CN114931986A - Nucleic acid detection micro-fluidic device and nucleic acid detection method thereof - Google Patents
Nucleic acid detection micro-fluidic device and nucleic acid detection method thereof Download PDFInfo
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- CN114931986A CN114931986A CN202210365691.XA CN202210365691A CN114931986A CN 114931986 A CN114931986 A CN 114931986A CN 202210365691 A CN202210365691 A CN 202210365691A CN 114931986 A CN114931986 A CN 114931986A
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- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
- B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
- B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
- B01L3/5027—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6806—Preparing nucleic acids for analysis, e.g. for polymerase chain reaction [PCR] assay
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- 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
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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
- C12Q1/70—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving virus or bacteriophage
- C12Q1/701—Specific hybridization probes
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- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2200/00—Solutions for specific problems relating to chemical or physical laboratory apparatus
- B01L2200/06—Fluid handling related problems
- B01L2200/0689—Sealing
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2200/00—Solutions for specific problems relating to chemical or physical laboratory apparatus
- B01L2200/10—Integrating sample preparation and analysis in single entity, e.g. lab-on-a-chip concept
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
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- B01L2300/00—Additional constructional details
- B01L2300/08—Geometry, shape and general structure
- B01L2300/0861—Configuration of multiple channels and/or chambers in a single devices
- B01L2300/0867—Multiple inlets and one sample wells, e.g. mixing, dilution
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2400/00—Moving or stopping fluids
- B01L2400/04—Moving fluids with specific forces or mechanical means
- B01L2400/0475—Moving fluids with specific forces or mechanical means specific mechanical means and fluid pressure
- B01L2400/0478—Moving fluids with specific forces or mechanical means specific mechanical means and fluid pressure pistons
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2400/00—Moving or stopping fluids
- B01L2400/04—Moving fluids with specific forces or mechanical means
- B01L2400/0475—Moving fluids with specific forces or mechanical means specific mechanical means and fluid pressure
- B01L2400/0481—Moving fluids with specific forces or mechanical means specific mechanical means and fluid pressure squeezing of channels or chambers
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2400/00—Moving or stopping fluids
- B01L2400/04—Moving fluids with specific forces or mechanical means
- B01L2400/0475—Moving fluids with specific forces or mechanical means specific mechanical means and fluid pressure
- B01L2400/0487—Moving fluids with specific forces or mechanical means specific mechanical means and fluid pressure fluid pressure, pneumatics
- B01L2400/049—Moving fluids with specific forces or mechanical means specific mechanical means and fluid pressure fluid pressure, pneumatics vacuum
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Abstract
The invention relates to the field of nucleic acid detection, in particular to a nucleic acid detection microfluidic device and a nucleic acid detection method thereof. The test paper box comprises a housin, the inside piston cavity that is equipped with of casing, piston rod under the piston cavity internal piston connection down, the upper portion of piston cavity communicates first reagent cavity and second reagent cavity respectively down, the lower part intercommunication reagent mixing component of piston cavity down, one side and the test paper strip mounting groove intercommunication of reagent mixing component, insert the chromatography test paper strip in the test paper strip mounting groove, the chromatography test paper strip extends into inside the reagent mixing component, casing one side is equipped with the opening, the opening is equipped with press device, press device extends into inside and through gear drive connection of casing piston rod down. The invention is portable and easy to operate, has quick detection time, high detection result accuracy and high biological safety.
Description
Technical Field
The invention relates to the field of nucleic acid detection, in particular to a nucleic acid detection microfluidic device and a nucleic acid detection method thereof.
Background
In the traditional nucleic acid detection method, after a nasopharyngeal swab sample is collected, a special operator needs to perform sample inactivation, lysis, magnetic bead enrichment extraction and final PCR amplification detection on the collected sample in a laboratory with more than two levels of biosafety. The traditional nucleic acid detection method has high requirement on environment, complex operation, expensive equipment and long time consumption, can not carry out on-site rapid detection, and greatly limits the application and popularization of the method.
A reverse transcription enhanced recombinase-dependent isothermal nucleic acid amplification technology combined with a Lateral flow chromatography test strip is a technology for directly detecting RT-eRDA amplification products by combining a reverse transcription enhanced recombinase-dependent isothermal nucleic acid amplification technology (RT-eRDA technology) with a Lateral flow chromatography test strip (LFD) technology. The RT-eRDA-LFD technology is based on the RT-eRDA technology, and mainly utilizes a specific probe (nfo probe) with a 5 'end labeled with a fluorescent group (usually FAM) and a reverse primer with a 5' end labeled with biotin to incubate at constant temperature of 37-42 ℃ for 20-40 minutes to form a double-labeled amplification product. When the test strip is used for detection, an amplification product to be detected flows to a binding region which is coated with FAM antibody-nano gold particles in advance by utilizing the chromatography. The FAM antibody-gold nanoparticles form immune complexes after binding with FAM in the double-labeled amplification product. The detection line (Test, T) is coated with biotin antibody, and when the immune complex diffuses to the detection line, biotin in the double-labeled amplification product is captured by the biotin antibody and forms a macroscopic immune complex. The quality Control line (Control, C) is coated with a secondary antibody, can be combined with FAM antibody-nano gold particles to generate a color reaction and indicate a detection result.
Compared with the traditional nucleic acid detection method, the method has the characteristics of quick response, visual judgment of detection results, no need of special equipment and the like, and is particularly suitable for application scenes such as low-resource areas, POCT, on-site quick detection and the like. However, the method still has the problems of complicated operation steps, poor biological safety, low sensitivity, low integration degree and the like.
