CN112226358B - Nucleic acid analysis card box and nucleic acid analysis equipment - Google Patents

Nucleic acid analysis card box and nucleic acid analysis equipment Download PDF

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Publication number
CN112226358B
CN112226358B CN202011435583.2A CN202011435583A CN112226358B CN 112226358 B CN112226358 B CN 112226358B CN 202011435583 A CN202011435583 A CN 202011435583A CN 112226358 B CN112226358 B CN 112226358B
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liquid storage
nucleic acid
acid analysis
storage cavity
detection
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CN112226358A (en
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李楠
张渊越
徐友春
程京
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Boao Biological Group Co ltd
Tsinghua University
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Boao Biological Group Co ltd
Tsinghua University
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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
    • 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 present invention discloses a nucleic acid analysis cartridge and a nucleic acid analysis apparatus, the nucleic acid analysis cartridge comprising: the cartridge comprises a cartridge body and a silicone membrane transfer component which is arranged on the cartridge body and can move along a set direction; the card box body is provided with at least two liquid storage cavities, auxiliary liquid storage cavities, a detection structure, a sample adding channel and an exhaust channel which are communicated with the liquid storage cavities, the auxiliary liquid storage cavities are distributed in sequence along a set direction, the auxiliary liquid storage cavities correspond to the liquid storage cavities one by one, and the auxiliary liquid storage cavity positioned at the last along the set direction can be communicated with the detection structure; the surface of the liquid storage cavity and the surface of the auxiliary liquid storage cavity are hermetically connected with the silicone film transfer part, and the silicone film transfer part moves along a set direction to enable the silicone film to sequentially communicate each liquid storage cavity and the auxiliary liquid storage cavity corresponding to the liquid storage cavity. The nucleic acid analysis card box can perform full-integrated automatic analysis of nucleic acid by adopting a silica gel membrane method, and is favorable for promoting the popularization of molecular diagnosis technology.

Description

Nucleic acid analysis card box and nucleic acid analysis equipment
Technical Field
The present invention relates to the field of in vitro diagnostic techniques, and more particularly to a nucleic acid analysis cartridge and a nucleic acid analysis apparatus.
Background
In the field of in vitro diagnosis, molecular diagnosis is a common technique, and particularly, molecular diagnosis is a diagnostic technique for clinical detection by using nucleic acid or protein as a biomarker, and provides information and decision basis for prediction, diagnosis, prevention, treatment, prognosis and outcome of diseases.
The molecular diagnosis technology relates to nucleic acid extraction, and a silica gel membrane method is one of the common technologies for nucleic acid extraction. When the silica gel membrane method is adopted for extracting nucleic acid, various reagents such as lysate, cleaning fluid, eluent and the like are needed, multi-step uncapping and liquid adding operation is needed, high-speed centrifugation is assisted, and the operation steps are complex. Clinical diagnostic laboratories to reduce the risk of contamination of multi-step nucleic acid extraction and amplification assays, multiple isolated rooms are provided for sample delivery through a delivery window. The design has higher requirement on the laboratory environment, the operation of personnel is time-consuming, and the requirements of laggard areas and on-site instant detection of medical infrastructure are difficult to meet. The fully integrated automatic molecular diagnosis system can realize the detection of 'sample input and result output' without manual intervention, and is a development trend in the field of molecular diagnosis.
In conclusion, how to adopt a silica gel membrane method to perform a fully integrated automated analysis of nucleic acids to promote the popularization of molecular diagnosis technology is a problem to be solved urgently by those skilled in the art.
Disclosure of Invention
The invention aims to provide a nucleic acid analysis card box which can fully integrate nucleic acid extraction and amplification detection based on a silica gel membrane method and promote the popularization of molecular diagnosis technology. Another object of the present invention is to provide a nucleic acid analysis apparatus having the above-mentioned nucleic acid analysis cartridge.
In order to achieve the above purpose, the invention provides the following technical scheme:
a nucleic acid analysis cartridge comprising: the cartridge comprises a cartridge body and a silicone membrane transfer component which is arranged on the cartridge body and can move along a set direction;
the card box body is provided with at least two liquid storage cavities, at least two auxiliary liquid storage cavities, a detection structure, a sample adding channel and an exhaust channel which are communicated with the liquid storage cavities, the sample adding channels and the exhaust channels are sequentially distributed along the set direction, the auxiliary liquid storage cavities correspond to the liquid storage cavities one by one, and the auxiliary liquid storage cavity positioned at the last along the set direction can be communicated with the detection structure;
the liquid storage cavity, the sample adding channel and the exhaust channel are all positioned on one side of the silicone membrane transfer component, and the auxiliary liquid storage cavity and the detection structure are all positioned on the other side of the silicone membrane transfer component;
the liquid storage cavity surface and the auxiliary liquid storage cavity surface are hermetically connected with the silicon film transfer part, the silicon film transfer part is fixed with a silicon film, and the silicon film transfer part moves along the set direction to enable the silicon film to sequentially communicate each liquid storage cavity and the auxiliary liquid storage cavity corresponding to the liquid storage cavity.
Preferably, when the silicone membrane is communicated with the liquid storage cavity and the auxiliary liquid storage cavity corresponding to the liquid storage cavity, liquid in the liquid storage cavity can enter the auxiliary liquid storage cavity corresponding to the liquid storage cavity under the centrifugal action;
the bottom end of the auxiliary liquid storage cavity is lower than the bottom end of the liquid storage cavity corresponding to the auxiliary liquid storage cavity so as to limit liquid in the auxiliary liquid storage cavity to flow back to the liquid storage cavity corresponding to the auxiliary liquid storage cavity.
Preferably, the detection structure is located at the bottom side of the auxiliary reservoir.
Preferably, the silicone membrane transfer part is in a strip shape, the silicone membrane transfer part moves linearly along the set direction, and the set direction is the length direction of the silicone membrane transfer part; the silicone film transfer component is provided with a limiting structure, and the limiting structure is used for limiting the silicone film transfer component to be separated from the card box body;
or, the silicone membrane transfer part is circular, the silicone membrane transfer part rotates around the axis of the silicone membrane transfer part, and the set direction is the circumferential direction of the silicone membrane transfer part.
Preferably, the silicone membrane transfer part is provided with a drive connection structure which can be connected with a transfer drive device, and the transfer drive device is used for driving the silicone membrane transfer part to move along the set direction;
the silicone membrane transfer component is provided with a fixing groove and a communicating hole communicated with the fixing groove, and the silicone membrane is fixed in the fixing groove through a fixing ring.
Preferably, the detection structure comprises: the sampling channel, the isolation valve, the detection cavity, the waste liquid cavity and the ventilation structure;
the detection cavity, the waste liquid cavity and the ventilation structure are all communicated with the sample feeding channel, the waste liquid cavity is positioned at the downstream of the detection cavity, the ventilation structure is positioned at the downstream of the waste liquid cavity, and the sample feeding channel is communicated with the last auxiliary liquid storage cavity positioned along the set direction; the isolation valve is arranged on the sample feeding channel, and the isolation valve can cut off and conduct the sample feeding channel.
Preferably, the vent structure comprises: the sample feeding device comprises a sample feeding channel, a ventilation channel communicated with the sample feeding channel, a ventilation fixing hole communicated with the ventilation channel, and a hydrophobic breathable film sealed in the ventilation fixing hole; wherein, the ventilation fixing hole is arranged on the card box body.
Preferably, the cartridge body comprises: the liquid storage box, the detection box, the first cover plate, the second cover plate and the third cover plate;
the liquid storage cavity, the sample adding channel and the exhaust channel are all arranged on the liquid storage box, and the first cover plate is connected with the liquid storage box in a sealing mode so as to seal the liquid storage cavity, the sample adding channel and the exhaust channel;
the auxiliary liquid storage cavity and the detection structure are arranged on the detection box, the second cover plate is in sealing connection with the detection box to seal the detection structure, and the third cover plate is in sealing connection with the detection box to seal the auxiliary liquid storage cavity;
the silicone membrane transfer part is arranged between the liquid storage box and the detection box, and the liquid storage box and the detection box are both in sealing connection with the silicone membrane transfer part;
the detection box is provided with an accommodating groove for accommodating the silicone membrane transfer part.
