CN206127278U - Nucleic acid extraction , increase and detection integrated device - Google Patents
Nucleic acid extraction , increase and detection integrated device Download PDFInfo
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- CN206127278U CN206127278U CN201621127212.7U CN201621127212U CN206127278U CN 206127278 U CN206127278 U CN 206127278U CN 201621127212 U CN201621127212 U CN 201621127212U CN 206127278 U CN206127278 U CN 206127278U
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- nucleic acid
- paper substrate
- acid extraction
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Abstract
The utility model discloses a nucleic acid extraction, increase and detection integrated device, photocuring resin upper cover and base including the printing of 3D printing technique, the upper cover includes injection hole, control button, U type clamp fixed orifices and test paper observation window as a result, base first part moulding piece includes paper base nucleic acid extraction module, isothermal increase module and side flow test paper detection module. Wherein paper base nucleic acid extraction module includes reservoir, lysing channel and cleaning duct, nuclear acid adsorption paper and the pad that absorbs water, isothermal increase module includes heating plate, U type clamp, button cell and PMMA plastic slab. Test paper detection module includes test paper strip and chromatography liquid hold up tank. It is last, withhold device equipment completion on the base with the upper cover. Paper base nucleic acid extraction, increase and detection integrated device need not external device and professional staff, have low cost, easy operation, fast, easily the advantage such as carry, be applicable to the molecular diagnosis field, especially in remote districts that the resource is deficient.
Description
【Technical field】
This utility model is related to a kind of nucleic acid extraction, amplification and detection integrating device.
【Background technology】
Nucleic acid has been widely used in medical diagnosis on disease, food safety detection and environmental monitoring neck as a kind of molecular marker
Domain.Extraction, three parts of amplification and detection of nucleic acid are generally comprised based on the diagnosis of nucleic acid.Traditional method is mainly in laboratory
Large-scale instrument and technical professional are depended on, there is cost intensive, the shortcomings of taking time and effort.And in infectious disease frequency
Send out, food-safety problem is projected, the serious remote districts of environmental pollution, these traditional methods are not simultaneously applied to, therefore seriously indulge in
Wu Liao this areas people are to the Clinics and Practices of disease, the detection of food and the monitoring of environment and improvement.
In recent years, with the development of paper substrate micro-fluidic technologies, low cost, miniaturization, paper substrate easy to carry, simple to operate
Immediately detection means is day by day paid close attention to by people.At present, the extracting method of paper substrate has been (such as on the market：Whatman companies are sent out
The method of nucleic acid is extracted on bright paper (such as：FTA cards etc.)) and paper substrate nucleic acid amplification and test paper detecting method (such as：BioHelix
The the BESt cassette products of invention, including amplification groove and sensing chamber).They are required for the extraction and expansion for carrying out complexity
Increase operation, further rely on the large-scale instrument of laboratory (such as：Thermal cycler, refrigerator), however it remains not readily portable, operation is multiple
The defects such as miscellaneous, cost intensive.Quick, simple, portable detection of nucleic acids can not be realized.Still it is not suitable for scarcity of resources area to enter
Row molecular diagnosis.
【Utility model content】
The purpose of this utility model is the defect existed for existing paper substrate nucleic acid extraction, amplification and detection method, is carried
For a kind of nucleic acid extraction, amplification and detection integrating device, the installation cost is cheap, simple to operate, easy to carry, and detection time
It is short, operated without the need for professional and technical personnel, efficiency high is applied widely.