Chinese patent CN 111733288A discloses a nucleic acid detection method and device and application in COVID-19 detection, firstly adopts RT-RAA-LFD technology to establish a micro-fluidic chip for rapidly detecting new coronavirus, and can be used for clinical field detection through specificity, sensitivity and actual sample analysis, thereby providing a sensitive and reliable new method for the instant detection of new coronavirus. The device's inner structure is fixed slot, and mutual UNICOM, and this leads to its can't realize pre-filling and isolation of material, like buffer solution, freeze-drying reagent etc. so all application of sample operations all rely on the user to accomplish, and is comparatively loaded down with trivial details. And because its direction of placing that relies on the device realizes detecting the flow, lacks the mistake proofing mechanism of preventing staying, in case unexpected maloperation of user, such as situation such as turn over in stage or application of sample in-process will lead to detecting inefficacy, and the kit is scrapped. Based on the above problems, the invention is more biased to experimental conceptual devices rather than industrial positioning.
Chinese patent CN 112760193A discloses a nucleic acid extraction/detection device, which is characterized in that lysate, cleaning solution, eluent and nucleic acid amplification solution are pre-embedded in different closed and isolated areas of the device, and then the device is simply rotated and extruded, so that related liquid sequentially passes through a nucleic acid adsorption area, sample cracking, nucleic acid purification and nucleic acid amplification detection can be gradually realized, and the requirements of sample inlet-result outlet in rapid detection of nucleic acid on site are really realized. Meanwhile, the device is a disposable closed device, so that the problem of cross contamination of amplification products can be effectively reduced and avoided. However, the mode of puncturing the sealing film by the ejector pin is mainly adopted to realize the flowing and mixing of all materials, and in actual industrial experience, the problem that the diameter of a channel is random after puncturing and the consistency is poor generally exists in the mode of puncturing the sealing film, so that the liquid mixing proportion cannot be reliably controlled, and the reaction effect and the detection result are influenced.
Therefore, there is an urgent need to develop a disposable RT-edra-LFD nucleic acid detection device that integrates nucleic acid extraction, amplification, and detection, has a high degree of integration, is inexpensive, portable, convenient to operate, and has high biosafety, and can perform closed-loop processing, detection, and recovery treatment of a nucleic acid sample on site by a simple trigger mechanism, thereby getting rid of dependence on biosafety laboratories. There are very few devices currently available on the domestic market that can achieve the above-described effects.
Disclosure of Invention
Aiming at the problems in the prior art, the invention provides the nucleic acid detection microfluidic device which is portable, easy to operate, used for nucleic acid detection, quick in detection time, high in detection result accuracy and high in biological safety, and further provides the nucleic acid detection method adopting the nucleic acid detection microfluidic device.
The micro-fluidic device for nucleic acid detection comprises a shell, wherein a lower piston cavity is arranged in the shell, a piston in the lower piston cavity is connected with a lower piston rod, the upper part of the lower piston cavity is respectively communicated with a first reagent cavity and a second reagent cavity, the lower part of the lower piston cavity is communicated with a reagent mixing component, one side of the reagent mixing component is communicated with a test strip mounting groove, a chromatography test strip is inserted into the test strip mounting groove and extends into the reagent mixing component, an opening is arranged on one side of the shell, a pressing device is arranged on the opening, and the pressing device extends into the shell and is connected with the lower piston rod through gear transmission.
Preferably, the top of the first reagent cavity is communicated with an upper piston cavity, a piston rod is connected to the piston in the upper piston cavity, the upper piston cavity is communicated with a first air guide buffer cavity through a first air guide pipe, the first air guide buffer cavity is communicated with a second air guide buffer cavity, the second air guide buffer cavity is communicated with one side of a lower piston cavity, the top of the second reagent cavity is communicated with the first air guide buffer cavity through a second air guide pipe, and the pressing device is connected with the upper piston rod through gear transmission.
Preferably, two gears which are meshed with each other are arranged between the upper piston rod and the lower piston rod and are respectively a driving gear and a driven gear which are correspondingly installed in a gear installation groove, the driving gear, the driven gear and the second reagent cavity are respectively located on two sides of the first reagent cavity, the driving gear and the driven gear are respectively rotatably connected inside the shell through shafts, the bottom of the upper piston rod is meshed with the driving gear, the top of the lower piston rod is meshed with the driven gear, and the driving gear is meshed with the pressing device.
Preferably, the pressing device includes the pressing device body, one side downwardly extending of the pressing device body form with the rack that drive gear engaged with, the inside slide rail that is equipped with of casing, be equipped with on the pressing device body and follow the slider that the slide rail reciprocated, the pressing device body is kept away from one side of rack is equipped with first draw-in groove, the casing is inside be equipped with first draw-in groove matched with protection lock, the last edge of pressing device body be the arc and with along flushing on the casing, the last edge width of pressing device body is greater than the width of pressing device body, and with casing thickness is unanimous.
Preferably, the protection lock is located inside the casing, the protection lock includes the catch bar, the last edge of catch bar with flush along on the casing, the catch bar lower extreme is equipped with the sliding block, one side of sliding block be equipped with first draw-in groove matched with bayonet lock, the opposite side of sliding block is equipped with bullet knot mechanism, bullet knot mechanism is the arc, bullet knot mechanism is kept away from the one end of sliding block is buckled downwards and to the direction of bayonet lock extends, bullet knot mechanism keeps away from the one end of sliding block is equipped with the latch, inside be equipped with of casing with latch matched with second draw-in groove and third draw-in groove.
Preferably, the lower piston rod comprises a lower piston core, a first plug head is tightly arranged on the periphery of the lower piston core, a first plug body is arranged on one side of the lower piston core, and a rack structure meshed with the driven gear is arranged at the top of the first plug body;
go up the piston rod and include the piston core, it sets up the second chock plug to go up the fastening of piston core periphery, it is equipped with the second cock body to go up piston core one side, the bottom of the second cock body be equipped with drive gear engaged with rack structure, the second cock body is equipped with the cavity structure, the casing is inside to be equipped with and runs through the spacing post of cavity structure.