Preferably, the liquid storage cavity is provided with a first mounting groove, the auxiliary liquid storage cavity is provided with a second mounting groove, sealing rings are arranged in the first mounting groove and the second mounting groove, and the liquid storage box and the detection box are both connected with the silicone membrane transfer part in a sealing manner through the sealing rings;
the liquid storage box with the detection box passes through fastener fixed connection, first apron through bonding with liquid storage box sealing connection, the second apron with the third apron all through bonding with detection box sealing connection.
Preferably, the detection box, the second cover plate and the third cover plate are all transparent pieces.
According to the nucleic acid analysis card box provided by the invention, the silica gel film transfer component which can move along the set direction is arranged on the card box body, the silica gel film on the silica gel film transfer component is sequentially communicated with each liquid storage cavity and the auxiliary liquid storage cavity corresponding to the liquid storage cavity by utilizing the movement of the silica gel film transfer component, and the silica gel film can be sequentially moved to each liquid storage cavity and the auxiliary liquid storage cavity corresponding to the liquid storage cavity along with the movement of the silica gel film transfer component. Therefore, the silica gel membrane transfer component realizes transfer of the silica gel membrane, liquid in the liquid storage cavities and liquid in the auxiliary liquid storage cavities among different liquid storage cavities, so that extraction of nucleic acid is completed, amplification and detection of nucleic acid are completed in the detection structure, namely, the silica gel membrane method can be adopted for full-integration automatic analysis of nucleic acid, and popularization of molecular diagnosis technology is promoted.
Meanwhile, the nucleic acid analysis card box provided by the invention can finish nucleic acid extraction and nucleic acid amplification detection only by moving the silica gel membrane transfer part and controlling the centrifugation of the nucleic acid analysis card box, is simple to operate and further promotes the popularization of molecular diagnosis technology; the silicone film and the sample are transferred in the card box body, so that the pollution probability of the sample is reduced, and the reliability of a detection result is improved; moreover, the card box body is internally provided with a liquid storage cavity, a detection structure, a sample adding channel and an exhaust channel which are communicated with the liquid storage cavity, and then required reagents are pre-stored in the liquid storage cavity and the detection structure, so that the closed automatic extraction and amplification detection of nucleic acid is realized, the risk of interface cross contamination is avoided, and the reliability of the detection result is also improved.
Meanwhile, the liquid storage cavity and the auxiliary liquid storage cavity are distributed on two sides of the silica gel film transfer component and are sequentially distributed along the moving direction of the silica gel film transfer component, so that the space in the set direction is fully utilized, the compactness of the whole nucleic acid analysis card box is improved, the occupied area of the whole nucleic acid analysis card box is effectively reduced, and the popularization of a molecular diagnosis technology is further promoted.
Based on the nucleic acid analysis cartridge provided above, the present invention also provides a nucleic acid analysis apparatus comprising the nucleic acid analysis cartridge according to any one of the above;
the nucleic acid analysis apparatus further includes: a carrier, a centrifuge, a transfer drive, and a detection device;
the carrying component carries the nucleic acid analysis card box, the centrifugal device drives the carrying component to rotate so as to enable the nucleic acid analysis card box to do centrifugal motion, and the transfer driving device is used for driving the silica gel film transfer component to move along the set direction; the detection device is used for detecting the reaction result of the detection structure.
Preferably, the nucleic acid analysis cartridges are at least two and are sequentially arranged along the rotation direction of the carrier;
if the silica gel film transfer part is in a strip shape, the silica gel film transfer parts of at least two nucleic acid analysis card boxes are driven by the same transfer driving device; if the silica gel membrane transfer part is circular, the transfer driving device corresponds to the nucleic acid analysis card boxes one by one.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly described below, it is obvious that the drawings in the following description are only embodiments of the present invention, and for those skilled in the art, other drawings can be obtained according to the provided drawings without creative efforts.
FIG. 1 is a schematic view of a nucleic acid analysis cartridge according to an embodiment of the present invention;
FIG. 2 is an exploded view of a nucleic acid analysis cartridge according to one embodiment of the present invention;
FIG. 3 is a schematic view of a liquid storage case of a nucleic acid analysis cartridge according to an embodiment of the present invention;
FIG. 4 is a schematic view showing another aspect of a reservoir cartridge of a nucleic acid analysis cartridge according to an embodiment of the present invention;
FIG. 5 is a schematic structural view of a silicone film transfer unit in the nucleic acid analysis cartridge according to one embodiment of the present invention;
FIG. 6 is a schematic view showing a structure of a cartridge in a nucleic acid analysis cartridge according to one embodiment of the present invention;
FIG. 7 is a schematic view showing another orientation of the cartridge in the nucleic acid analysis cartridge according to the first embodiment of the present invention;
FIG. 8 is a schematic view showing another orientation of the cartridge in the nucleic acid analysis cartridge according to the first embodiment of the present invention;
FIG. 9 is a side view of a nucleic acid analysis cartridge provided in one embodiment of the present invention;
FIG. 10 is a sectional view taken along line A-A of FIG. 9;
FIG. 11 is a side view of a nucleic acid analysis cartridge provided in accordance with one embodiment of the present invention in a first state;
FIG. 12 is a front view of a nucleic acid analysis cartridge according to one embodiment of the present invention in a first state;
FIG. 13 is a side view of a nucleic acid analysis cartridge provided in accordance with one embodiment of the present invention in a second state;
FIG. 14 is a front view of a nucleic acid analysis cartridge according to a first embodiment of the present invention in a second state;
FIG. 15 is a side view of a nucleic acid analysis cartridge provided in accordance with one embodiment of the present invention in a third state;
FIG. 16 is a front view of a nucleic acid analysis cartridge according to a first embodiment of the present invention in a third state;
FIG. 17 is a side view of a nucleic acid analysis cartridge provided in accordance with one embodiment of the present invention in a fourth state;
FIG. 18 is a front view of a nucleic acid analysis cartridge in a fourth state according to one embodiment of the present invention;
FIG. 19 is a side view of a nucleic acid analysis cartridge provided in accordance with one embodiment of the present invention in a fifth state;
FIG. 20 is a front view of a nucleic acid analysis cartridge in a fifth state according to one embodiment of the present invention;
FIG. 21 is a plan view of a nucleic acid analysis cartridge provided in accordance with one embodiment of the present invention in a sixth state;
FIG. 22 is a schematic structural view of a nucleic acid analysis apparatus according to a first embodiment of the present invention;
FIG. 23 is a schematic view showing the structure of a nucleic acid analysis cartridge according to a second embodiment of the present invention;
FIG. 24 is an exploded view of a nucleic acid analysis cartridge provided in accordance with a second embodiment of the present invention;
FIG. 25 is a schematic view showing a structure of a liquid storage case in a nucleic acid analysis cartridge according to a second embodiment of the present invention;
FIG. 26 is a schematic view showing a structure of a liquid storage cassette in a nucleic acid analysis cartridge according to a second embodiment of the present invention in another orientation;
FIG. 27 is a schematic view showing the structure of a silicone film transfer member in the nucleic acid analysis cartridge according to the second embodiment of the present invention;
FIG. 28 is a schematic view showing a structure of a cartridge in a nucleic acid analysis cartridge according to a second embodiment of the present invention;
FIG. 29 is a schematic view showing a structure of a cartridge in another orientation in a nucleic acid analysis cartridge according to the second embodiment of the present invention;
FIG. 30 is a schematic view showing another direction structure of a cartridge in a nucleic acid analysis cartridge according to the second embodiment of the present invention;
FIG. 31 is a side view of a nucleic acid analysis cartridge provided in the second embodiment of the present invention;
FIG. 32 is a sectional view taken along line B-B of FIG. 31;
FIG. 33 is a side view of a nucleic acid analysis cartridge provided in accordance with a second embodiment of the present invention in a first state;
FIG. 34 is a top view of a nucleic acid analysis cartridge provided in accordance with a second embodiment of the present invention in a first state;
FIG. 35 is a side view of a nucleic acid analysis cartridge provided in the second embodiment of the present invention in a second state;
FIG. 36 is a top view of a nucleic acid analysis cartridge provided in accordance with the second embodiment of the present invention in a second state;
FIG. 37 is a side view of a nucleic acid analysis cartridge provided in the second embodiment of the present invention in a third state;
FIG. 38 is a top view of a nucleic acid analysis cartridge provided in accordance with the second embodiment of the present invention in a third state;
FIG. 39 is a side view of a nucleic acid analysis cartridge provided in accordance with the second embodiment of the present invention in a fourth state;
FIG. 40 is a top view of a nucleic acid analysis cartridge provided in accordance with the second embodiment of the present invention in a fourth state;
FIG. 41 is a side view of a nucleic acid analysis cartridge provided in the second embodiment of the present invention in a fifth state;
FIG. 42 is a top view of a nucleic acid analysis cartridge provided in accordance with a second embodiment of the present invention in a fifth state;
FIG. 43 is a plan view of a nucleic acid analysis cartridge provided in accordance with the second embodiment of the present invention in a sixth state;
FIG. 44 is a schematic structural view of a nucleic acid analysis apparatus according to a second embodiment of the present invention.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the 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 invention.