To reach above-mentioned purpose, this utility model is employed the following technical solutions and is achieved：
Nucleic acid extraction, amplification and detection integrating device, including the upper lid and pedestal that fasten up and down, are provided with paper substrate on pedestal
Nucleic acid extraction module, paper substrate nucleic acid isothermal amplification module and detection paper module；Entering for Deca sample is offered on upper lid
Sample hole, control knob and test paper result observation window；The U-shaped folder of movement is provided with paper substrate nucleic acid isothermal amplification module,
The side of upper lid is offered for fixing the fixing hole of U-shaped folder；U-shaped folder includes that what upper and lower two-layer was closely superimposed can lead
, to drag end, the other end is scraps of paper area for the sheet metal of heat, wherein one end；The upper strata in scraps of paper area is inlaid with the dependence unwindase of lyophilizing
Isothermal duplication reagent the scraps of paper, be inlaid with nucleic acid absorption paper on the relevant position of lower floor, drag end and stretched out by fixing hole；
During nucleic acid extraction, the end of the paper substrate cracking passage in paper substrate nucleic acid extraction module is contacted with nucleic acid absorption paper,
And the scraps of paper are not contacted with nucleic acid absorption paper；
During isothermal duplication, the heating plate in paper substrate nucleic acid isothermal amplification module is contacted with nucleic acid absorption paper and the scraps of paper, core
The PMMA plates of Aerosol Pollution when avoiding nucleic acid amplification of sealing are provided with sour adsorption paper and the scraps of paper；
During detection, nucleic acid absorption paper is located between the test strips in detection paper module and chromatographic solution holding tank, and with two
Person contacts.
This utility model is further improved by：
The paper substrate nucleic acid extraction module includes cleanout fluid liquid storage tank and lysate liquid storage tank, the lower end of cleanout fluid liquid storage tank
It is connected with the entrance of paper substrate flushing channel, the entrance that the lower end of lysate liquid storage tank cracks passage with paper substrate is connected；Paper substrate is cracked
The lower section of passage arranges adsorptive pads；The end of paper substrate flushing channel arranges paper substrate valve, and in cleaning, paper substrate valve is split with paper substrate
Solution passage contacts, and paper substrate flushing channel is connected with paper substrate cracking passage.
Caulking gum and sponge are provided with the cleanout fluid liquid storage tank and lysate liquid storage tank, sponge is located at sealing rubber
The lower section of glue.
The paper substrate nucleic acid isothermal amplification module also includes the power supply being connected with heating plate, and heating plate adopts ceramic heat
Piece.
The detection paper module includes Sidestream chromatography test strips and chromatographic solution holding tank, and chromatographic solution holding tank is located at effluent
Below the end of chromatography paper slip；Sidestream chromatography paper slip is corresponding with the position of the test paper result observation window on upper lid.
The Sidestream chromatography paper slip includes gum plate, sample pad, pad and the cellulose acetate being cascading
Film；It is placed with chromatographic solution holding tank for storing the sponge of chromatographic solution.
Compared with prior art, this utility model has the advantages that：
This utility model has the advantages that without the need for additional instruments, with low cost, easy to carry, simple to operate, quick.1) profit
Computer software design apparatus structure is used, nucleic acid extraction, amplification and detection is integrated in a device, sample is realized and is entered
As a result the integration detection for going out；2) this device only needs simple four operations：It is U-shaped that sample-adding → control knob → dragging is extracted in pressing
Clip to that amplification indicatrix → dragging is U-shaped to clip to reagent paper indicatrix, whole detection only needs to 77 minutes (detect needs in traditional experiment room
4-5 hours), operated without the need for professional and technical personnel, solved problem simple to operate, quick.3) paper substrate extraction module and
Detection paper module is combined the storage problem for solving extracting solution and chromatographic solution with sponge.Meanwhile, using freeze drying technology by tHDA
Amplifing reagent is stored on paper, without the need for cryogenic refrigerator.In addition, amplification heating provides electricity by button cell using ceramic heating flake
Source, without the need for large-scale heater.And in paper substrate module, liquid is flowed by capillary force forward, without the need for external pump, therefore with
Upper method solves the problems, such as easy to carry.4) part and chemical reaction reagent selected by this device is with low cost, solves
The problem of reduces cost.
