluorescent signal at the bottom of the reaction tube to obtain a detection result, and all parts in the detection device reset after the reaction is finished.
It can be seen from the above that this integral type reaction tube is syllogic structure, and there is not other structures inside the body, therefore has simple structure, the advantage of the production of being convenient for. In addition, in the detection process, the washing movement direction of the magnetic bead-nucleic acid complex is the same as the conveying direction of the magnetic bead-nucleic acid complex between adjacent sections, and the magnetic bead-nucleic acid complex and the conveying direction of the magnetic bead-nucleic acid complex are both in the up-down direction, so that an external magnet for driving the magnetic bead to move only needs to move in one direction, and the magnetic bead-nucleic acid complex has the characteristic of simple magnet movement structure of required matched detection equipment. By adopting the integrated reaction tube, the whole extraction and amplification process is completed in one reaction tube, so that the environmental pollution can be effectively avoided, and the full-automatic nucleic acid extraction, amplification and detection can be realized. The hemispheric structure 4 at the bottom can condense detection light, and is favorable for improving the detection precision.
In addition, the right angle round platform design and the right angle circular cone design of above-mentioned reaction tube make lysis zone 9 and elution amplification zone 11 form single face contraction structure respectively on the one hand, are favorable to improving the capacity of lysis zone 9 and the quantity of sparingly reaction liquid, and on the other hand the two right angle side 3 is located same side, and flushes with the lateral part of cylinder section 6 for in the whole transfer process of magnetic bead, the outside magnet keeps unanimous to the suction of magnetic bead, can avoid the magnetic bead loss. The heating structure 8 extends to the cracking zone 9 (right-angled circular platform section 5), and can heat the cracking zone 9, thereby facilitating the cracking of some complex samples. The cylindrical section 6 is designed into a slender structure, and the specification of the cylindrical section enables the magnetic bead-nucleic acid complex to have enough time to remove residual lysis solution and samples on the surface, can reduce the using amount of washing solution, and improves the stability of the whole filling liquid layered system.
The present invention has been described in detail with reference to the specific embodiments, and the detailed description is only for the purpose of helping those skilled in the art understand the present invention, and is not to be construed as limiting the scope of the present invention. Various modifications, equivalent changes, etc. made by those skilled in the art under the spirit of the present invention shall be included in the protection scope of the present invention.
Claims (7)
1. The full-automatic nucleic acid extraction, amplification and detection integrated reaction tube is characterized by comprising a right-angle circular truncated cone section, a cylindrical section and a right-angle conical section in sequence along the direction from an opening part to the bottom, wherein the right-angle circular truncated cone section forms a cracking area, the cylindrical section forms a washing area, the right-angle conical section forms an elution amplification area, and the right-angle side of the right-angle circular truncated cone section and the right-angle side of the right-angle conical section are positioned on the same side and flush with the side part of the cylindrical section.
2. The integrated reaction tube for automatic nucleic acid extraction, amplification and detection according to claim 1, wherein the right-angled truncated cone section, the right-angled cone section and the cylindrical section are respectively provided with a heating structure at the side opposite to the right-angled side, and the heating structure can be matched with the heat conduction surface of an external heating element during detection.
3. The integrated reaction tube for full-automatic nucleic acid extraction, amplification and detection according to claim 2, wherein the heating structure is a cambered surface adapted to the reaction tube.
4. The integrated reaction tube for automatic nucleic acid isolation, amplification and detection according to claim 1, wherein the inner diameter of the cylindrical section is smaller than the average inner diameter of the right-angled truncated cone section and larger than the average inner diameter of the right-angled cone section.
5. The integrated reaction tube for automatic nucleic acid extraction, amplification and detection according to claim 4, wherein the inner diameter of the cylindrical section is 2 to 10mm, and the ratio of the length of the cylindrical section to the inner diameter is greater than 5.
6. The integrated reaction tube for automatic nucleic acid extraction, amplification and detection according to claim 1, wherein the bottom of the integrated reaction tube comprises a semi-spherical structure, and the mouth of the integrated reaction tube comprises a flip integrally formed with the integrated reaction tube.
7. The utility model provides a column type detects card which characterized in that:
comprises lysis solution, washing solution, reaction solution, a phase change separator and a reaction tube;
the reaction tube is an integrated reaction tube according to any one of claims 1 to 6;
the device comprises an integrated reaction tube, a pyrolysis zone, a washing zone, an elution amplification zone, a washing zone, a reaction zone, a.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110314559.1A CN113621512A (en) | 2021-03-24 | 2021-03-24 | Full-automatic nucleic acid extraction, amplification and detection integrated reaction tube and column type detection card |
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| CN202110314559.1A CN113621512A (en) | 2021-03-24 | 2021-03-24 | Full-automatic nucleic acid extraction, amplification and detection integrated reaction tube and column type detection card |
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| CN113621512A true CN113621512A (en) | 2021-11-09 |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN116855367A (en) * | 2023-09-04 | 2023-10-10 | 泰州蕾灵百奥生物科技有限公司 | Detection device and detection method for detecting multiple pathogenic genes |
| CN117282482A (en) * | 2023-11-23 | 2023-12-26 | 中国科学院空天信息创新研究院 | Method for preparing test tube and method for using test tube |
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2021
- 2021-03-24 CN CN202110314559.1A patent/CN113621512A/en active Pending
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN116855367A (en) * | 2023-09-04 | 2023-10-10 | 泰州蕾灵百奥生物科技有限公司 | Detection device and detection method for detecting multiple pathogenic genes |
| CN117282482A (en) * | 2023-11-23 | 2023-12-26 | 中国科学院空天信息创新研究院 | Method for preparing test tube and method for using test tube |
| CN117282482B (en) * | 2023-11-23 | 2024-02-13 | 中国科学院空天信息创新研究院 | Method for preparing test tube and method for using test tube |
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