CN113150956A - One-stop reactor and nucleic acid detection analyzer - Google Patents
One-stop reactor and nucleic acid detection analyzer Download PDFInfo
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- CN113150956A CN113150956A CN202110444091.8A CN202110444091A CN113150956A CN 113150956 A CN113150956 A CN 113150956A CN 202110444091 A CN202110444091 A CN 202110444091A CN 113150956 A CN113150956 A CN 113150956A
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Abstract
The invention relates to a nucleic acid detection technology, in particular to a one-stop reactor and a nucleic acid detection analyzer, wherein the one-stop reactor comprises a tube cover and a reaction tube, and the tube cover is provided with a sample adding hole and a reagent adding hole; a reagent storage tube is arranged in the reagent adding hole, a second reagent is arranged in the reagent storage tube, and a sealing film for sealing the sample adding hole and the reagent adding hole is arranged on the tube cover; the reaction tube is internally provided with a reaction cavity; the tube cap and the reaction tube are connected in a matched mode to enable the reaction cavity to be in a sealed state, and sealing films are arranged at openings of two ends of the reagent adding hole. The invention solves the problems of large volume, complex internal structure, low heating rate, long detection time, large loss in the nucleic acid extraction process, easy pollution and the like of the kit.
Description
Technical Field
The invention relates to a nucleic acid detection technology, in particular to a one-stop reactor and a nucleic acid detection analyzer.
Background
The nucleic acid detection technology is a molecular biological detection method, and comprises the steps of extracting nucleic acid fragments (DNA or RNA) from a sample, then carrying out replication and amplification on the nucleic acid fragments through polymerase chain reaction, and finally detecting the nucleic acid, thereby determining the specific category or source of the nucleic acid in the sample.
The nucleic acid detection method is simple and easy to implement, has extremely wide application, and can obtain an accurate result only by a small amount of samples. During detection, a sample needs to be taken firstly, and is usually collected into a sampling tube by using a swab, then is extracted from the sampling tube to a kit, and finally is put into an instrument for reaction.
In the nucleic acid extraction and amplification process, reagents such as a first reagent and a second reagent need to be added in steps to be mixed with a sample, which also increases the amount of operation.
Chinese patent CN201520594082.7 discloses a novel fluorescent quantitative PCR kit, which integrates reagent solutions required by PCR reaction together, and each reagent is connected through a pipe, and the piston valve is used to realize the on-off of each pipeline, so as to simplify the complex polymerase chain reaction. In the patent, the reagent kit is large in size due to the use of a plurality of piston valves, the heating rate is seriously influenced, and the detection time is prolonged. And the kit can not directly sample through the swab, so that sample pollution or loss can be caused in the operation process, and the detection accuracy is reduced.
Disclosure of Invention
The invention provides a one-stop reactor and a nucleic acid detection analyzer, which aim to solve the problems of large volume, complex internal structure, low heating rate, long detection time, high loss in the nucleic acid extraction process, easy pollution and the like of a kit.
In order to achieve the purpose, the invention adopts the following technical means:
the device comprises a one-stop reactor, a reaction cavity is arranged in the one-stop reactor, and a sample adding hole and a reagent adding hole which are used for communicating the reaction cavity are formed in the one-stop reactor; a reagent storage tube is arranged in the reagent adding hole, and a second reagent is arranged in the reagent storage tube; the one-station reactor is provided with a sealing film for sealing the sample adding hole and the reagent adding hole.
The one-stop reactor comprises a tube cover and a reaction tube, wherein a reaction cavity is arranged in the reaction tube, and the tube cover and the reaction tube are connected in a matching way to enable the reaction cavity to be in a sealed state; the sample adding hole and the reagent adding hole are arranged on the tube cover.
The cross section of the tube cover is circular or square.
The outer wall of the pipe cover is provided with a first bulge. The first bulge is used for being matched with the mounting hole position of the instrument so as to position the one-stop reactor.
The lower half part of the reaction tube is flat, and the lower half part of the reaction tube is provided with a transparent detection window.
The detection window is positioned at the bottom of the reaction tube.