Preferably, the reagent mixing part comprises a mixing tank and a liquid storage tank, a flow-limiting dam is arranged between the lower part of the mixing tank and the lower part of the liquid storage tank, the upper part of the mixing tank is communicated with the upper part of the liquid storage tank, the lower piston cavity is connected with the top of the mixing tank through a flow guide pipe, and the top of the liquid storage tank is communicated with the test strip mounting groove;
the sealing plug is passed through at test paper strip mounting groove top with the casing top is connected, the test paper strip mounting groove is located the avris setting of being close to the casing, the sealing plug is installed the corner position at casing top.
Preferably, the sealing plug includes the sealing plug body, the sealing plug body is pi shape (door arch shape), sealing plug body one side lower part is equipped with first buckle, the casing inside be equipped with first buckle matched with fourth draw-in groove, sealing plug body another side bottom is equipped with sealing member, sealing member follows the top of test paper strip mounting groove is inserted and is formed interference fit, the last edge of sealing plug body be the arc and with the casing is gone up along flushing, the width on the last edge of sealing plug body is greater than the width of sealing plug body, and with casing thickness is unanimous.
Preferably, the test strip mounting groove is made of a transparent material, and a transparent window is arranged at a position of the shell corresponding to the detection result reading position of the chromatographic test strip; the casing corresponds the position of first reagent cavity with the second reagent cavity is equipped with first application of sample hole and second application of sample hole respectively, first application of sample hole with the second application of sample hole is equipped with first application of sample stopper and second application of sample stopper of matched with.
The invention also provides a method for detecting nucleic acid by adopting the nucleic acid detection microfluidic device, which comprises the following steps:
(1) inserting a chromatographic test strip into the test strip mounting groove, extending the lower end of the chromatographic test strip into the bottom of the liquid storage tank, and mounting a sealing plug to seal the upper end of the test strip mounting groove;
(2) adding a first reagent into the first reagent cavity from the first sample adding hole, adding a second reagent into the second reagent cavity from the second sample adding hole, and respectively plugging the first sample adding hole and the second sample adding hole by using a first sample adding plug and a second sample adding plug, wherein the inner space of the shell is sealed and completely isolated from the outside; the first reagent is a buffer reagent, and the second reagent is a virus extract sample and an amplification reagent;
(3) heating the second reagent cavity for incubation; after the incubation is finished, the push rod is pushed, the pressing device is unlocked from the protection lock, the pressing device is pressed to move downwards, in the process that the pressing device moves downwards, the driving gear and the driven gear are driven to operate, the upper piston rod and the lower piston rod are synchronously driven to move towards the outer direction of the shell, so that the upper piston cavity and the lower piston cavity respectively form a vacuum cavity, along with the development of the movement, the vacuum cavity of the upper piston cavity and the vacuum cavity of the lower piston cavity are communicated with the first reagent cavity, the second reagent cavity, the first air guide tube, the second air guide tube, the first air guide buffer cavity and the second air guide buffer cavity simultaneously, due to the pressure compensation characteristic, the materials of the second reagent cavity and the first reagent cavity sequentially flow into the mixing tank through the lower piston cavity and the flow guide tube, the first reagent and the second reagent are fully mixed in the mixing tank, along with the rise of the liquid level in the mixing tank, in the mixed pond material overflowed the current-limiting dam and got into the liquid storage tank, the material in the chromatography test paper strip detected in the liquid storage tank, and CT line through the chromatography test paper strip is chromogenic, reads the testing result through the direct naked eye of transparent window.
Compared with the prior art, the invention has the following effects:
(1) the nucleic acid detection microfluidic device provided by the invention is highly integrated, integrates various detection reaction materials and detection units, is low in price, portable, easy to operate, quick in detection time, high in detection result accuracy and high in biological safety, and is used for nucleic acid detection.
(2) The inside of the kit (shell) is hermetically isolated from the external space, and due to the formation of the vacuum cavity, a negative pressure environment is formed inside relative to the outside. Therefore, biological components (such as nucleic acid amplification products) and the like in the kit are fully sealed in the inner cavity, the overflow risk of aerosol and the like is avoided, and the biological safety is ensured.
(3) The invention is suitable for nucleic acid detection of isothermal nucleic acid amplification technology based on the principles of RT-eRDA-LFD and the like, and is also suitable for a detection system which needs to reliably control multi-sample mixing, does not depend on a special instrument to realize detection and ensures biological safety.
(4) The second cavity can be pre-loaded with biological components required by nucleic acid cracking, extraction and amplification, and a user can directly add the sampling liquid into the second cavity after sampling the nasopharyngeal swab, so that the nucleic acid cracking and direct amplification are realized. The detection unit is a test strip, after amplification is completed, the device realizes mixing of various materials and triggering and control of a detection process in a self-driving mode, and all components are sealed inside the negative pressure device, so that the biological safety is fully guaranteed.
(5) The invention adopts the self-driving design based on the air pressure compensation balance principle, and can ensure that all materials in the cavity are fully driven to flow out, thereby ensuring the mixing proportion and sequence; the integrated test strip is used as a detection unit, so that the result output without the dependence of an external instrument can be realized; the method has low requirement on operation, is simple and easy to operate, and can be used for industrial production.