【Description of the drawings】
Fig. 1 is paper substrate nucleic acid extraction described in the utility model, amplification and detects integrating device superstructure schematic diagram；
Fig. 2 is paper substrate nucleic acid extraction described in the utility model, amplification and detects integrating device base construction schematic diagram；
Fig. 3 is paper substrate nucleic acid extraction described in the utility model, amplification and detects that (paper substrate extracts mould to integrating device integration module
Block, paper substrate nucleic acid amplification module and detection paper module) structural representation；
Fig. 4 is paper substrate nucleic acid extraction described in the utility model, amplification and detects integrating device pedestal and its integration module
Schematic diagram；
Fig. 5 is the testing result that variable concentrations Salmonella is detected using this utility model.
Wherein, 1- sample holes；2- control knobs；3- fixing holes；4- test paper result observation windows；5- cleanout fluid liquid storage tanks；6-
Lysate liquid storage tank；7- liquid storage holes；8- paper substrates crack passage；9- paper substrate flushing channels；10- adsorptive pads；11- nucleic acid absorption paper；
The 12- scraps of paper；13- heating plates；14- power supplys；15-PMMA plates；16-U types are pressed from both sides；17- chromatographic solution holding tanks；18- test strips.
【Specific embodiment】
Below in conjunction with the accompanying drawings this utility model is described in further detail：
Referring to Fig. 1-4, this utility model paper substrate nucleic acid extraction, amplification and detection integrating device, including using 3D printing skill
The integration module on the upper lid of light-cured resin material, pedestal and pedestal that art is printed, upper lid is withheld on pedestal.Upper lid bag
Include sample holes 1, the control knob 2 being fixed on lid, the hole 3 of fixed " U " type folder and test paper result observation window 4.Collect on pedestal
Include paper substrate nucleic acid extraction module, paper substrate nucleic acid isothermal amplification module and test paper detection module into module.
Paper substrate nucleic acid extraction module includes the lysate liquid storage of the cleanout fluid liquid storage tank 5 and a diameter of 9mm of a diameter of 12mm
Pond 6, cleanout fluid liquid storage tank 5 and lysate liquid storage tank 6 are constituted by sponge and caulking gum, and, under, caulking gum is upper for sponge；
The position that wherein each liquid storage tank is connected with paper passage be provided with size be 5mm × 2mm liquid storage hole 7, liquid storage hole 7 respectively with paper substrate
Cracking passage and paper substrate flushing channel are connected；Cracking passage 8 one end is connected with lysate liquid storage tank 6, one end and nucleic acid absorption paper
It is connected, 9 one end of flushing channel is connected with cleanout fluid liquid storage tank 5, and the other end is made up of paper substrate valve, is connected with paper substrate cracking passage
Realize many more manipulations；Nucleic acid absorption paper 11 is fixed on the lower floor of U-shaped folder and is connected with one end of paper substrate cracking passage, and U-shaped folder
Upper strata be interposed between above by a girder, during nucleic acid extraction not with nucleic acid absorption paper joint；Adsorptive pads 10 exist
Below nucleic acid absorption paper.
Paper substrate cracks the common filter paper of material selection of passage 8 and paper substrate flushing channel 9, and its sample introduction zone is using a diameter of
The filter paper of 5mm～8mm, the size of paper substrate cracking passage 8 is 25mm × 5mm, and the size of paper substrate flushing channel 9 is 10mm × 5mm.
Nucleic acid absorption paper selects glass fibre, and its size is 5mm × 7mm, and adsorptive pads select absorbent paper, and its size is 30mm × 20mm.
Paper substrate nucleic acid isothermal amplification module includes ultra-thin ceramic heating flake, the removable U-shaped folder 16 for being inlaid with paper, provides
The battery of power supply 14 and the plastic plate being covered on heating plate.Wherein U-shaped folder material be two layers can the sheet metal of heat conduction closely fold
It is added together, its size is 8mm × 100mm.The osculum of 5mm × 7mm is provided with U-shaped folder 10mm wherein, upper strata is inlaid with
12 size of the scraps of paper of the isothermal duplication tHDA reagents of the dependence unwindase of lyophilizing is 5mm × 7mm, and lower floor relevant position is inlaid with core
11 size of sour adsorption paper is 5mm × 7mm, meanwhile, there is the indicatrix of two black on the sheet metal of upper strata, respectively distance drags end
The amplification indicatrix and distance of 47mm drag the detection indicatrix of end 62mm；The 3V of a diameter of 12mm of Selection of Battery of power supply is provided
Button cell, when being expanded, lays battery trench bottom on pedestal and is provided with the osculum of one 5mm × 2mm for placing absolutely
Edge paper is covered in the plastic plate preferred dimension on heating plate for 25mm × 30mm's blocking the conducting between power supply and electric wire
PMMA plates 15, this plate are in order to avoid Aerosol Pollution during nucleic acid amplification.