As shown in FIGS. 1 to 44, the embodiment of the present invention provides a nucleic acid analysis cartridge comprising: a cartridge body, and a silicone film transfer member 200 provided on the cartridge body and movable in a set direction.
Above-mentioned card box body is provided with stock solution chamber, supplementary stock solution chamber, detects the structure and all with the application of sample passageway 104 and the exhaust passage 105 of stock solution chamber intercommunication, and above-mentioned stock solution chamber is two at least and distributes in proper order along setting for the direction, supplementary stock solution chamber and stock solution chamber one-to-one, and it can communicate with detecting the structure to be located last stock solution chamber along setting for the direction.
The liquid storage cavity, the sample adding channel 104 and the air exhaust channel 105 are all located on one side of the silicone membrane transfer component 200, and the auxiliary liquid storage cavity and the detection structure are all located on the other side of the silicone membrane transfer component 200.
The surface of the liquid storage cavity and the surface of the auxiliary liquid storage cavity are hermetically connected with the silicone membrane transfer part 200 so as to seal reagents and waste liquid required by the reaction. It will be appreciated that the reservoir, sample addition channel 104 and vent channel 105 are all disposed on the same face. The silicone membrane transfer part 200 is fixed with a silicone membrane 700, and the silicone membrane transfer part 200 moves along a set direction to enable the silicone membrane 700 to be sequentially communicated with each liquid storage cavity and the auxiliary liquid storage cavity corresponding to the liquid storage cavity.
It can be understood that when the silicone membrane 700 communicates the liquid storage cavity and the auxiliary liquid storage cavity corresponding thereto, the liquid in the liquid storage cavity can enter the auxiliary liquid storage cavity.
The sample adding channel 104 is used for adding required reagents, and the reagents enter the liquid storage cavity through the sample adding channel 104; the air exhaust channel 105 is used for exhausting air from the liquid storage cavity when the required reagent flows into the liquid storage cavity; the sample loading channel 104 and the air exhaust channel 105 have no spatial arrangement interference so as to ensure sample loading and air exhaust.
According to the nucleic acid analysis card box provided by the embodiment of the invention, the silica gel film transfer part 200 which can move along the set direction is arranged on the card box body, and the silica gel film 700 on the silica gel film transfer part 200 is sequentially communicated with each liquid storage cavity and the auxiliary liquid storage cavity corresponding to the liquid storage cavity by utilizing the movement of the silica gel film transfer part 200, so that the silica gel film 700 can be sequentially moved to each liquid storage cavity and the auxiliary liquid storage cavity corresponding to the liquid storage cavity along with the movement of the silica gel film transfer part 200. Therefore, the silicone membrane transfer component 200 realizes transfer of the silicone membrane 700, liquid in the liquid storage cavities and liquid in the auxiliary liquid storage cavities among different liquid storage cavities, so that extraction of nucleic acid is completed, amplification and detection of nucleic acid are completed in a detection structure, namely, full-integration automatic analysis of nucleic acid can be performed by adopting a silicone membrane method, and popularization of molecular diagnosis technology is promoted.
Meanwhile, the nucleic acid analysis card box provided by the embodiment of the invention can finish nucleic acid extraction and nucleic acid amplification detection only by moving the silica gel membrane transfer part 200 and controlling the nucleic acid analysis card box to be centrifuged, is simple to operate and further promotes the popularization of molecular diagnosis technology; the silicone film 700 and the sample are transferred in the card box body, so that the pollution probability of the sample is reduced, and the reliability of the detection result is improved; moreover, the card box body is internally provided with a liquid storage cavity, a detection structure, a sample adding channel 104 and an exhaust channel 105 which are communicated with the liquid storage cavity, and then required reagents are pre-stored in the liquid storage cavity and the detection structure, so that the closed nucleic acid automatic extraction and amplification detection is realized, the risk of interface cross contamination is avoided, and the reliability of the detection result is also improved.
Meanwhile, according to the nucleic acid analysis card box provided by the embodiment of the invention, the liquid storage cavities and the auxiliary liquid storage cavities are distributed on two sides of the silica gel film transfer part 200 and are sequentially distributed along the moving direction of the silica gel film transfer part 200, so that the space in the set direction is fully utilized, the compactness of the whole nucleic acid analysis card box is improved, the occupied area of the whole nucleic acid analysis card box is effectively reduced, and the popularization of a molecular diagnosis technology is further promoted.
In the above-mentioned nucleic acid analysis card box, when pellosil 700 intercommunication stock solution chamber and the supplementary stock solution chamber rather than corresponding, the liquid in the stock solution intracavity can get into supplementary stock solution chamber. In order to facilitate the liquid in the liquid storage cavity to enter the auxiliary liquid storage cavity, the whole auxiliary liquid storage cavity can be selected to be positioned under the liquid storage cavity corresponding to the auxiliary liquid storage cavity, and the liquid in the liquid storage cavity flows into the auxiliary liquid storage cavity by the self gravity of the liquid. However, the silicone membrane can have certain effect of blockking up to liquid, and in order to improve the circulation, preferably when silicone membrane 700 intercommunication stock solution chamber and the auxiliary stock solution chamber rather than corresponding, the liquid in the stock solution intracavity can get into rather than the auxiliary stock solution intracavity that corresponds under the centrifugal action. At this time, when the nucleic acid analysis cartridge is centrifuged, the reservoir chamber is located near the proximal end of the nucleic acid analysis cartridge and the auxiliary reservoir chamber is located near the distal end of the nucleic acid analysis cartridge.
Because the liquid in the liquid storage cavity flows into the auxiliary liquid storage cavity under the centrifugal action, the liquid in the auxiliary liquid storage cavity can easily flow back to the liquid storage cavity in the centrifugal movement process. In order to avoid the above problems, the bottom end of the auxiliary liquid storage cavity is preferably lower than the bottom end of the liquid storage cavity corresponding to the auxiliary liquid storage cavity so as to limit the liquid in the auxiliary liquid storage cavity from flowing back to the liquid storage cavity corresponding to the auxiliary liquid storage cavity.
It is understood that the lower end of the auxiliary reservoir chamber is lower than the corresponding reservoir chamber, which means the state of the nucleic acid analysis cartridge when it is normally set, i.e., the state of the nucleic acid analysis cartridge when it is centrifuged.
The height difference between the bottom end of the auxiliary liquid storage cavity and the bottom end of the corresponding liquid storage cavity is selected according to actual needs, and the liquid in the auxiliary liquid storage cavity can not flow back to the liquid storage cavity, so that the liquid storage cavity is not limited in the embodiment.