Drawings
FIG. 1 is a schematic diagram showing the internal structure of a microfluidic device for nucleic acid detection according to an embodiment of the present invention;
FIG. 2 is a schematic structural view of a front cover in an embodiment of the present invention;
FIG. 3 is a schematic structural view of a rear cover in an embodiment of the present invention;
FIG. 4 is a schematic structural diagram of a pressing device according to an embodiment of the present invention;
FIG. 5 is a schematic diagram of a dongle according to an embodiment of the present invention;
FIG. 6 is an exploded view of the lower piston rod in an embodiment of the present invention;
FIG. 7 is an exploded view of the upper piston rod in an embodiment of the present invention;
fig. 8 is a schematic view of the construction of a sealing plug in an embodiment of the invention;
FIG. 9 is a schematic diagram of a process for detecting nucleic acid using a nucleic acid detecting microfluidic device according to an embodiment of the present invention;
in the figure:
1-front cover, 2-back cover, 3-sealing plug, 3.1-sealing part, 3.2-first buckle, 3.3-sealing plug body, 4-pressing device, 4.1-pressing device body, 4.2-first clamping groove, 4.3-sliding block, 4.4-rack, 5-protection lock, 5.1-bayonet lock, 5.2-snap mechanism, 5.3-push rod, 5.4-sliding block, 5.5-latch, 6-upper piston rod, 6.1-second rack structure, 6.2-upper piston core, 6.3-second plug, 6.4-second annular groove structure, 6.5-second plug body, 6.6-cavity structure, 7-upper piston cavity, 8-driving gear, 9-lower piston cavity, 10-lower piston rod, 10.1-first rack structure, 10.2-lower piston core, 10.3-first plug head, 10.4-first annular groove structure, 10.5-first plug body, 11-driven gear, 12-draft tube, 13-flow-limiting dam, 14-liquid storage tank, 15-second air guide buffer cavity, 16-first reagent cavity, 17-second reagent cavity, 18-slide rail, 19-second air guide tube, 20-first air guide buffer cavity, 21-first air guide tube, 22-test paper strip mounting groove, 23-assembly pin, 24-mixing tank, 25-spacing column, 26-first sample adding hole, 27-second sample adding hole, 28-second clamping groove, 29-third clamping groove and 30-transparent window.
Detailed Description
Examples
The utility model provides a nucleic acid detection micro-fluidic device, which comprises a housin, the casing can be integrative preparation forms, also can be inside be the shell structure of two sides mutual symmetry complex, as shown in figure 2 and figure 3, be protecgulum 1 and back lid 2 respectively, protecgulum 1 is equipped with a plurality of groups equipment round pin 23 structures with the inside symmetry of back lid 2, with protecgulum 1 and back lid 2 equipment shaping in opposite directions for shell structure, the equipment round pin 23 equipment that will correspond is screwed up, through current technology (such as hasp or butt fusion etc.), form the inside cavity structure of casing, and mutual sealed faying face does not leak, as long as realize the inside structure of casing and sealed leak-free can.
As shown in fig. 1, a lower piston cavity 9 is arranged inside the casing, a lower piston rod 10 is connected with a piston in the lower piston cavity 9, the upper part of the lower piston cavity 9 is communicated with a first reagent cavity 16 and a second reagent cavity 17 respectively, the lower part of the lower piston cavity 9 is communicated with a reagent mixing component, one side of the reagent mixing component is communicated with a test strip mounting groove 22, a chromatography test strip is inserted into the test strip mounting groove 22 and extends into the reagent mixing component, an opening is arranged on one side of the casing and is provided with a pressing device 4, and the pressing device 4 extends into the casing and is connected with the lower piston rod 10 through gear transmission.
The top of the first reagent cavity 16 is communicated with the upper piston cavity 7, the piston in the upper piston cavity 7 is connected with the upper piston rod 6, the upper piston cavity 7 is communicated with the first air guide buffer cavity 20 through a first air guide pipe 21, the first air guide buffer cavity 20 is communicated with the second air guide buffer cavity 15, the second air guide buffer cavity 15 is communicated with one side of the lower piston cavity 9, the top of the second reagent cavity 17 is communicated with the first air guide buffer cavity 20 through a second air guide pipe 19, and the pressing device 4 is connected with the upper piston rod 6 through gear transmission.
Two gears which are meshed with each other are arranged between the upper piston rod 6 and the lower piston rod 10 and are respectively a driving gear 8 and a driven gear 11 which are correspondingly installed in the gear installation grooves, the driving gear 8, the driven gear 11 and the second reagent cavity 17 are respectively positioned at two sides of the first reagent cavity 16, the driving gear 8 and the driven gear 11 are respectively rotatably connected inside the shell through shafts, the bottom of the upper piston rod 6 is meshed with the driving gear 8, the top of the lower piston rod 10 is meshed with the driven gear 11, and the driving gear 8 is meshed with the pressing device 4.
As shown in fig. 4, the pressing device 4 includes a pressing device body 4.1, one side of the pressing device body 4.1 extends downwards to form a rack 4.4 meshed with the driving gear 8, a slide rail 18 is arranged inside the casing, a slider 4.3 capable of moving up and down along the slide rail 18 is arranged on the pressing device body 4.1, or a slider is arranged inside the casing, a slide rail capable of moving up and down along the slider is arranged on the pressing device body 4.1, only the up and down movement is realized, a first clamping groove 4.2 is arranged on one side of the pressing device body 4.1 far away from the rack 4.4, a protection lock 5 matched with the first clamping groove 4.2 is arranged inside the casing, the upper edge of the pressing device body 4.1 is arc-shaped and is flush with the upper edge of the casing, the upper edge width of the pressing device body 4.1 is greater than the width of the pressing device body 4.1, and is consistent with the thickness of the casing. One side of the pressing device body 4.1 extends downwards to form a rack 4.4 meshed with the driving gear 8 and used for being matched with the driving gear 8 and the driven gear 11, the driving gear 8 is driven to operate when the pressing device body 4 is pressed down, and the driving gear 8 drives the driven gear 11 to operate. The pressing device 4 is used for receiving a pressing action to drive the driving gear 8 to act, the sliding block 4.3 and the sliding rail 18 play a limiting role, so that the pressing device 4 moves downwards to drive the upper piston rod 6 to move (for example, move rightwards in the figure 1), the driving gear 8 drives the driven gear 11 at the same time, and the driven gear 11 drives the lower piston rod 10 to move (for example, move rightwards in the figure 1).
The protection lock 5 is located inside the casing, as shown in fig. 5, the protection lock 5 includes a push rod 5.3, the upper edge of the push rod 5.3 is flush with the upper edge of the casing, the lower end of the push rod 5.3 is provided with a sliding block 5.4, one side of the sliding block 5.4 is provided with a bayonet 5.1 matched with the first bayonet 4.2, the other side of the sliding block 5.4 is provided with a snap mechanism 5.2, the snap mechanism 5.2 is arc-shaped, one end of the snap mechanism 5.2 far away from the sliding block 5.4 is bent downwards and extends towards the direction of the bayonet 5.1, one end of the snap mechanism 5.2 far away from the sliding block 5.4 is provided with a latch 5.5, and the casing is internally provided with a second bayonet 28 and a third bayonet 29 matched with the latch 5.5.5.