Detection paper module includes effluent journey analysis test strips 18 and chromatographic solution accumulator tank 17.Its effluent journey analysis reagent paper is by there is chi
The acetic acid of very little gum plate for 60mm × 3mm, the sample pad of 15mm × 3mm, the pad of 10mm × 3mm and 20mm × 3mm is fine
The plain film layer of dimension is folded to be constituted, and absorbent paper is size for the T-shape of 6mm × 14mm, chromatographic solution holding tank placed size for 10mm ×
The sponge of 5mm × 5mm is used for storing chromatographic solution.
This utility model is as follows preparing：
(1) using the structure of lid and pedestal in computer software design, the upper lid for designing and base construction are imported into 3D
In print software, printed with 3D printer respectively.
(2) sponge, rubber gasket, PMMA plates and the insulating hydrophobic paper of corresponding size are prepared with laser cutting machine；
(3) liquid storage sponge is soaked in corresponding liquid respectively makes sponge hydraulically full；
(4) size according to paper substrate module, prepares the paper substrate passage of corresponding size with laser cutting machine, is prepared with card punch
Sample introduction zone filter paper and nucleic acid absorption area glass fiber sheets.
(5) copper sheet of U-shaped folder is prepared with wire cutting machine tool.
(6) the isothermal duplication tHDA reagent scraps of paper for relying on unwindase are prepared with cryodesiccated method.
(7) use nano gold mark detection probe, assembling to paste reagent paper Related Component, effluent journey analysis reagent paper is prepared with cutting machine
Bar.
(8) by the corresponding component liquid storage sponge in paper substrate nucleic acid extraction module, caulking gum, paper substrate passage, paper substrate nucleic acid
The corresponding component ceramic heating flake of amplification module, button cell, insulating hydrophobic paper, PMMA plates are inlaid with nucleic acid absorption paper and jelly
The corresponding component effluent journey analysis of the U-shaped folder and detection paper module of the scraps of paper of the isothermal duplication tHDA reagents of dry dependence unwindase
Test strips and chromatographic solution storage sponge are respectively placed in corresponding position.
(9) each part in integrating device is pasted on the relevant position of pedestal respectively；
(10) upper lid and pedestal are assembled to form paper substrate nucleic acid extraction, amplification and detect integrating device.
This utility model using method is as follows：
(1) sample drop is added at sample holes 1.
(2) control knob 2 of lid is pressed, after waiting 2 minutes, extraction is completed.
(3) drag U-shaped clipping to handss to expand at instruction line position, now the scraps of paper on U-shaped folder are in close contact with heating plate,
And then the insulating paper at battery case is taken out, starts nucleic acid amplification, after waiting 60 minutes, amplification is completed.
(4) U-shaped clipping to is dragged again detects that now the scraps of paper area on U-shaped folder makes the chromatographic solution of separation at instruction line position
Sponge is connected with the sample pad of test strips, and sample is chromatographed in test strips by capillary force, after waiting 15 minutes, has been detected
Into.
(5) testing result is observed by test paper result observation window, when control zone and detection zone are redness, is judged to
Positive findingses, when control zone is red, result of determination is feminine gender.
The extraction flow process of this utility model sample：Sample is entered from sample holes Deca, control knob is pressed, is so controlled
Button can and sealing ring extrude sponge together, the liquid in sponge is flowed out from the aperture being connected with paper passage, by capillary
Active force reaches up to trapping nucleic acids area again on adsorptive pads.After waiting 2 minutes, extract work and complete.Then, U is dragged with handss
Type clips to amplification indicatrix, meanwhile, the insulating paper of battery case behind is taken out, after waiting 60 minutes, amplification is completed.And then, then
Secondary dragging is U-shaped to clip to detection indicatrix, is detected by capillary force, detects in the observation of test paper result observation window after 15 minutes
As a result.
