CN105277725B - A kind of integrated micro-flow control system for foranalysis of nucleic acids detection - Google Patents
A kind of integrated micro-flow control system for foranalysis of nucleic acids detection Download PDFInfo
- Publication number
- CN105277725B CN105277725B CN201410311068.1A CN201410311068A CN105277725B CN 105277725 B CN105277725 B CN 105277725B CN 201410311068 A CN201410311068 A CN 201410311068A CN 105277725 B CN105277725 B CN 105277725B
- Authority
- CN
- China
- Prior art keywords
- capillary electrophoresis
- nucleic acid
- module
- electrophoresis
- acid extraction
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
- 108020004707 nucleic acids Proteins 0.000 title claims abstract description 68
- 150000007523 nucleic acids Chemical class 0.000 title claims abstract description 68
- 102000039446 nucleic acids Human genes 0.000 title claims abstract description 68
- 238000001514 detection method Methods 0.000 title claims abstract description 23
- 238000005251 capillar electrophoresis Methods 0.000 claims abstract description 70
- 230000003321 amplification Effects 0.000 claims abstract description 45
- 238000003199 nucleic acid amplification method Methods 0.000 claims abstract description 45
- 238000000605 extraction Methods 0.000 claims abstract description 42
- 239000003153 chemical reaction reagent Substances 0.000 claims description 35
- 230000004888 barrier function Effects 0.000 claims description 30
- 238000001962 electrophoresis Methods 0.000 claims description 30
- 239000007788 liquid Substances 0.000 claims description 27
- 239000012530 fluid Substances 0.000 claims description 18
- 238000012546 transfer Methods 0.000 claims description 14
- 239000000758 substrate Substances 0.000 claims description 10
- 239000002699 waste material Substances 0.000 claims description 10
- 239000004926 polymethyl methacrylate Substances 0.000 claims description 8
- 239000003795 chemical substances by application Substances 0.000 claims description 7
- 229920003229 poly(methyl methacrylate) Polymers 0.000 claims description 7
- 239000011521 glass Substances 0.000 claims description 6
- 239000004205 dimethyl polysiloxane Substances 0.000 claims description 2
- 229920000435 poly(dimethylsiloxane) Polymers 0.000 claims description 2
- 235000013870 dimethyl polysiloxane Nutrition 0.000 claims 1
- CXQXSVUQTKDNFP-UHFFFAOYSA-N octamethyltrisiloxane Chemical compound C[Si](C)(C)O[Si](C)(C)O[Si](C)(C)C CXQXSVUQTKDNFP-UHFFFAOYSA-N 0.000 claims 1
- 238000004987 plasma desorption mass spectroscopy Methods 0.000 claims 1
- 238000000034 method Methods 0.000 abstract description 16
- 238000012360 testing method Methods 0.000 abstract description 8
- 238000004458 analytical method Methods 0.000 abstract description 3
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 6
- 230000000694 effects Effects 0.000 description 6
- 230000002068 genetic effect Effects 0.000 description 6
- 239000000463 material Substances 0.000 description 6
- ZHNUHDYFZUAESO-UHFFFAOYSA-N Formamide Chemical compound NC=O ZHNUHDYFZUAESO-UHFFFAOYSA-N 0.000 description 4
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 4
- 230000005611 electricity Effects 0.000 description 3
- 238000005516 engineering process Methods 0.000 description 3
- 239000000376 reactant Substances 0.000 description 3
- 239000000243 solution Substances 0.000 description 3
- 229910021642 ultra pure water Inorganic materials 0.000 description 3
- 239000012498 ultrapure water Substances 0.000 description 3
- 238000004804 winding Methods 0.000 description 3
- 240000007643 Phytolacca americana Species 0.000 description 2
- 239000003814 drug Substances 0.000 description 2
- 239000000835 fiber Substances 0.000 description 2
- 239000012634 fragment Substances 0.000 description 2
- 238000012252 genetic analysis Methods 0.000 description 2
- 238000007731 hot pressing Methods 0.000 description 2
- 239000012528 membrane Substances 0.000 description 2
- 239000011259 mixed solution Substances 0.000 description 2
- 229920003023 plastic Polymers 0.000 description 2
- 239000004033 plastic Substances 0.000 description 2
- 238000000197 pyrolysis Methods 0.000 description 2
- 230000009183 running Effects 0.000 description 2
- 238000007873 sieving Methods 0.000 description 2
- 230000009182 swimming Effects 0.000 description 2
- 208000026350 Inborn Genetic disease Diseases 0.000 description 1
- 229920005479 Lucite® Polymers 0.000 description 1
- 238000012408 PCR amplification Methods 0.000 description 1
- 206010036790 Productive cough Diseases 0.000 description 1
- 101710130181 Protochlorophyllide reductase A, chloroplastic Proteins 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 238000000429 assembly Methods 0.000 description 1
- 230000000712 assembly Effects 0.000 description 1
- 238000012742 biochemical analysis Methods 0.000 description 1
- 238000005842 biochemical reaction Methods 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 201000011510 cancer Diseases 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 238000010367 cloning Methods 0.000 description 1
- 238000004925 denaturation Methods 0.000 description 1
- 230000036425 denaturation Effects 0.000 description 1
- 238000013461 design Methods 0.000 description 1
- 238000003745 diagnosis Methods 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- XGZRAKBCYZIBKP-UHFFFAOYSA-L disodium;dihydroxide Chemical compound [OH-].[OH-].[Na+].[Na+] XGZRAKBCYZIBKP-UHFFFAOYSA-L 0.000 description 1
- 239000013536 elastomeric material Substances 0.000 description 1
- 239000005357 flat glass Substances 0.000 description 1
- 230000004907 flux Effects 0.000 description 1
- 238000004374 forensic analysis Methods 0.000 description 1
- 208000016361 genetic disease Diseases 0.000 description 1
- 238000010438 heat treatment Methods 0.000 description 1
- 239000012535 impurity Substances 0.000 description 1
- 238000001764 infiltration Methods 0.000 description 1