The protection lock 5 is used to lock the pressing device 4 from being pressed before the detection of the activation. The elastic buckle mechanism 5.2 is arc-shaped, one end of the elastic buckle mechanism 5.2 is connected with the sliding block 5.4, the other end of the elastic buckle mechanism is bent downwards and extends towards the direction of the bayonet 5.1, a second clamping groove 28 and a third clamping groove 29 which are matched with the bayonet 5.5 are arranged inside the shell, the second clamping groove 28 is positioned on the left side of the third clamping groove 29, and the elastic buckle mechanism 5.2 is used for limiting the stroke range of the protection lock 5 so as to prevent misoperation. The push rod 5.3 is an operation surface, and the user uses the protection lock 5 by pushing the push rod 5.3. When the pushing rod 5.3 is pushed (for example, pushing to the right in fig. 1), the pushing rod 5.3 drives the sliding block 5.4 to slide along the inside of the housing, the latch 5.1 is pushed into the first slot 4.2 of the pressing device 4 to form locking, and at this time, the latch 5.5 is engaged with the third slot 29. When the pushing rod 5.3 is pushed (taking fig. 1 as an example, pushing to the left), the pushing rod 5.3 drives the sliding block 5.4 to slide along the inside of the housing, the latch 5.1 is pushed out of the first slot 4.2 of the pressing device 4, the locking is released, the latch 5.5 is pushed to be engaged with the second slot 28, and the latch 5.1 is released from the locking with the first slot 4.2 of the pressing device 4.
As shown in fig. 6, the lower piston rod 10 includes a lower piston core 10.2, a first plug 10.3 is fastened to an outer periphery of the lower piston core 10.2, a first plug 10.5 is disposed on one side of the lower piston core 10.2, a first rack structure 10.1 engaged with the driven gear 11 is disposed at a top of the first plug 10.5, an annular protrusion structure is disposed on an outer periphery of the lower piston core 10.2, a left lower corner of fig. 6 is a cross-sectional view of the first plug, a first annular groove structure 10.4 matched with the annular protrusion structure is disposed on an inner periphery of the first plug 10.3, the lower piston core 10.2 and the first plug 10.3 form a structure fastened and matched with each other, so as to prevent the first plug 10.3 from falling off during operation, and the first plug 10.3 is made of rubber. The limiting device of piston rod 10 can be set up down on the casing, for example, the lower part of piston rod 10 is established the spacing backing roll down, or is established the cavity structure down on the piston rod, is equipped with the spacing post that runs through this cavity structure in that the casing is inside, and spacing post supports and leads piston rod 10 down in piston rod 10 operation process down, as long as can realize down the spacing and the direction function of piston rod 10 can. The lower piston rod 10 is used for being assembled in the lower piston cavity 9, and has a sealing effect on the first reagent cavity 16, the second reagent cavity 17, the second air guide buffer cavity 15 and the draft tube 12, so as to block and isolate the cavities and substances in the cavities.
As shown in fig. 7, the upper piston rod 6 includes an upper piston core 6.2, a second plug 6.3 is fastened to the periphery of the upper piston core 6.2, a second plug 6.5 is arranged on one side of the upper piston core 6.2, a second rack structure 6.1 meshed with the driving gear 8 is arranged at the bottom of the second plug 6.5, a cavity structure 6.6 is arranged on the second plug 6.5, and a limit column 25 penetrating through the cavity structure 6.6 is arranged inside the housing. Go up piston core 6.2 periphery and set up cyclic annular protruding structure, the figure in the lower left corner of figure 7 is second chock plug 6.3's cross-sectional view, second chock plug 6.3 internal week be equipped with cyclic annular protruding structure matched with second annular groove structure 6.4, go up piston core 6.2 and second chock plug 6.3 formation fastening complex structure, prevent that second chock plug 6.3 from droing at the operation in-process, the material of second chock plug 6.3 is the rubber material, second chock plug 6.5 is equipped with cavity structure 6.6, the inside spacing post 25 that runs through cavity structure 6.6 that is equipped with of casing, spacing post 25 supports and leads last piston rod 6 at last piston rod 6 operation in-process. The upper piston rod 6 is used for being assembled in the upper piston cavity 7, and sealing the first reagent cavity 16 and the first air duct 21 to block and isolate the first reagent cavity 16 and substances in the first reagent cavity 16.
The reagent mixing part comprises a mixing tank 24 and a liquid storage tank 14, a flow limiting dam 13 is arranged between the lower part of the mixing tank 24 and the lower part of the liquid storage tank 14, the upper part of the mixing tank 24 is communicated with the upper part of the liquid storage tank 14, a lower piston cavity 9 is connected with the top of the mixing tank 24 through a flow guide pipe 12, and the top of the liquid storage tank 14 is communicated with a test strip mounting groove 22.
The top of the test strip mounting groove 22 is connected with the top of the shell through the sealing plug 3, the test strip mounting groove 22 is arranged on the side close to the shell, and the sealing plug 3 is mounted at the corner of the top of the shell.
As shown in fig. 8, the sealing plug 3 includes a sealing plug body 3.3, the sealing plug body 3.3 is pi shape, one side lower part of the sealing plug body 3.3 is equipped with a first buckle 3.2, the inside fourth draw-in groove that matches with first buckle 3.2 that can be equipped with of casing, sealing plug body 3.3 another side bottom is equipped with sealing member 3.1, sealing member 3.1 inserts from the top of test paper strip mounting groove 22 and forms interference fit, the last edge of sealing plug body 3.3 is the arc and flushes with the casing upper edge, the width on the last edge of sealing plug body 3.3 is greater than the width of sealing plug body 3.3, and is unanimous with casing thickness. After chromatography test paper strip is packed into test paper strip mounting groove 22, install sealing plug 3 in the corner position at the casing top from top to bottom, sealing member 3.1 inserts from the top of test paper strip mounting groove 22 this moment and forms interference fit and form sealedly to test paper strip mounting groove 22, in order to reach better sealed effect, can be equipped with in the casing inside with first buckle 3.2 matched with fourth draw-in groove, first buckle 3.2 is fixed with fourth draw-in groove block, prevents that sealing plug 3 from droing.