The specific structures and shapes of the liquid storage cavity and the auxiliary liquid storage cavity are selected according to actual needs, and the embodiment does not limit the specific structures and shapes.
In the above-mentioned nucleic acid analysis cartridge, the liquid in the detection structure is detected by centrifugation, and for the convenience of detection, it is preferable that the detection structure is located on the bottom side of the auxiliary reservoir. Thus, the space in the vertical direction is utilized, and the structural compactness is improved.
It is understood that the detection structure is located at the bottom side of the auxiliary reservoirs, which means that the detection structure is located at the bottom side of all the auxiliary reservoirs, and is in a state where the nucleic acid analysis cartridge is normally set, i.e., in a state where the nucleic acid analysis cartridge is centrifuged.
The above-mentioned silicone membrane transfer member 200 needs to be driven by the transfer driving means 1300 to realize the movement in the set direction, i.e. the transfer driving means 1300 is used for driving the silicone membrane transfer member 200 to move in the set direction. In order to facilitate the driving of the transfer drive 1300, the silicone membrane transfer unit 200 is provided with a drive connection structure 201 that can be connected to the transfer drive 1300.
The specific structure of the driving connection structure 201 can be designed according to the structure of the selected transfer driving device 1300, which is not limited in this embodiment.
The silicone membrane 700 is fixed to the silicone membrane transfer unit 200, and the fixing structure of the silicone membrane 700 is selected according to actual needs. For convenience of installation, it is preferable that the above-mentioned silicone membrane transfer unit 200 is provided with a fixing groove 203 and a communication hole 202 communicating with the fixing groove 203, and the silicone membrane 700 is fixed in the fixing groove 203 by a fixing ring 800. In order to facilitate the liquid in the liquid storage chamber to flow into the auxiliary liquid storage chamber, it is preferable that the fixing ring 800 is adjacent to the auxiliary liquid storage chamber, and the communication hole 202 is adjacent to the liquid storage chamber.
It can be understood that, when the silicone film 700 communicates the corresponding reservoir and the auxiliary reservoir, the communication hole 202 communicates with the reservoir, and the fixing groove 203 communicates with the auxiliary reservoir.
And selecting the specific structure of the detection structure according to actual needs. For the convenience of detection, the detection structure preferably includes: the device comprises a sample feeding channel 309, an isolation valve 308, a detection cavity 310, a waste liquid cavity 311 and a ventilation structure, wherein the detection cavity 310, the waste liquid cavity 311 and the ventilation structure are all communicated with the sample feeding channel 309, the waste liquid cavity 311 is positioned at the downstream of the detection cavity 310, the ventilation structure is positioned at the downstream of the waste liquid cavity 311, and the sample feeding channel 309 is communicated with an auxiliary liquid storage cavity positioned at the last along a set direction; the isolation valve 308 is disposed on the sample channel 309 and can block and block the sample channel 309.
It will be appreciated that when the isolation valve 308 is closed, the isolation valve 308 disconnects the sample channel 309; when the isolation valve 308 is opened, the isolation valve 308 conducts the sample channel 309. In practice, the isolation valve 308 may be selected to open when the nucleic acid analysis cartridge is locally heated or centrifuged at a set rate.
One or more than two detection chambers 310 may be provided. When the number of the detection chambers 310 is two or more, the detection chambers 310 are sequentially distributed along the sample channel 309.
The specific number of the reservoir chambers and the detection chambers 310 can be set by those skilled in the art according to practical situations, and is not limited specifically herein.
Note that a desired reagent is stored in the reservoir chamber and the detection chamber in advance, for example, a detection reagent is immobilized in the detection chamber 310 in advance. The waste liquid chamber 311 is used to store excess reaction liquid.
In order to isolate the sample channel 309, a hydrophobic layer, such as a paraffin layer, is disposed in the isolation valve 308.
In order to facilitate ventilation and exhaust, the ventilation structure comprises: a ventilation channel 312 communicated with the sample feeding channel 309, a ventilation fixing hole 307 communicated with the ventilation channel 312, and a hydrophobic breathable film 900 sealed in the ventilation fixing hole 307; wherein the ventilation fixing hole 307 is provided on the cartridge body. It will be appreciated that the port 313 of the vent passage 312 communicates with the vent securement hole 307.
In practical applications, other structures may be selected to achieve ventilation and air exhaust, and are not limited to the above embodiments.
In the above-mentioned nucleic acid analysis cartridge, the specific structure of the cartridge body may be designed according to actual needs. Preferably, the cartridge body includes: the liquid storage box 100, the detection box 300, the first cover plate 500, the second cover plate 1000 and the third cover plate 1100.
The liquid storage cavity, the sample adding channel 104 and the air exhaust channel 105 are all arranged on the liquid storage box 100, and the first cover plate 500 is hermetically connected with the liquid storage box 100 to seal the liquid storage cavity, the sample adding channel 104 and the air exhaust channel 105; the auxiliary liquid storage cavity and the detection structure are arranged on the detection box 300, the second cover plate 1000 is connected with the detection box 300 in a sealing mode to form the detection structure, and the third cover plate 1100 is connected with the detection box 300 in a sealing mode to form the auxiliary liquid storage cavity.
The silicone membrane transfer member 200 is disposed between the liquid storage box 100 and the detection box 300, and both the liquid storage box 100 and the detection box 300 are hermetically connected to the silicone membrane transfer member 200.
It is understood that the silicone membrane transfer member 200 seals one port of the reservoir, and the first cover plate 500 seals the other port of the reservoir as well as the sample addition channel 104 and the air discharge channel 105; the silicone membrane transfer member 200 seals one port of the auxiliary reservoir and the third cover plate 1100 seals the other port of the auxiliary reservoir.
The first cover plate 500, the reservoir cartridge 100, the silicone film transfer member 200, the detection cartridge 300, and the third cover plate 1100 are sequentially disposed. When the detecting structure is located at the bottom side of the auxiliary liquid storage cavity, the second cover plate 1000 is located at the bottom side of the detecting box 300.
In order to facilitate the mounting of the silicone membrane transfer member 200, it is preferable that the above-described cartridge 300 is provided with a housing groove 301 for housing the silicone membrane transfer member 200.
Above-mentioned holding tank 301 holds pellosil transfer part 200, has also realized the location to pellosil transfer part 200 simultaneously, guarantees that pellosil transfer part 200 moves along setting for the direction.
The size and shape of the accommodating groove 301 are designed according to the size and shape of the silicone membrane transfer member 200, which is not limited in this embodiment.
In the practical application process, the accommodating groove 301 may be optionally formed in the liquid storage box 100, or the liquid storage box 100 and the detection box 300 are both provided with grooves, and the grooves on the liquid storage box 100 and the detection box 300 are spliced to form the accommodating cavity for accommodating the silicone membrane transfer component 200, which is not limited to the above embodiment.
The liquid storage box 100 and the detection box 300 are hermetically connected with the silicone membrane transfer part 200. The specific sealing structure is selected according to actual needs.
Preferably, the liquid storage cavity is provided with a first mounting groove 103, the auxiliary liquid storage cavity is provided with a second mounting groove 303, a sealing ring 600 is arranged in the first mounting groove 103 and the second mounting groove 303, and the liquid storage box 100 and the detection box 300 are both connected with the silicone membrane transfer part 200 in a sealing manner through the sealing ring 600.
The type of the sealing ring 600 is selected according to actual requirements, for example, the sealing ring 600 is made of an elastic polymer material, which is not limited in this embodiment.
Specifically, when liquid is stored in the auxiliary liquid storage chamber, the liquid level in the auxiliary liquid storage chamber is lower than the lower edge of the sealing ring 600.