The first half cross section of reaction tube is circular or rectangle, the first half and the tube cap cooperation of reaction tube are connected, the reaction tube opening part is equipped with the seal membrane and makes the one end in application of sample hole and reagent hole sealed, it makes the other end in application of sample hole and reagent hole sealed to be equipped with the seal membrane on the upper surface of tube cap.
The sealing membrane is an easily-punctured membrane. (reference herein to a pierceable membrane means that the reagent storage tube is capable of piercing the sealing membrane without interfering with normal use of the reagent storage tube.)
The one-station reactor is provided with a flat reaction part, and the reaction part is provided with a transparent detection window.
And a rubber sealing plug is arranged in the sampling hole.
And a second bulge is arranged on the inner wall of the sampling hole, and a second groove matched with the second bulge is formed in the rubber sealing plug.
The reagent storage tube is movably arranged in the reagent adding hole.
The reagent storage tube is arranged in the reagent adding hole in a sliding mode.
The reagent storage tube is tubular, one end of the reagent storage tube is closed, and the other end of the reagent storage tube is open.
The open end of the reagent storage tube has a beveled edge.
The closed end of the reagent storage tube is provided with a first groove.
The reagent storage tube is also internally provided with a gravity ball, the second reagent is a freeze-dried ball, and the gravity ball is arranged above the freeze-dried ball.
The gravity ball is made of high polymer materials.
Still have the elastic component in the reagent deposits the pipe, the second reagent is the freeze-drying ball, the elastic component lower extreme offsets with the freeze-drying ball, the upper end offsets with the reagent deposits the pipe.
The elastic piece is a plastic spring.
The plastic spring is pre-pressed in the reagent storage tube, so that the vibration of the freeze-drying ball in the transportation process can be reduced, and when the sealing film on the reaction tube is punctured, the force for ejecting the freeze-drying ball can be given, so that the freeze-drying ball can fall into the solution more smoothly.
And sealing films are arranged at openings at two ends of the sample adding hole and openings at two ends of the reagent adding hole.
The sampling hole and the reagent hole are both circular through holes.
The reaction chamber has a first reagent therein.
A nucleic acid detection analyzer comprises a temperature control module, a detection module and a one-stop reactor, wherein the temperature control module controls the temperature in a reaction cavity of the one-stop reactor, and the detection module detects reaction liquid in the reaction cavity of the one-stop reactor.
The invention has the beneficial effects that:
(1) because the first reagent and the second reagent are arranged in the one-stop reactor and can respectively react with the sample, the extraction of nucleic acid and the polymerase chain reaction can be carried out in the one-stop reactor, the transfer of nucleic acid is avoided, and the cost and the complexity of the instrument are greatly reduced;
(2) the second reagent can be added under the condition that the one-station reactor is not opened, so that the operation steps are reduced, and the risk of sample pollution is reduced;
(3) the rubber sealing plug is arranged in the sample adding hole of the one-stop reactor, so that the sample is added by the injector conveniently, and the sample is automatically sealed by the elasticity of the rubber plug after being added, so that the sealing property of the reaction tube is improved; the sealing films of the sample adding hole and the reagent hole can ensure that the interior of the one-stop reactor is in a sealing state, so that the one-stop reactor is prevented from being polluted, and the detection accuracy is improved;
(4) the second bulge is matched with the second groove of the rubber sealing plug, so that the rubber sealing plug cannot slide in the sample adding hole, the puncture of the sample adding needle can be smoothly carried out, and the situation that the rubber sealing plug 55 moves to damage the sealing film is avoided;
(5) the first bulge on the outer wall of the tube cover is used for being matched with an instrument to realize positioning, is convenient to install and ensures that the one-stop reactor can be installed at a specified position for reaction and detection;
(6) because only one point of the sealing membrane contacting with the blade tip of the inclined blade is punctured, the membrane at the far end of the blade tip is not torn but is connected with the original membrane body, and the sealing membrane is ensured not to fall into the reagent;
(7) the freeze-drying ball is matched with the reagent storage tube, so that the condition that liquid reagents are easy to flow and seep is not considered, the freeze storage is invalid, the second reagents can be conveniently loaded, sealed, stored and transported, and the convenience of the one-stop reactor and the instrument is improved;
(8) the gravity ball is arranged above the freeze-drying ball, so that gaps in the reagent storage tube can be filled, and the freeze-drying ball is prevented from being cracked due to vibration in the tube in the transportation process; the gravity ball can press the freeze-drying ball under gravity, so that the freeze-drying ball can smoothly fall from the reagent storage tube, and the freeze-drying ball can be pressed after falling, so that the contact area between the freeze-drying ball and the first reagent is increased, and the reagent is fully dissolved;
(9) the lower part of the reaction tube is designed to be flat, so that the heating modules can be completely attached, and the heating module is of a thin-wall structure, so that the heating efficiency is further increased, and the detection time is finally greatly shortened;
(10) the one-stop reactor has the advantages of simple integral structure and small volume.