The test paper strip mounting groove 22 and the casing all adopt transparent material, perhaps the casing corresponds the testing result reading position department of chromatography test paper strip and is equipped with transparent window 30, and this transparent window 30 is used for providing complete clear observation face interface and distinguishes for the user and read the testing result, and this transparent window 30 design has the round outer edge for combine each other and realize sealedly (modes such as accessible butt fusion or sealing washer) with the casing. The casing is provided with a first sample adding hole 26 and a second sample adding hole 27 at positions corresponding to the first reagent cavity 16 and the second reagent cavity 17, respectively, and the first sample adding hole 26 and the second sample adding hole 27 are provided with a first sample adding plug and a second sample adding plug which are matched with each other. The first and second plug co-act to achieve a reliable seal of the first and second reagent chambers 16, 17. The first reagent chamber 16 may be pre-filled with the desired reagents during manufacture and during transport turnaround prior to subsequent delivery to the end user to ensure stability and preservation of the filled substance.
The method for detecting the nucleic acid by adopting the nucleic acid detection microfluidic device comprises the following steps:
(1) inserting a chromatographic test strip into the test strip mounting groove, extending the lower end of the chromatographic test strip into the bottom of the liquid storage tank, and mounting a sealing plug to seal the upper end of the test strip mounting groove;
(2) adding a first reagent into the first reagent cavity from the first sample adding hole, adding a second reagent into the second reagent cavity from the second sample adding hole, and respectively plugging the first sample adding hole and the second sample adding hole by using a first sample adding plug and a second sample adding plug, wherein the inner space of the shell is sealed and completely isolated from the outside; the first reagent is a buffer reagent, and the second reagent is a virus extract sample and an amplification reagent; the chromatographic test strip adopted by the invention is a Lateral flow chromatographic test strip (LFD); the amplification reagent and the buffer reagent are conventional and commercially available in the field;
(3) heating the second reagent cavity for incubation; the incubation device can be any device capable of heating a specific part, for example, an incubation device with a plurality of carrying mechanisms can be used for mounting a plurality of reagent kits, and a second cavity (e.g., a wire frame part at the lower left corner of the rightmost drawing in the first row of fig. 9) is subjected to heating incubation under the following conditions: heating at the constant temperature of 37-42 ℃ for 20-40 minutes, and preferably prompting a user through an LED or buzzer by the incubation device after the heating incubation process is finished.
After the incubation is finished, the push rod is pushed, the pressing device is unlocked from the protection lock, the pressing device is pressed to move downwards, in the process that the pressing device moves downwards, the driving gear and the driven gear are driven to operate, the upper piston rod and the lower piston rod are synchronously driven to move towards the outer direction of the shell, so that the upper piston cavity and the lower piston cavity respectively form a vacuum cavity, along with the development of the movement, the vacuum cavity of the upper piston cavity and the vacuum cavity of the lower piston cavity are communicated with the first reagent cavity, the second reagent cavity, the first air guide tube, the second air guide tube, the first air guide buffer cavity and the second air guide buffer cavity simultaneously, due to the pressure compensation characteristic, the materials of the second reagent cavity and the first reagent cavity sequentially flow into the mixing tank through the lower piston cavity and the flow guide tube, the first reagent and the second reagent are fully mixed in the mixing tank, along with the rise of the liquid level in the mixing tank, in the mixed pond material overflowed the current-limiting dam and got into the stock solution pond, the chromatography test paper strip detected the material in the stock solution pond, and CT line through the chromatography test paper strip shows colour, and direct naked eye reads testing result.
Fig. 9 is a schematic diagram of the process of nucleic acid detection by the micro-fluidic nucleic acid detection device of the invention, in which the upper left row of the diagram is to add a sample, the middle diagram of the upper row is to cover a sealing plug, the right row of the diagram of the upper row is to heat a second reagent chamber inside a dotted line frame, the right row of the diagram of the lower right row is to pull out a safety lock, the middle diagram of the lower row is to press down the pressing device, and the left row of the diagram of the lower left row is to read the detection result from one side (a transparent observation window is arranged at the position corresponding to the detection result reading position of the chromatographic test strip on one side of the housing).
Taking a nucleic acid detection scene based on an RT-eRDA-LFD isothermal nucleic acid amplification technology as an example, the application process of the invention is as follows:
(1) assembling and preparing a kit (shell):
(a) the casing adopts the structure that two sides are mutually supported, is upper cover and lower cover respectively, with upper cover and lower cover through prior art (like hasp or butt fusion etc.), the kit main part is assembled shaping in opposite directions to form the inside cavity structure of kit, and the sealed nothing of faying face each other leaks.
(b) Inserting a chromatographic test strip into the test strip mounting groove, extending the lower end of the chromatographic test strip into the bottom of the liquid storage tank, and mounting a sealing plug to seal the upper end of the test strip mounting groove;
(c) adjusting the tooth positions of each component by a pressing device, a driving gear, a driven gear, a lower piston rod, a lower piston cavity, an upper piston rod and an upper piston cavity, so that the tooth positions are mutually meshed and matched, the upper edge of the pressing device is flush with the upper edge of a kit, a first clamping groove on the pressing device is interlocked with a clamping pin on a protection lock, and a clamping tooth on the protection lock is clamped with a third clamping groove in a shell;
(2) the testing process comprises the following steps:
(a) adding a first reagent into the first reagent cavity from the first sample adding hole, adding a second reagent into the second reagent cavity from the second sample adding hole, and respectively plugging the first sample adding hole and the second sample adding hole by using a first sample adding plug and a second sample adding plug, wherein the inner space of the shell is sealed and completely isolated from the outside; the first reagent is a buffer reagent, and the second reagent is a virus extract sample and an amplification reagent; so far, the inner space of the reagent box cavity is sealed and completely isolated from the outside.