In order to facilitate sealing, the bottom end of the auxiliary liquid storage cavity is lower than the bottom end of the liquid storage cavity corresponding to the auxiliary liquid storage cavity so as to limit liquid in the auxiliary liquid storage cavity to flow back to the liquid storage cavity corresponding to the auxiliary liquid storage cavity, and preferably, the port of the auxiliary liquid storage cavity close to the liquid storage cavity is smaller than the port of the auxiliary liquid storage cavity far away from the liquid storage cavity. Specifically, the cross section of the end, close to the liquid storage cavity, of the auxiliary liquid storage cavity is circular, and the cross section of the end, far away from the liquid storage cavity, of the auxiliary liquid storage cavity is non-circular. The specific shape of the non-circular shape is selected according to actual needs, and this embodiment does not limit this.
In order to facilitate installation, the liquid storage box 100 and the detection box 300 are fixedly connected through the fastener 400, the first cover plate 500 is hermetically connected with the liquid storage box 100 through bonding, and the second cover plate 1000 and the third cover plate 1100 are both hermetically connected with the detection box 300 through bonding.
One or more fasteners 400 may be provided. In order to improve the stability, it is preferable that the number of the fastening members 400 is two or more, and the fastening members 400 are distributed around the silicone membrane transfer member 200 to ensure the movement of the silicone membrane transfer member 200. To facilitate the installation of the second cover plate 1000, it is preferable that the fastening members 400 are distributed on the outer periphery of the second cover plate 1000.
Specifically, the liquid storage box 100 is provided with a first fixing hole 101 into which the fastening member 400 is inserted, and the detection box 300 is provided with a second fixing hole 304 into which the fastening member 400 is inserted.
In order to facilitate the understanding of the reaction result, the cartridge 300, the second cover 1000, and the third cover 1100 are transparent members. Preferably, the cartridge 300, the second cover 1000, and the third cover 1100 are all plastic transparencies.
Of course, other types of materials may be selected for the cartridge 300, the second cover 1000, and the third cover 1100, which are only exemplary and not limited in particular.
Further, the liquid storage box 100 and the first cover plate 500 are transparent members.
Based on the nucleic acid analysis cartridge provided in the above embodiment, an embodiment of the present invention also provides a nucleic acid analysis apparatus including the nucleic acid analysis cartridge described in the above embodiment.
Specifically, the above nucleic acid analysis apparatus further comprises: a carrier member 1200, a centrifuge 1400, a transfer drive 1300, and a detection device 1500.
The carrier 1200 carries the nucleic acid analysis cartridge, the centrifuge 1400 drives the carrier 1200 to rotate so as to make the nucleic acid analysis cartridge perform centrifugal motion, and the transfer drive 1300 is used for driving the silicone membrane transfer unit 200 to move along a set direction; the detecting device 1500 is used for detecting the reaction result of the detecting structure.
The specific structures and types of the carrying member 1200, the centrifugal device 1400, the transfer driving device 1300, and the detection device 1500 are selected according to actual needs, for example, the carrying member 1200 is a tray, the centrifugal device 1400 is a centrifugal motor, and the like, which is not limited in this embodiment.
In order to facilitate detection of the reaction result, the detecting device 1500 is an optical signal detecting device, and at this time, the portion of the cartridge body where the detecting structure is located is a transparent structure. The detecting device 1500 can detect the optical signal of the detecting structure and obtain the reaction result according to the optical signal.
Specifically, when the cartridge body includes the liquid storage cartridge 100, the detection cartridge 300, the first cover 500, the second cover 1000, and the third cover 1100, it is preferable that the detection cartridge 300, the second cover 1000, and the third cover 1100 are transparent members. The detecting device 1500 is used for detecting the optical signal of the detecting cavity 310 and obtaining the reaction result according to the optical signal.
In order to facilitate the reaction control, the above-mentioned carrying device 1200 is provided with a temperature control region for controlling the temperature of the detection chamber 310 of the detection structure. Specifically, the temperature control region is a temperature heating region, i.e., the temperature heating region is used for heating the detection chamber 310.
In the nucleic acid analysis apparatus, the number of nucleic acid analysis cartridges on the carrier member 1200 may be one, or two or more. In order to improve efficiency, it is preferable that the number of the nucleic acid analysis cartridges on the carriage 1200 is two or more, and the carriage 1200 operates all the nucleic acid analysis cartridges in synchronization. For the convenience of centrifugation, it is preferable that the above-described nucleic acid analysis cartridges are sequentially distributed in the rotation direction of the carrier member 1200.
The specific number and distribution of nucleic acid analysis cartridges, for example, the centrosymmetric arrangement, are selected according to the actual need, as long as the nucleic acid analysis cartridges are allowed to perform centrifugal motion.
Since the nucleic acid analysis cartridge provided in the above embodiment has the above technical effects, and the nucleic acid analysis device provided in the above embodiment has the above nucleic acid analysis cartridge, the above nucleic acid analysis device also has corresponding technical effects, and details are not repeated herein.
The nucleic acid analysis equipment provided by the embodiment of the invention can be used for automatically detecting and analyzing nucleic acid of a biological sample, namely, can be used for fully integrating automatically processing the sample and detecting and analyzing, has the advantages of simple operation process, high detection flux, higher clinical application value, full integration, higher detection stability and efficiency and lower cost, and promotes the popularization of molecular diagnosis.
In practical application, the method for using the nucleic acid analysis device mainly comprises the following steps:
s01: adding a sample from the sample addition channel 104 into the liquid storage cavity, and then placing the nucleic acid analysis card box on the bearing part 1200;
s02: the liquid transfer is completed through the cooperative control of the transfer driving device 1300 and the centrifugal device 1400, and the temperature control area of the bearing part 1200 is controlled according to the target temperature of the detection reaction, so as to realize the required biological detection reaction;
s03: the detection device 1500 reads the reaction result of the detection structure of the nucleic acid analysis cartridge.
The reservoir chamber and the detection structure are preliminarily stored with reagents required for the reaction.
The shape of the nucleic acid analysis cartridge is mainly determined by the shape of the silicone membrane transfer member 200, and the shapes of the silicone membrane transfer member 200 and the nucleic acid analysis cartridge are selected according to actual needs. The shape and the moving mode of the silica gel transfer member 200 are selected according to actual needs.
Two specific examples are provided below to further explain the nucleic acid analysis cartridge and the nucleic acid analysis apparatus provided in this example, according to the shape of the silicone membrane transfer member 200.
Implementing one step:
as shown in fig. 1 to 22, in the nucleic acid analysis cartridge according to the first embodiment, the silicone membrane transfer member 200 is in a strip shape, and the silicone membrane transfer member 200 linearly reciprocates in a predetermined direction, which is the longitudinal direction of the silicone membrane transfer member 200.
It is understood that the silicone membrane transfer unit 200 is linearly moved back and forth in a predetermined direction. In the reciprocating process of the silicone membrane transfer member 200, the silicone membrane transfer member 200 is easy to separate from the cartridge body, and in order to avoid the above problem, it is preferable that the silicone membrane transfer member 200 is provided with a limiting structure 204, and the limiting structure 204 is used for limiting the silicone membrane transfer member 200 to separate from the cartridge body.
Specifically, the stopper structure 204 restricts the separation of the silicone film transfer member 200 from the cartridge body by abutting against the cartridge body. In practical application, when the limiting structure 204 abuts against the cartridge body, preferably all the liquid storage cavities are located between the silicone membrane 700 and the limiting structure 204. Thus, the operation is facilitated.
The specific structure of the limiting structure 204 is designed according to actual needs, for example, the limiting structure 204 is a limiting protrusion or a limiting plate.
For the convenience of limiting and driving, the limiting structure 204 is disposed at one end of the silicone membrane transfer unit 200, and the other end of the silicone membrane transfer unit 200 is used for connecting with the transfer driving device 1300.
In the first embodiment, the cartridge body includes a liquid storage case 100, a detection case 300, a first cover 500, a second cover 1000, and a third cover 1100. For the specific structure of the entire cartridge body, please refer to the above description, which is not repeated herein.
The shapes of the above components are selected according to actual needs. Preferably, the liquid storage box 100, the first cover plate 500, the second cover plate 1000 and the third cover plate 1100 are all in a strip shape. Since the cartridge 300 needs to be provided with a detection structure, the shape of the cartridge 300 is preferably irregular.