In order to verify the feasibility of this device, extracted, expanded and detected using the Salmonella sample of variable concentrations,
As a result see Fig. 5, concentration is 106, 105, 104, 103, 102, 101(CFU/ml) sample control region and detection zone is red
Color, is judged to positive findingses, 100With NC control zones for it is red when, detection zone is feminine gender without color, result of determination.
In sum, whole process needs about 70min, simple to operate only needs 4 steps this utility model device when in use
(sample-adding → pressing extract control knob → dragging is U-shaped to clip to that amplification indicatrix → dragging is U-shaped to clip to reagent paper indicatrix), it is easy to take
Band.Compared with existing detection method, this device solves the problems, such as liquid storage, reduces cost and reduces complicated many more manipulations, is not required to
Can just be completed using large-scale instrument and technical professional.This utility model can be used for scarcity of resources area in future
To realize field quick detection.
Above content is only to illustrate technological thought of the present utility model, it is impossible to limit protection model of the present utility model with this
Enclose, it is every according to the utility model proposes technological thought, any change done on the basis of technical scheme each falls within this reality
Within protection domain with new claims.
Claims (6)
1. a kind of nucleic acid extraction, amplification and detection integrating device, it is characterised in that including the upper lid and pedestal that fasten up and down, base
Paper substrate nucleic acid extraction module, paper substrate nucleic acid isothermal amplification module and detection paper module are provided with seat；Use is offered on upper lid
Sample holes (1), control knob (2) and test paper result observation window (4) in Deca sample；In paper substrate nucleic acid isothermal amplification module
It is provided with the U-shaped folder (16) of movement, the side of upper lid is offered for fixing the fixing hole (3) of U-shaped folder (16)；U-shaped folder
(16), to drag end, the other end is paper for the sheet metal for being capable of heat conduction being closely superimposed including upper and lower two-layer, wherein one end
Section；The upper strata in scraps of paper area is inlaid with the scraps of paper (12) of the isothermal duplication reagent of the dependence unwindase of lyophilizing, the corresponding positions of lower floor
Put and be inlaid with nucleic acid absorption paper (11), drag end and stretched out by fixing hole (3)；
During nucleic acid extraction, the end of paper substrate cracking passage (8) in paper substrate nucleic acid extraction module is connected with nucleic acid absorption paper (11)
Touch, and the scraps of paper (12) are not contacted with nucleic acid absorption paper (11)；
During isothermal duplication, the heating plate (13) in paper substrate nucleic acid isothermal amplification module and nucleic acid absorption paper (11) and the scraps of paper (12) phase
Contact, is provided with the PMMA of Aerosol Pollution when avoiding nucleic acid amplification of sealing on nucleic acid absorption paper (11) and the scraps of paper (12)
Plate (15)；
During detection, test strips (18) and chromatographic solution holding tank (17) of the nucleic acid absorption paper (11) in detection paper module it
Between, and contact with the two.
2. nucleic acid extraction according to claim 1, amplification and detection integrating device, it is characterised in that the paper substrate nucleic acid
Extraction module includes cleanout fluid liquid storage tank (5) and lysate liquid storage tank (6), and lower end and the paper substrate of cleanout fluid liquid storage tank (5) are cleaned
The entrance of passage (9) is connected, and the entrance that the lower end of lysate liquid storage tank (6) cracks passage (8) with paper substrate is connected；Paper substrate cracking is logical
The lower section in road (8) arranges adsorptive pads (10)；The end of paper substrate flushing channel (9) arranges paper substrate valve, in cleaning, paper substrate valve
Contact with paper substrate cracking passage (8), paper substrate flushing channel (9) is connected with paper substrate cracking passage (8).