(b) The kit is then placed on the incubation device, the incubation device is activated and incubation is performed at the second reagent chamber.
(c) After the incubation is finished, the push rod is pushed (leftwards as shown in fig. 1), the push rod drives the sliding block to slide along the inside of the shell, the latch on the protection lock is not clamped with the third clamping groove any more, the latch on the protection lock is pushed to be clamped with the second clamping groove, the clamping pin is pushed out of the first clamping groove of the pressing device, the locking is released, and the pressing device is pressed downwards.
(d) In the descending process of the pressing device, the driving gear and the driven gear are driven to run to synchronously drive the upper piston rod and the lower piston rod to move towards the outer direction of the shell (rightward movement as shown in figure 1), so that the upper piston cavity and the lower piston cavity respectively form a vacuum cavity, and along with the development of the movement, the vacuum cavity of the upper piston cavity and the vacuum cavity of the lower piston cavity simultaneously form a first reagent cavity, a second reagent cavity and a first air duct, the second air duct, the first air guide buffer cavity and the second air guide buffer cavity are communicated, at the moment, due to the pressure compensation characteristic, materials in the second reagent cavity and the first reagent cavity flow into the lower piston cavity in sequence within the time of pressure compensation in the cavity until the pressure is balanced and then flow into the mixing pool through the flow guide pipe, and the first reagent and the second reagent are fully mixed in the mixing pool;
(e) it should be noted that, in the present invention, especially considering the opening flow sequence of the second reagent chamber and the first reagent chamber during the movement of the upper piston rod and the lower piston rod, the second reagent chamber is designed to be opened first, and the first reagent chamber is designed to be opened later, so as to ensure that the substance sample (such as the buffer reagent) in the first reagent chamber and the substance sample (such as the nucleic acid amplification product) in the second reagent chamber are fully diluted and fused. Similar concepts can be applied to other usage scenarios by adjusting the size and arrangement position of the related structures, which are not described in detail herein.
(f) After the mixture flows into the mixing pool, the flow limiting dam prevents the mixed liquid from contacting the chromatography test paper in time, and the mixed liquid overflows the flow limiting dam along with the rise of the liquid level in the mixing pool and contacts the chromatography test paper and triggers the detection before entering the liquid storage pool. The purpose of this design lies in guaranteeing that the mixture has further homogeneous mixing of abundant time after flowing into the mixing tank, then detects, and detection effect is more accurate.
(g) And (3) developing the color of the CT line of the chromatographic test strip, and directly reading the detection result by naked eyes of a user.
(h) After the detection is finished, the user takes the kit down from the incubation device, and the kit is put into the original packaging sealing bag of the kit for recycling.
Because the inside of the reagent box is sealed and isolated from the external space, and because of the formation of the vacuum cavity, the inside forms a negative pressure environment relative to the outside. Therefore, biological components (such as nucleic acid amplification products) and the like in the kit are fully sealed in the internal cavity, the risk of overflowing of aerosol and the like is avoided, and the biological safety is ensured.
The design idea and the technical points disclosed in the present invention are not limited to nucleic acid detection. Any scenario of multi-sample mixed detection has the opportunity to benefit from the information disclosed in the present invention.
Claims (10)
1. A nucleic acid detection microfluidic device, characterized in that: the test paper box comprises a housin, the inside piston cavity that is equipped with of casing, piston rod under the piston cavity internal piston connection down, the upper portion of piston cavity communicates first reagent cavity and second reagent cavity respectively down, the lower part intercommunication reagent mixing component of piston cavity down, one side and the test paper strip mounting groove intercommunication of reagent mixing component, insert the chromatography test paper strip in the test paper strip mounting groove, the chromatography test paper strip extends into inside the reagent mixing component, casing one side is equipped with the opening, the opening is equipped with press device, press device extends into inside and through gear drive connection of casing piston rod down.
2. The nucleic acid detecting microfluidic device according to claim 1, wherein: the top of the first reagent cavity is communicated with an upper piston cavity, the piston in the upper piston cavity is connected with an upper piston rod, the upper piston cavity is communicated with a first air guide buffer cavity through a first air guide pipe, the first air guide buffer cavity is communicated with a second air guide buffer cavity, the second air guide buffer cavity is communicated with one side of the lower piston cavity, the top of the second reagent cavity is communicated with the first air guide buffer cavity through a second air guide pipe, and the pressing device is connected with the upper piston rod through gear transmission.
3. The nucleic acid detecting microfluidic device according to claim 2, wherein: two gears which are meshed with each other are arranged between the upper piston rod and the lower piston rod and are respectively a driving gear and a driven gear which are correspondingly installed in the gear installation grooves, the driving gear, the driven gear and the second reagent cavity are respectively located on two sides of the first reagent cavity, the driving gear and the driven gear are respectively connected inside the shell in a rotating mode through shafts, the bottom of the upper piston rod is meshed with the driving gear, the top of the lower piston rod is meshed with the driven gear, and the driving gear is meshed with the pressing device.
4. The nucleic acid detecting microfluidic device according to claim 3, wherein: the pressing device comprises a pressing device body, one side of the pressing device body extends downwards to form a rack meshed with the driving gear, a sliding rail is arranged inside the shell, a sliding block which can move up and down along the sliding rail is arranged on the pressing device body, a first clamping groove is formed in one side of the rack, a protective lock matched with the first clamping groove is arranged inside the shell, the upper edge of the pressing device body is arc-shaped and is flushed with the upper edge of the shell, and the upper edge of the pressing device body is wider than the width of the pressing device body and is consistent with the thickness of the shell.