Drawings
FIG. 1 is a schematic structural view of the present invention;
fig. 2 is a sectional view of fig. 1.
The reference numbers in the figures denote:
5-one-stop reactor, 51-tube cover, 52-reaction tube, 53-reaction cavity, 54-reagent storage tube, 55-rubber sealing plug, 56-first protrusion, 57-gravity ball, 58-freeze-drying ball, 59-inclined blade, 510-first groove.
DETAILED DESCRIPTION OF EMBODIMENT (S) OF INVENTION
Example 1
As shown in fig. 1 and 2, the one-station reactor, which includes a tube cap 51 and a reaction tube 52,
the tube cover 51 is provided with a sample adding hole and a reagent adding hole; a reagent storage tube 54 is arranged in the reagent adding hole, a second reagent is arranged in the reagent storage tube, and a sealing film for sealing the sample adding hole and the reagent adding hole is arranged on the tube cover 51;
a reaction cavity 53 is arranged in the reaction tube 52;
the tube cap 51 and the reaction tube 52 are connected in a matching way to make the reaction chamber in a sealed state.
The cross section of the cap 51 is circular or square. The cylindrical shape is convenient to grab.
The outer wall of the tube cover is provided with a first protrusion 56. The first bulge is used for being matched with the mounting hole position of the instrument so as to position the one-stop reactor.
A rubber sealing plug 55 is mounted in the sample application hole.
The inner wall of the sample adding hole is provided with a second bulge, and the rubber sealing plug 55 is provided with a second groove matched with the second bulge.
The reagent storage tube 54 is movably installed in the reagent adding hole of the tube cap 51.
The reagent storage tube 54 is slidably disposed in the reagent addition hole of the tube cover 51.
The reagent storage tube is tubular, one end of the reagent storage tube is closed, and the other end of the reagent storage tube is open.
The open end of the reagent holding tube 54 has a beveled edge 59 and the closed end is provided with a first recess 510.
The reagent holding tube 54 also has a gravity ball 57 therein, the second reagent is a lyophilized ball 58, and the gravity ball 57 is disposed above the lyophilized ball 58.
The gravity ball is a solid with a smooth surface and is made of a high polymer material.
The gravity ball in this embodiment can be replaced by an elastic member, the second reagent is a freeze-drying ball 58, the lower end of the elastic member abuts against the freeze-drying ball, and the upper end of the elastic member abuts against the reagent storage tube.
The elastic piece is a plastic spring.
The plastic spring is pre-pressed in the reagent storage tube, so that the vibration of the freeze-drying ball in the transportation process can be reduced, and when the sealing film on the reaction tube is punctured, the force for ejecting the freeze-drying ball can be given, so that the freeze-drying ball can fall into the solution more smoothly.
The two ends of the sample adding hole are opened, and the two ends of the reagent adding hole are opened respectively with sealing films.
The sampling hole and the reagent hole are both circular through holes.
The reaction tube has a first reagent therein.
The lower half of the reaction tube 52 is flat and has a transparent detection window.
The detection window is located at the bottom of the reaction tube 52.
The first half cross section of reaction tube 52 is circular or rectangle, and the first half of reaction tube 52 is connected with the cooperation of tube cap 51, and the opening part of reaction tube 52 is equipped with the seal membrane and makes the one end in addition hole and reagent hole sealed, is equipped with the seal membrane on the upper surface of tube cap 51 and makes the other end in addition hole and reagent hole sealed.
The round or rectangular tube is convenient to process and take and place, and in addition, the round tube wall can ensure uniform thickness, and the temperature inside the reaction tube is ensured to be stabilized at a target value by heating. The upper part of the structure is wider, the reagent can be ensured to be fully contacted and reacted with the sample, the freeze-dried ball is ensured to be dissolved, and the heat conduction speed and the PCR amplification speed are ensured due to the narrower lower structure.