In the first embodiment, in order to facilitate nucleic acid extraction and nucleic acid amplification detection, the cartridge preferably has three reservoirs, namely, a first reservoir 106a, a second reservoir 106b, and a third reservoir 106 c. The third reservoir 106c is the last reservoir, and the sample channel 309 is connected to the third reservoir 106 c. Correspondingly, the number of the auxiliary liquid storage cavities is also three, namely a first auxiliary liquid storage cavity 305a, a second auxiliary liquid storage cavity 305b and a third auxiliary liquid storage cavity 305c, the first liquid storage cavity 106a corresponds to the first auxiliary liquid storage cavity 305a, the second liquid storage cavity 106b corresponds to the second auxiliary liquid storage cavity 305b, and the third liquid storage cavity 106c corresponds to the third auxiliary liquid storage cavity 305 c.
In the above-mentioned nucleic acid analysis cartridge, when limit structure 204 of pellosil transfer unit 200 and stock solution box 100 and detection box 300 butt, pellosil 700 is located the one side that second stock solution chamber 106b was kept away from to first stock solution chamber 106a, and first stock solution chamber 106a, second stock solution chamber 106b and third stock solution chamber 106c do not all communicate with pellosil 700 on pellosil transfer unit 200, and first stock solution chamber 106a, second stock solution chamber 106b and third stock solution chamber 106c all are in encapsulated situation promptly.
In the cartridge for nucleic acid analysis provided in the first embodiment, the liquid storage cartridge 100 and the detection cartridge 300 are fixedly connected by fasteners 400, specifically, four fasteners 400 are provided and distributed at four corners of the liquid storage cartridge 100; the number of the first fixing holes 101 of the liquid storage box 100 and the number of the second fixing holes 304 of the detection box 300 are four. To avoid affecting the seal, the fasteners 400 are located at the periphery of the third cover plate 1100.
In the first embodiment, which provides the nucleic acid analysis cartridge according to the first embodiment, there is provided a nucleic acid analysis apparatus in which at least two nucleic acid analysis cartridges are sequentially arranged in the rotation direction of the carrier member 1200, as shown in fig. 22. In order to secure the reaction result, it is preferable that the nucleic acid analysis cartridges are uniformly distributed in the rotation direction of the carrier member 1200.
For convenience of installation and reduction of volume, the bearing member 1200 is preferably circular. At this time, the carrier member 1200 rotates around its axis, i.e., the nucleic acid analysis cartridges are sequentially distributed along the circumferential direction of the carrier member 1200.
In order to simplify the structure, the silicone membrane transfer sections 200 of at least two nucleic acid analysis cartridges are driven by the same transfer drive device 1300. The specific configuration of the transfer drive device 1300 is selected according to actual needs. Preferably, the transfer drive device 1300 includes: a driving jig 1301, a driving member for driving the driving jig 1301; wherein the silicone membrane transfer members 200 of at least two nucleic acid analysis cartridges are each fixed on the driving jig 1301.
For maximum simplicity of construction, it is preferred that all of the silicone membrane transfer members 200 on the carrier member 1200 are driven by the same transfer drive 1300.
In order to facilitate driving the silicone membrane transfer member 200 to move linearly in practical use, it is preferable that the transfer driving means 1300 is located on the top side of the silicone membrane transfer member 200.
The procedure for detecting nucleic acid using the nucleic acid analysis apparatus provided in the first embodiment is as follows:
(1) adding a sample to be detected into the first liquid storage cavity 106a through the sample adding channel 104, and driving the silicone membrane transfer part 200 to move to the position shown in fig. 11 by the transfer driving device 1300, at this time, the first liquid storage cavity 106a, the second liquid storage cavity 106b and the third liquid storage cavity 106c are all located between the silicone membrane 700 and the limiting structure 204, so that the first liquid storage cavity 106a, the second liquid storage cavity 106b and the third liquid storage cavity 106c are all in a sealed state, as shown in fig. 12, the centrifugal device 1400 drives the bearing part 1200 to rotate, so that the nucleic acid analysis card box rotates and vibrates, and the sample to be detected in the first liquid storage cavity 106a is fully cracked;
(2) after completing the sample lysis, the centrifugal device 1400 stops driving, the transfer driving device 1300 drives the silicone membrane transfer component 200 to move downward, so that the silicone membrane 700 is aligned with the first inlet 102a of the first reservoir 106a and the auxiliary first inlet 302a of the first auxiliary reservoir 305a, at this time, the silicone membrane 700 communicates the first reservoir 106a and the first auxiliary reservoir 305a, and then the centrifugal device 1400 drives the bearing component 1200 to rotate, so that the nucleic acid analysis cartridge is centrifuged, so that the solution in the first reservoir 106a enters the first auxiliary reservoir 305a through the silicone membrane 700, and the silicone membrane 700 captures the nucleic acid in the sample lysate, as shown in fig. 13 and 14;
(3) after the nucleic acid is captured by the silicone membrane 700, the centrifugal device 1400 stops driving, the transfer driving device 1300 drives the silicone membrane transfer component 200 to move downwards continuously, so that the silicone membrane 700 is aligned with the second inlet 102b of the second reservoir 106b and the auxiliary second inlet 302b of the second auxiliary reservoir 305b, that is, the silicone membrane 700 is communicated with the second reservoir 106b and the second auxiliary reservoir 305b, and then the centrifugal device 1400 drives the carrying component 1200 to rotate, so that the nucleic acid analysis cartridge is centrifuged, so that the cleaning solution in the second reservoir 106b enters the second auxiliary reservoir 305b through the silicone membrane 700, and the cleaning solution sufficiently cleans the silicone membrane 700, as shown in fig. 15 and 16;
(4) after the washing of the silicone membrane 700 is completed, the centrifugal device 1400 stops driving, the transfer driving device 1300 drives the silicone membrane transfer part 200 to move downwards continuously, so that the silicone membrane 700 is aligned with the third inlet 102c of the third liquid storage cavity 106c and the auxiliary third inlet 302c of the third auxiliary liquid storage cavity 305c, that is, the silicone membrane 700 is communicated with the third liquid storage cavity 106c and the third auxiliary liquid storage cavity 305c, then the centrifugal device 1400 drives the bearing part 1200 to rotate, so that the nucleic acid analysis cartridge is centrifuged, so that the nucleic acid eluent in the third liquid storage cavity 106c enters the third auxiliary liquid storage cavity 305c through the silicone membrane 700, and the nucleic acid bound on the silicone membrane 700 is sufficiently eluted by the eluent, as shown in fig. 17 and fig. 18;
(5) after the nucleic acid elution is completed, the centrifugal device 1400 stops driving, and the transfer driving device 1300 drives the silicone membrane transfer component 200 to continue to move downwards, so that the first liquid storage cavity 106a, the second liquid storage cavity 106b and the third liquid storage cavity 106c are all in a sealed state, that is, the silicone membrane 700 is far away from the first liquid storage cavity 106a, the second liquid storage cavity 106b and the third liquid storage cavity 106c, as shown in fig. 19 and fig. 20;
(6) after the sealing is completed, the centrifugal device 1400 drives the bearing part 1200 to rotate, so that the nucleic acid eluent in the third auxiliary liquid storage cavity 305c enters a downstream detection structure through the through hole 306, the nucleic acid eluent breaks through the isolation valve 308 through high-speed centrifugation and is distributed into the plurality of detection cavities 310 through the sample injection pipeline 309, as shown in fig. 21, the waste liquid cavity 311 contains redundant solution, the vent channel 312 is communicated with the vent fixing hole 307, and the hydrophobic gas-permeable membrane 900 seals the vent fixing hole 307, so that the nucleic acid analysis cartridge can be sealed in the whole process, namely the nucleic acid analysis cartridge is not communicated with the outside, and the pollution is avoided; the temperature control region of the bearing member 1200 controls the temperature of the detection chamber 310, so as to amplify the nucleic acid in the detection chamber 310 by variable temperature amplification or constant temperature amplification, and the detection device 1500 scans and detects the signal value of the detection chamber 310 in real time to obtain the detection result.