3. nucleic acid extraction according to claim 2, amplification and detection integrating device, it is characterised in that cleanout fluid storage
Caulking gum and sponge are provided with liquid pool (5) and lysate liquid storage tank (6), sponge is located at the lower section of caulking gum.
4. nucleic acid extraction according to claim 1, amplification and detection integrating device, it is characterised in that the paper substrate nucleic acid
Isothermal duplication module also includes the power supply (14) being connected with heating plate (13), and heating plate (13) adopts ceramic heating flake.
5. nucleic acid extraction according to claim 1, amplification and detection integrating device, it is characterised in that the detection paper
Module includes Sidestream chromatography test strips (18) and chromatographic solution holding tank (17), and chromatographic solution holding tank (17) is positioned at Sidestream chromatography paper slip
(18) below end；Sidestream chromatography paper slip (18) is corresponding with the position of the test paper result observation window (4) on upper lid.
6. nucleic acid extraction according to claim 5, amplification and detection integrating device, it is characterised in that the Sidestream chromatography
Paper slip (18) includes gum plate, sample pad, pad and the cellulose acetate membrane being cascading；Chromatographic solution holding tank
(17) it is placed with for storing the sponge of chromatographic solution.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201621127212.7U CN206127278U (en) | 2016-10-17 | 2016-10-17 | Nucleic acid extraction , increase and detection integrated device |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201621127212.7U CN206127278U (en) | 2016-10-17 | 2016-10-17 | Nucleic acid extraction , increase and detection integrated device |
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| Publication Number | Publication Date |
|---|---|
| CN206127278U true CN206127278U (en) | 2017-04-26 |
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ID=58573244
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| Application Number | Title | Priority Date | Filing Date |
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| CN201621127212.7U Active CN206127278U (en) | 2016-10-17 | 2016-10-17 | Nucleic acid extraction , increase and detection integrated device |
Country Status (1)
| Country | Link |
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| CN (1) | CN206127278U (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106520536A (en) * | 2016-10-17 | 2017-03-22 | 西安交通大学 | Nucleic-acid extraction, amplification and detection integrated device and manufacturing method and detection method thereof |
| CN110389057A (en) * | 2018-04-23 | 2019-10-29 | 上海伯顿医疗设备有限公司 | Integrated multi-functional biological sample processor |
| CN111879922A (en) * | 2020-07-13 | 2020-11-03 | 东南大学 | Integrated paper-based chip structure suitable for synchronous detection of nucleic acid and immunity and manufacturing method thereof |
-
2016
- 2016-10-17 CN CN201621127212.7U patent/CN206127278U/en active Active
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106520536A (en) * | 2016-10-17 | 2017-03-22 | 西安交通大学 | Nucleic-acid extraction, amplification and detection integrated device and manufacturing method and detection method thereof |
| CN110389057A (en) * | 2018-04-23 | 2019-10-29 | 上海伯顿医疗设备有限公司 | Integrated multi-functional biological sample processor |
| CN111879922A (en) * | 2020-07-13 | 2020-11-03 | 东南大学 | Integrated paper-based chip structure suitable for synchronous detection of nucleic acid and immunity and manufacturing method thereof |
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| TR01 | Transfer of patent right |
Effective date of registration: 20210514 Address after: 215129 South half of 2nd floor, building 3, 599 Taishan Road, high tech Zone, Suzhou City, Jiangsu Province Patentee after: Huicheng Zhilian Industrial Technology Service (Suzhou) Co.,Ltd. Address before: Beilin District Xianning West Road 710049, Shaanxi city of Xi'an province No. 28 Patentee before: XI'AN JIAOTONG University |
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Effective date of registration: 20211014 Address after: 215129 South half of floor 2, building 3, No. 599, Taishan Road, high tech Zone, Suzhou, Jiangsu Province Patentee after: Suzhou DianAn Biotechnology Co.,Ltd. Address before: 215129 South half of 2nd floor, building 3, 599 Taishan Road, high tech Zone, Suzhou City, Jiangsu Province Patentee before: Huicheng Zhilian Industrial Technology Service (Suzhou) Co.,Ltd. |