The sealing film is an easily-punctured film. (reference herein to a pierceable membrane means that the reagent storage tube is capable of piercing the sealing membrane without interfering with normal use of the reagent storage tube.)
The working principle of the present embodiment is explained in detail below.
During sample adding, the sealing film on the tube cover 51, the rubber sealing plug and the sealing film on the reaction tube 52 are pierced through the injector, the sample is added in the reaction cavity, and when the injector is taken out, the rubber sealing plug is automatically sealed through elasticity.
Because the reaction tube is provided with the first reagent, the sample can firstly react with the first reagent, and because the tube cover is provided with the second reagent, the second reagent can be timely added into the reaction tube to carry out the next reaction on the sample. And after the reaction is finished, the detection device detects through a detection window at the bottom of the reaction tube.
The working principle of adding the second reagent into the reaction tube is as follows: the reagent storage tube is pushed, and the sealing film on the reaction tube can be punctured by the inclined blade of the reagent storage tube, so that the second reagent enters the reagent reaction tube and is mixed with the first reagent and the sample.
The device can be provided with a mandril, the one-stop reactor is arranged in the reaction cavity, and the mandril can move to push the reagent storage tube.
The second reagent in this embodiment is stored in the form of a lyophilized pellet, which has the benefits of:
when the sealing membrane is punctured to the inclined edge of reagent storage tube, freeze-drying ball and gravity ball all can fall into in the reaction tube, and the gravity of gravity ball can guarantee that freeze-drying ball can fall and invade in the first reagent completely.
After the freeze-drying ball is completely dissolved, the instrument heats the thin wall of the lower part of the reaction tube, PCR reaction is started, and finally detection is carried out through a detection window at the bottom of the reaction tube.
The beneficial effect of this embodiment lies in:
because the first reagent and the second reagent are arranged in the one-stop reactor and can respectively react with the sample, the extraction of nucleic acid and the PCR amplification reaction can be carried out in the one-stop reactor, the transfer of nucleic acid is avoided, and the cost and the complexity of the instrument are greatly reduced;
the second reagent can be added under the condition that the one-station reactor is not opened, so that the operation steps are reduced, and the risk of sample pollution is reduced;
the rubber sealing plug is arranged in the sample adding hole of the one-stop reactor, so that the sample is added by the injector conveniently, and the sample is automatically sealed by the elasticity of the rubber plug after being added, so that the sealing property of the reaction tube is improved;
the sealing films of the sample adding hole and the reagent hole can ensure that the interior of the one-stop reactor is in a sealing state, so that the one-stop reactor is prevented from being polluted, and the detection accuracy is improved;
the second protrusion is matched with the second groove of the rubber sealing plug 55, so that the rubber sealing plug 55 is ensured not to slide in the sample adding hole, the puncture of the sample adding needle is ensured to be smoothly carried out, and the condition that the rubber sealing plug 55 is moved to damage the sealing film is also avoided.
The first bulge on the outer wall of the tube cover is used for being matched with an instrument to realize positioning and facilitate installation, and ensures that the one-stop reactor can be installed at a specified position for reaction and detection.
The reagent storage tube is used for storing and releasing a second reagent, when the second reagent is released, the blade tip of the inclined blade pierces one point of the sealing membrane, the reagent storage tube continues to move, and the edge of the inclined blade tears the sealing membrane, so that the reagent storage tube can penetrate through the sealing membrane, and the second reagent enters the reaction tube. Only one point of the sealing membrane contacted with the cutting edge of the inclined blade is punctured, and the membrane at the far end of the cutting edge is not torn and is connected with the original membrane body, so that the sealing membrane is ensured not to fall into the reagent.
The freeze-drying ball is matched with the reagent storage tube, so that the condition that liquid reagents are easy to flow and seep is not considered, the freeze preservation is invalid, the second reagents can be conveniently loaded, sealed, preserved and transported, and the convenience of the one-stop reactor and the instrument is improved.