In the step (1), the nucleic acid analysis cartridge is in the first state; in the above step (2), the nucleic acid analysis cartridge is in the second state; in the above step (3), the nucleic acid analysis cartridge is in the third state; in the above step (4), the nucleic acid analysis cartridge is in the fourth state; in the above step (5), the nucleic acid analysis cartridge is in a fifth state; in the above step (6), the nucleic acid analysis cartridge is in the sixth state.
Example two
As shown in fig. 23 to 44, in the cartridge for nucleic acid analysis according to the second embodiment, the silicone membrane transfer member 200 is circular, and the silicone membrane transfer member 200 is rotated around its axis and set in the circumferential direction of the silicone membrane transfer member 200.
In order to facilitate the rotation of the silicone membrane transfer member 200, the driving connection structure 201 is preferably disposed on one side of the silicone membrane transfer member 200, and the driving connection structure 201 is coaxial with the silicone membrane transfer member 200. For the convenience of installation, the driving connection structure 201 is a connection column, and the cross section of the connection column may be a circle or a regular polygon.
In the second embodiment, the cartridge body includes: the liquid storage box 100, the detection box 300, the first cover plate 500, the second cover plate 1000 and the third cover plate 1100. For the specific structure of the entire cartridge body, please refer to the above description, which is not repeated herein.
For the convenience of setting up stock solution chamber and supplementary stock solution chamber along the circumference of pellosil transfer unit 200, above-mentioned stock solution box 100 and detection box 300 all have the connecting plate and protrusion in the connecting plate and be columniform bulge. Wherein, the liquid storage cavity is arranged on the convex part of the liquid storage box 100, and the auxiliary liquid storage cavity is arranged on the convex part of the detection box 300. At this time, it is preferable that the first and third cover plates 500 and 1100 be circular plates and the second cover plate 1000 be a rectangular plate.
The above-mentioned detection box 300 is provided with holding tank 301, and this holding tank 301 is circular, and 301 in the holding tank is embedded completely to pellosil transfer part 200, need not to set up limit structure 204.
Based on the nucleic acid analysis cartridge provided in the second embodiment, the second embodiment further provides a nucleic acid analysis apparatus, as shown in fig. 44, in which the silicone membrane transfer member 200 is circular, and at least two nucleic acid analysis cartridges on the carrier member 1200 are sequentially distributed along the rotation direction of the carrier member 1200.
In order to facilitate the rotation of each of the silica gel membrane transfer units 200, it is preferable that the transfer driving means 1300 be in one-to-one correspondence with the nucleic acid analysis cartridges. Further, the above-described transfer driving device 1300 is located at one end of the nucleic acid analysis cartridge near the rotation axis of the carrier member 1200.
In practical use, it is preferable that the carrier member 1200 is rectangular, and the number of the nucleic acid analysis cartridges is at least two, and is centrosymmetric with respect to the rotational axis of the carrier member 1200.
The procedure for detecting nucleic acid using the nucleic acid analysis apparatus provided in the second embodiment is as follows:
(1) adding a sample to be detected into the first liquid storage cavity 106a through the sample adding channel 104, driving the silicone membrane transfer member 200 to rotate to the position shown in fig. 33 by the transfer driving device 1300, at this time, the first liquid storage cavity 106a, the second liquid storage cavity 106b and the third liquid storage cavity 106c are all in a sealed state, as shown in fig. 34, the centrifugal device 1400 drives the bearing member 1200 to rotate, so that the nucleic acid analysis card box rotates and vibrates, and the sample to be detected in the first liquid storage cavity 106a is fully cracked and the silicone membrane is combined with nucleic acid;
(2) after completing the sample lysis, the centrifugal device 1400 stops driving, the transfer driving device 1300 drives the silicone membrane transfer component 200 to rotate counterclockwise, so that the silicone membrane 700 aligns with the first inlet 102a of the first liquid storage cavity 106a and the auxiliary first inlet 302a of the first auxiliary liquid storage cavity 305a, at this time, the silicone membrane 700 communicates the first liquid storage cavity 106a and the first auxiliary liquid storage cavity 305a, then the centrifugal device 1400 drives the bearing component 1200 to rotate, so that the solution in the first liquid storage cavity 106a enters the first auxiliary liquid storage cavity 305a through the silicone membrane 700, and the silicone membrane 700 captures the nucleic acid in the sample lysate, as shown in fig. 35 and 36;
(3) after the nucleic acid is captured by the silicone membrane 700, the centrifugal device 1400 stops driving, the transfer driving device 1300 drives the silicone membrane transfer component 200 to continue to rotate counterclockwise, so that the silicone membrane 700 is aligned with the second inlet 102b of the second reservoir 106b and the auxiliary second inlet 302b of the second auxiliary reservoir 305b, that is, the silicone membrane 700 is communicated with the second reservoir 106b and the second auxiliary reservoir 305b, and then the centrifugal device 1400 drives the carrying component 1200 to rotate, so that the cleaning solution in the second reservoir 106b enters the second auxiliary reservoir 305b through the silicone membrane 700, and the cleaning solution sufficiently cleans the silicone membrane 700, as shown in fig. 37 and 38;
(4) after the washing of the silicone membrane 700 is completed, the centrifugal device 1400 stops driving, the transfer driving device 1300 drives the silicone membrane transfer component 200 to continue to rotate counterclockwise, so that the silicone membrane 700 is aligned with the third inlet 102c of the third liquid storage cavity 106c and the auxiliary third inlet 302c of the third auxiliary liquid storage cavity 305c, that is, the silicone membrane 700 is communicated with the third liquid storage cavity 106c and the third auxiliary liquid storage cavity 305c, then the centrifugal device 1400 drives the bearing component 1200 to rotate, so that the eluent in the third liquid storage cavity 106c enters the third auxiliary liquid storage cavity 305c through the silicone membrane 700, and the eluent is fully combined with the nucleic acid on the silicone membrane 700, as shown in fig. 39 and fig. 40;
(5) after the nucleic acid elution is completed, the centrifugal device 1400 stops driving, and the transfer driving device 1300 drives the silicone membrane transfer component 200 to continue to rotate counterclockwise, so that the first liquid storage cavity 106a, the second liquid storage cavity 106b and the third liquid storage cavity 106c are all in a sealed state, that is, the silicone membrane 700 is far away from the first liquid storage cavity 106a, the second liquid storage cavity 106b and the third liquid storage cavity 106c, as shown in fig. 41 and 42;
(6) after sealing, the centrifugal device 1400 drives the bearing part 1200 to rotate, so that the nucleic acid eluent in the third auxiliary liquid storage cavity 305c enters the detection structure through the through hole 306, specifically, through high-speed centrifugation, the nucleic acid eluent breaks through the isolation valve 308 and is distributed into a plurality of detection cavities 310 through the sample introduction pipeline 309, as shown in FIG. 43, the waste liquid chamber 311 contains the excess solution, the vent channel 312 is connected to the vent fixing hole 307, and the hydrophobic gas-permeable membrane 900 seals the vent fixing hole 307, so that the whole process of the nucleic acid analysis cartridge can be sealed, that is, the nucleic acid analysis cartridge is not communicated with the outside, so that contamination is prevented, the temperature control region of the carrier 1200 controls the temperature of the detection chamber 310, so that amplification of nucleic acid in the detection chamber 310 is realized, the temperature-variable amplification or the constant-temperature amplification can be adopted, and the detection device 1500 scans and detects the signal value of the detection cavity 310 in real time to obtain a detection result.
In the step (1), the nucleic acid analysis cartridge is in the first state; in the above step (2), the nucleic acid analysis cartridge is in the second state; in the above step (3), the nucleic acid analysis cartridge is in the third state; in the above step (4), the nucleic acid analysis cartridge is in the fourth state; in the above step (5), the nucleic acid analysis cartridge is in a fifth state; in the above step (6), the nucleic acid analysis cartridge is in the sixth state.