5. The nucleic acid detecting microfluidic device according to claim 4, wherein: the fender lock is located inside the casing, the fender lock includes the catch bar, the last edge of catch bar with along flushing on the casing, the catch bar lower extreme is equipped with the sliding block, one side of sliding block be equipped with first draw-in groove matched with bayonet lock, the opposite side of sliding block is equipped with bullet knot mechanism, bullet knot mechanism is the arc, bullet knot mechanism is kept away from the one end of sliding block is buckled downwards and to the direction of bayonet lock extends, bullet knot mechanism keeps away from the one end of sliding block is equipped with the latch, inside being equipped with of casing with latch matched with second draw-in groove and third draw-in groove.
6. The nucleic acid detecting microfluidic device according to claim 3, wherein: the lower piston rod comprises a lower piston core, a first plug head is fixedly arranged on the periphery of the lower piston core, a first plug body is arranged on one side of the lower piston core, and a rack structure meshed with the driven gear is arranged at the top of the first plug body;
go up the piston rod and include the piston core, it sets up the second chock plug to go up the fastening of piston core periphery, it is equipped with the second cock body to go up piston core one side, the bottom of second cock body be equipped with drive gear engaged with's rack structure, the second cock body is equipped with the cavity structure, the casing is inside to be equipped with to run through the spacing post of cavity structure.
7. The nucleic acid detecting microfluidic device according to claim 1, wherein: the reagent mixing component comprises a mixing tank and a liquid storage tank, a flow-limiting dam is arranged between the lower part of the mixing tank and the lower part of the liquid storage tank, the upper part of the mixing tank is communicated with the upper part of the liquid storage tank, the lower piston cavity is connected with the top of the mixing tank through a flow guide pipe, and the top of the liquid storage tank is communicated with the test strip mounting groove;
the test paper strip mounting groove top pass through the sealing plug with the casing top is connected, the test paper strip mounting groove is located the avris setting of being close to the casing, the sealing plug is installed the corner position at casing top.
8. The nucleic acid detecting microfluidic device according to claim 7, wherein: the sealing plug comprises a sealing plug body, the sealing plug body is n-shaped, a first buckle is arranged on the lower portion of one side of the sealing plug body, a fourth clamping groove matched with the first buckle is formed in the shell, a sealing part is arranged at the bottom of the other side of the sealing plug body, the sealing part is inserted into the top of the test strip mounting groove to form interference fit, the upper edge of the sealing plug body is arc-shaped and is flush with the upper edge of the shell, and the width of the upper edge of the sealing plug body is greater than that of the sealing plug body and is consistent with the thickness of the shell.
9. The nucleic acid detecting microfluidic device according to claim 1, wherein: the test strip mounting groove is made of transparent materials, and a transparent window is arranged at the position, corresponding to the detection result reading position of the chromatographic test strip, of the shell; the casing corresponds the position of first reagent cavity with the second reagent cavity is equipped with first application of sample hole and second application of sample hole respectively, first application of sample hole with the second application of sample hole is equipped with first application of sample stopper and second application of sample stopper of matched with.
10. A method for detecting nucleic acid using the nucleic acid detecting microfluidic device according to any one of claims 1 to 9, wherein: the method comprises the following steps:
(1) inserting the chromatographic test strip into the test strip mounting groove, extending the lower end of the chromatographic test strip into the bottom of the liquid storage tank, and mounting a sealing plug to seal the upper end of the test strip mounting groove;
(2) adding a first reagent into the first reagent cavity from the first sample adding hole, adding a second reagent into the second reagent cavity from the second sample adding hole, and respectively plugging the first sample adding hole and the second sample adding hole by using a first sample adding plug and a second sample adding plug, wherein the inner space of the shell is sealed and completely isolated from the outside; the first reagent is a buffer reagent, and the second reagent is a virus extract sample and an amplification reagent;
(3) heating the second reagent cavity for incubation; after the incubation is finished, the push rod is pushed, the pressing device is unlocked from the protection lock, the pressing device is pressed to move downwards, in the process that the pressing device moves downwards, the driving gear and the driven gear are driven to operate, the upper piston rod and the lower piston rod are synchronously driven to move towards the outer direction of the shell, so that the upper piston cavity and the lower piston cavity respectively form a vacuum cavity, along with the development of the movement, the vacuum cavity of the upper piston cavity and the vacuum cavity of the lower piston cavity are communicated with the first reagent cavity, the second reagent cavity, the first air guide tube, the second air guide tube, the first air guide buffer cavity and the second air guide buffer cavity simultaneously, due to the pressure compensation characteristic, the materials of the second reagent cavity and the first reagent cavity sequentially flow into the mixing tank through the lower piston cavity and the flow guide tube, the first reagent and the second reagent are fully mixed in the mixing tank, along with the rise of the liquid level in the mixing tank, in the mixed pond material overflowed the current-limiting dam and got into the stock solution pond, the chromatography test paper strip detected the material in the stock solution pond, and CT line through the chromatography test paper strip shows colour, and direct naked eye reads testing result.
Priority Applications (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210365691.XA CN114931986B (en) | 2022-04-08 | 2022-04-08 | Nucleic acid detection micro-fluidic device and nucleic acid detection method thereof |
| PCT/CN2022/113382 WO2023193386A1 (en) | 2022-04-08 | 2022-08-18 | Nucleic acid detection microfluidic device and nucleic acid detection method therefor |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210365691.XA CN114931986B (en) | 2022-04-08 | 2022-04-08 | Nucleic acid detection micro-fluidic device and nucleic acid detection method thereof |
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| CN114931986A true CN114931986A (en) | 2022-08-23 |
| CN114931986B CN114931986B (en) | 2022-12-27 |
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| CN202210365691.XA Active CN114931986B (en) | 2022-04-08 | 2022-04-08 | Nucleic acid detection micro-fluidic device and nucleic acid detection method thereof |
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| WO (1) | WO2023193386A1 (en) |
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| WO2023193386A1 (en) | 2023-10-12 |
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