The gravity ball is arranged above the freeze-drying ball, so that gaps in the reagent storage tube can be filled, and the freeze-drying ball is prevented from being cracked due to vibration in the tube in the transportation process. The gravity ball can press the freeze-drying ball through gravity down, makes the freeze-drying ball fall smoothly from the reagent storage tube, and can push down the freeze-drying ball after the whereabouts, increases the area of contact of freeze-drying ball and first reagent for reagent fully dissolves.
The lower part of the reaction tube is designed to be flat, so that the heating module can be completely attached, and the lower part of the reaction tube is of a thin-wall structure, so that the heating efficiency is further increased, and finally, the detection time is greatly shortened.
The one-stop reactor of the embodiment has simple integral structure and small volume.
Example 2
The one-station reactor is internally provided with a reaction cavity and is provided with a sample adding hole and a reagent adding hole; a reagent storage tube 54 is arranged in the reagent adding hole, and a second reagent is arranged in the reagent storage tube; the one-station reactor has a sealing film for sealing the sample addition hole and the reagent addition hole.
The outer wall of the one-station reactor is provided with a first protrusion 56. The first bulge is used for being matched with the mounting hole position of the instrument so as to position the one-stop reactor.
A rubber sealing plug 55 is mounted in the sample application hole.
The inner wall of the sample adding hole is provided with a second bulge, and the rubber sealing plug 55 is provided with a second groove matched with the second bulge.
The reagent holding tube 54 is movably mounted in the reagent addition hole.
The reagent storage tube 54 is slidably disposed in the reagent addition hole of the tube cover 51.
The reagent storage tube is tubular, one end of the reagent storage tube is closed, and the other end of the reagent storage tube is open.
The open end of the reagent holding tube 54 has a beveled edge 59 and the closed end is provided with a first recess 510. The first groove 510 facilitates the movement of the reagent storage tube.
The reagent holding tube 54 also has a gravity ball 57 therein, the second reagent is a lyophilized ball 58, and the gravity ball 57 is disposed above the lyophilized ball 58.
The gravity ball is a solid with a smooth surface and is made of a high polymer material.
The gravity ball in this embodiment can be replaced by an elastic member, the second reagent is a freeze-drying ball 58, the lower end of the elastic member abuts against the freeze-drying ball, and the upper end of the elastic member abuts against the reagent storage tube.
The elastic piece is a plastic spring.
The plastic spring is pre-pressed in the reagent storage tube, so that the vibration of the freeze-drying ball in the transportation process can be reduced, and when the sealing film on the reaction tube is punctured, the force for ejecting the freeze-drying ball can be given, so that the freeze-drying ball can fall into the solution more smoothly.
The two ends of the sample adding hole are opened, the two ends of the reagent adding hole are opened and are provided with sealing films,
the sampling hole and the reagent hole are both circular through holes.
The reaction chamber has a first reagent therein.
The one-stop reactor is provided with a flat reaction part, and the reaction part is provided with a transparent detection window.
This example differs from example 1 only in that: the one-stage reactor of this example was integrally molded, whereas the one-stage reactor of example 1 had a tube cap and a reaction tube. The working principle and the technical effect of the embodiment are the same as those of the embodiment 1.
Example 3
A nucleic acid detection analyzer comprises a temperature control module, a detection module and a one-stop reactor, wherein the temperature control module controls the temperature in a reaction cavity of the one-stop reactor, and the detection module detects reaction liquid in the reaction cavity of the one-stop reactor.
Claims (12)
1. A one-station reactor, comprising: a reaction cavity is formed in the one-stop reactor, and a sample adding hole and a reagent adding hole which are used for communicating the reaction cavity are formed in the one-stop reactor; a reagent storage tube is arranged in the reagent adding hole, and a second reagent is arranged in the reagent storage tube; the one-stop reactor is provided with a sealing film for sealing the sample adding hole and the reagent adding hole, and the openings at two ends of the reagent adding hole are provided with sealing films.
2. The one-station reactor of claim 1, wherein: the reaction cavity is arranged in the reaction tube, and the tube cover and the reaction tube are connected in a matching way to ensure that the reaction cavity is in a sealed state; the sample adding hole and the reagent adding hole are arranged on the tube cover.
3. The one-station reactor of claim 2, wherein: the cross section of the tube cover is circular or square;
or/and
the lower half part of the reaction tube is flat, and the lower half part of the reaction tube is provided with a transparent detection window.