In the description of the present invention, it is to be understood that the terms "upper", "lower", "top", "bottom", "inner", "outer", and the like, indicate orientations or positional relationships based on those shown in the drawings, and are used only for convenience in describing the present invention and for simplicity in description, and do not indicate or imply that the referenced devices or elements must have a particular orientation, be constructed and operated in a particular orientation, and thus, are not to be construed as limiting the present invention.
The previous description of the disclosed embodiments is provided to enable any person skilled in the art to make or use the present invention. Various modifications to these embodiments will be readily apparent to those skilled in the art, and the generic principles defined herein may be applied to other embodiments without departing from the spirit or scope of the invention. Thus, the present invention is not intended to be limited to the embodiments shown herein but is to be accorded the widest scope consistent with the principles and novel features disclosed herein.

Claims (12)

1. A nucleic acid analysis cartridge, comprising: the cartridge comprises a cartridge body and a silicone membrane transfer component (200) which is arranged on the cartridge body and can move along a set direction;
the card box body is provided with at least two liquid storage cavities, at least two auxiliary liquid storage cavities, a detection structure, a sample adding channel (104) and an exhaust channel (105), wherein the sample adding channel and the exhaust channel are communicated with the liquid storage cavities, the sample adding channels and the exhaust channels are sequentially distributed along the set direction, the auxiliary liquid storage cavities correspond to the liquid storage cavities one by one, and the auxiliary liquid storage cavity positioned at the last along the set direction can be communicated with the detection structure;
the liquid storage cavity, the sample adding channel (104) and the air exhaust channel (105) are all positioned on one side of the silicone membrane transfer component (200), and the auxiliary liquid storage cavity and the detection structure are all positioned on the other side of the silicone membrane transfer component (200);
stock solution chamber place face with supplementary stock solution chamber place face all with pellosil transfer part (200) sealing connection, pellosil transfer part (200) are fixed with pellosil (700), pellosil transfer part (200) are followed it can make to set for the direction removal pellosil (700) communicate every in proper order stock solution chamber and rather than corresponding supplementary stock solution chamber.
2. The nucleic acid analysis cartridge according to claim 1, wherein when the silicone membrane (700) is in communication with the reservoir chamber and the auxiliary reservoir chamber corresponding thereto, liquid in the reservoir chamber can enter the auxiliary reservoir chamber corresponding thereto under centrifugation;
the bottom end of the auxiliary liquid storage cavity is lower than the bottom end of the liquid storage cavity corresponding to the auxiliary liquid storage cavity so as to limit liquid in the auxiliary liquid storage cavity to flow back to the liquid storage cavity corresponding to the auxiliary liquid storage cavity.
3. The nucleic acid analysis cartridge of claim 2, wherein the detection structure is located on a bottom side of the auxiliary reservoir.
4. The nucleic acid analysis cartridge of claim 1,
the silicone film transfer part (200) is in a strip shape, the silicone film transfer part (200) moves linearly along the set direction, and the set direction is the length direction of the silicone film transfer part (200); the silicone film transfer component (200) is provided with a limiting structure (204), and the limiting structure (204) is used for limiting the silicone film transfer component (200) to be separated from the card box body;
or the silicone membrane transfer part (200) is circular, the silicone membrane transfer part (200) rotates around the axis of the silicone membrane transfer part, and the set direction is the circumferential direction of the silicone membrane transfer part (200).
5. The nucleic acid analysis cartridge according to claim 1, wherein the silicone membrane transfer member (200) is provided with a drive connection structure (201) connectable to a transfer drive means (1300), the transfer drive means (1300) being for driving the silicone membrane transfer member (200) to move in the set direction;
the silicone membrane transfer component (200) is provided with a fixing groove (203) and a communication hole (202) communicated with the fixing groove (203), and the silicone membrane (700) is fixed in the fixing groove (203) through a fixing ring (800).
6. The nucleic acid analysis cartridge of claim 1, wherein the detection structure comprises: the device comprises a sample feeding channel (309), an isolation valve (308), a detection cavity (310), a waste liquid cavity (311) and a ventilation structure;
wherein the detection cavity (310), the waste liquid cavity (311) and the ventilation structure are all communicated with the sample feeding channel (309), the waste liquid cavity (311) is positioned at the downstream of the detection cavity (310), the ventilation structure is positioned at the downstream of the waste liquid cavity (311), and the sample feeding channel (309) is communicated with the auxiliary liquid storage cavity positioned at the last along the set direction; the isolation valve (308) is arranged on the sample feeding channel (309), and the isolation valve (308) can cut off and conduct the sample feeding channel (309).
7. The nucleic acid analysis cartridge of claim 6, wherein the vent structure comprises: a vent channel (312) communicated with the sample feeding channel (309), a vent fixing hole (307) communicated with the vent channel (312), and a hydrophobic breathable membrane (900) sealed in the vent fixing hole (307); wherein the vent fixing hole (307) is provided on the cartridge body.
8. The nucleic acid analysis cartridge of any one of claims 1-7, wherein the cartridge body comprises: the liquid storage box (100), the detection box (300), the first cover plate (500), the second cover plate (1000) and the third cover plate (1100);
the liquid storage cavity, the sample adding channel (104) and the air exhaust channel (105) are all arranged on the liquid storage box (100), and the first cover plate (500) is hermetically connected with the liquid storage box (100) to seal the liquid storage cavity, the sample adding channel (104) and the air exhaust channel (105);
the auxiliary liquid storage cavity and the detection structure are arranged on the detection box (300), the second cover plate (1000) is in sealing connection with the detection box (300) to seal the detection structure, and the third cover plate (1100) is in sealing connection with the detection box (300) to seal the auxiliary liquid storage cavity;
the silicone membrane transfer part (200) is arranged between the liquid storage box (100) and the detection box (300), and the liquid storage box (100) and the detection box (300) are hermetically connected with the silicone membrane transfer part (200);
the detection box (300) is provided with an accommodating groove (301) for accommodating the silicone membrane transfer part (200).
9. The nucleic acid analysis cartridge of claim 8,
the liquid storage cavity is provided with a first mounting groove (103), the auxiliary liquid storage cavity is provided with a second mounting groove (303), sealing rings (600) are arranged in the first mounting groove (103) and the second mounting groove (303), and the liquid storage box (100) and the detection box (300) are both connected with the silicone film transfer component (200) in a sealing manner through the sealing rings (600);
stock solution box (100) with detect box (300) through fastener (400) fixed connection, first apron (500) through bonding with stock solution box (100) sealing connection, second apron (1000) with third apron (1100) all through bonding with detect box (300) sealing connection.
10. The nucleic acid analysis cartridge according to claim 8, wherein the cartridge (300), the second cover plate (1000) and the third cover plate (1100) are all transparent members.
11. A nucleic acid analysis apparatus comprising a nucleic acid analysis cartridge, wherein the nucleic acid analysis cartridge is the nucleic acid analysis cartridge according to any one of claims 1 to 10;
the nucleic acid analysis apparatus further includes: a carrier member (1200), a centrifuge device (1400), a transfer drive device (1300), and a detection device (1500);
wherein the carrying part (1200) carries the nucleic acid analysis cartridge, the centrifugal device (1400) drives the carrying part (1200) to rotate so as to enable the nucleic acid analysis cartridge to do centrifugal motion, and the transfer driving device (1300) is used for driving the silica gel membrane transfer part (200) to move along the set direction; the detection device (1500) is used for detecting a reaction result of the detection structure.
12. The nucleic acid analysis apparatus according to claim 11, wherein the nucleic acid analysis cartridges are at least two and are sequentially arranged in the rotational direction of the carrier member (1200);
if the silica gel film transfer part (200) is in a strip shape, the silica gel film transfer parts (200) of at least two nucleic acid analysis card boxes are driven by the same transfer driving device (1300); if the silica gel membrane transfer part (200) is circular, the transfer driving devices (1300) correspond to the nucleic acid analysis card boxes one by one.
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