4. The one-station reactor of claim 3, wherein: the detection window is positioned at the bottom of the reaction tube.
5. The one-station reactor according to claim 2, 3 or 4, wherein: the cross section of the upper half part of the reaction tube is circular or rectangular, the upper half part of the reaction tube is matched and connected with a tube cap, a sealing film is arranged at the opening of the reaction tube to seal one ends of the sample adding hole and the reagent adding hole, and the sealing film is arranged on the upper surface of the tube cap to seal the other ends of the sample adding hole and the reagent adding hole; the sealing film is an easily-punctured film; the outer wall of the pipe cover is provided with a first bulge.
6. The one-station reactor according to claim 1, 2, 3 or 4, wherein: and a rubber sealing plug is arranged in the sampling hole. And a second bulge is arranged on the inner wall of the sampling hole, and a second groove matched with the second bulge is formed in the rubber sealing plug.
7. The one-station reactor according to claim 1, 2, 3 or 4, wherein: the reagent storage tube is movably arranged in the reagent adding hole. Further, the reagent storage tube is slidably disposed in the reagent adding hole.
8. The one-station reactor according to claim 1, 2, 3 or 4, wherein: the reagent storage tube is tubular, one end of the reagent storage tube is closed, and the other end of the reagent storage tube is open. Preferably, the open end of the reagent holding tube has a beveled edge. Preferably, the closed end of the reagent storage tube is provided with a first groove.
9. The one-station reactor according to claim 1, 2, 3, 4, 5, 6, 7 or 8, wherein: the reagent storage tube is also internally provided with a gravity ball, the second reagent is a freeze-dried ball, and the gravity ball is arranged above the freeze-dried ball. Preferably, the gravity ball is made of a high polymer material.
10. The one-station reactor according to claim 1, 2, 3, 4, 5, 6, 7 or 8, wherein: still have the elastic component in the reagent deposits the pipe, the second reagent is the freeze-drying ball, the elastic component lower extreme offsets with the freeze-drying ball, the upper end offsets with the reagent deposits the pipe. Preferably, the elastic member is a plastic spring. Preferably, the sample adding hole and the reagent hole are both circular through holes.
11. The one-station reactor according to claim 1, 2, 3, 4, 5, 6, 7 or 8, wherein: the reaction chamber has a first reagent therein.
12. A nucleic acid detecting analyzer, characterized in that: the one-station reactor comprises a temperature control module, a detection module and the one-station reactor as claimed in any one of claims 1 to 11, wherein the temperature control module controls the temperature in the reaction cavity of the one-station reactor, and the detection module detects the reaction liquid in the reaction cavity of the one-station reactor.
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Cited By (2)
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| WO2023138148A1 (en) * | 2022-01-19 | 2023-07-27 | 江苏为真生物医药技术股份有限公司 | Sealing device for rapid detection, use method thereof, and application thereof |
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| CN108531377A (en) * | 2018-06-26 | 2018-09-14 | 广东省第二人民医院（广东省卫生应急医院） | It is a kind of can pre-filled reagent diaphragm type nucleic acid amplification airtight reactor tube |
| CN210367679U (en) * | 2019-07-22 | 2020-04-21 | 安图实验仪器(郑州)有限公司 | Spiral reaction tube |
| CN210367678U (en) * | 2019-07-22 | 2020-04-21 | 安图实验仪器(郑州)有限公司 | Push type reaction tube |
| CN111334425A (en) * | 2020-04-09 | 2020-06-26 | 杭州奥盛仪器有限公司 | Reaction tube for coupling double systems for nucleic acid amplification and CRISPR detection and double system coupling method |
| CN111621412A (en) * | 2020-06-17 | 2020-09-04 | 杭州奥盛仪器有限公司 | Reaction tube for two-step nucleic acid amplification and detection |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| WO2023138148A1 (en) * | 2022-01-19 | 2023-07-27 | 江苏为真生物医药技术股份有限公司 | Sealing device for rapid detection, use method thereof, and application thereof |
| CN115747037A (en) * | 2023-01-09 | 2023-03-07 | 深圳市科瑞达生物技术有限公司 | Nucleic acid analysis card and method of use thereof |
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