- 230000008595 infiltration Effects 0.000 description 1
- 238000007689 inspection Methods 0.000 description 1
- 238000012423 maintenance Methods 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 238000012856 packing Methods 0.000 description 1
- 229920002401 polyacrylamide Polymers 0.000 description 1
- -1 polydimethylsiloxane Polymers 0.000 description 1
- 238000003752 polymerase chain reaction Methods 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 239000011541 reaction mixture Substances 0.000 description 1
- 238000012216 screening Methods 0.000 description 1
- 238000007789 sealing Methods 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 210000003802 sputum Anatomy 0.000 description 1
- 208000024794 sputum Diseases 0.000 description 1
- 210000002700 urine Anatomy 0.000 description 1
Landscapes
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
- Apparatus Associated With Microorganisms And Enzymes (AREA)
Abstract
The invention discloses a kind of integrated micro-flow control system for foranalysis of nucleic acids detection.It includes nucleic acid extraction and amplification module and Capillary Electrophoresis module, is integrated in a microfluidic system to realize the full automation of foranalysis of nucleic acids testing process.The integrated microfluidic system of the present invention substantially reduces the volume and cost of hardware support kit equipment, staff is freed from the conventional operation flow process of numerous and diverse specialty.The use of this integrated system can provide more convenient, efficient, automatic, accurate, timely testing result for the application of many DNA such as clinical detection, forensic medical examination detection and analysis.
Description
Technical field
The present invention relates to a kind of integrated micro-flow control system for foranalysis of nucleic acids detection, belongs to biological detection analytical technology
Field.
Background technology
It is to examine for existing nucleic acid Capillary Electrophoresis based on complete or collected works' accepted way of doing sth foranalysis of nucleic acids detecting system of microflow control technique
The upgrading scheme that survey technology is proposed in the inferior positions such as automation, low cost and portability.Detection of nucleic acids is in many biochemical analysises
Field such as clinical medicine, forensic identification, heredity inspection etc. suffer from very great effect.On clinical medicine, genetic test
It is for the detection and diagnosis of various diseases, typical such as various malignant tumours, various ancestor genetic diseases；Lead in forensic analysis
Domain, genetic test are one of goldstandards for individual identification；In genetic analysis field, genetic test can be used to carry out parent-offspring
Identification, genetic map analysis etc..
Genetic test means used in every field are still traditional methods now, i.e., be manual extraction sample first
Nucleic acid in this, this step are usually to buy business-like nucleic acid extraction kit, are operated according still further to product description,
Carry out the biochemical reaction of series of complex to complete in test tube；PCR of the ensuing amplification flow process to commonly use
As a example by (PCR, Polymerase Chain Reaction) amplification, amplification is led to after needing to be manually adding to PCR reaction mixtures again
Cross commercial PCR instrument to carry out；Last detecting step is then needed by bulky HPCE to be detected
As a result.Therefore, if a system can be had, these steps above can be integrated and machine is transferred in all of operation
Automatically complete, artificial operation not only can be greatly reduced, evaded maloperation risk, and genetic test can be greatly improved
Analyze speed and detection flux.
The content of the invention
It is an object of the invention to provide a kind of integrated micro-flow control system for foranalysis of nucleic acids detection, the present invention can be by
Three step-nucleic acid extractions, nucleic acid amplification and Capillary Electrophoresis of traditional detection of nucleic acids-integrate and fully automatically complete
Into these steps, for a user, the present invention only requires user input sample to be tested (blood sample, urine sample, sputum sample, Fa Yijian
Timing is probably other more complicated samples) genetic analysis result is obtained.
Integrated micro-flow control system for foranalysis of nucleic acids detection provided by the present invention, including nucleic acid extraction and amplification mould
Block and Capillary Electrophoresis module；
With amplification module, the nucleic acid extraction includes that nucleic acid extracting reagent entrance on substrate, nucleic acid amplification agents enter
Mouth, sample entrance port and PCR chambers；The nucleic acid extracting reagent entrance, the sample entrance port and the PCR chambers are sequentially connected
It is logical, control valve for fluids I is provided between the sample entrance port and the PCR chambers, the arrival end of the PCR chambers is provided with filter paper；
The port of export of the PCR chambers is connected with control valve for fluids II, attachment structure and waste liquid outlet successively；The nucleic acid amplification examination
Agent entrance is connected with the fluid passage at the PCR chamber ingress end, and the nucleic acid amplification agents entrance is logical with the fluid
Control valve for fluids III is provided between road；
The Capillary Electrophoresis module includes the Capillary Electrophoresis sample channel on substrate and Capillary Electrophoresis supervisor
It is the setting that intersects vertically between road, and the Capillary Electrophoresis sample channel and the Capillary Electrophoresis main pipeline；The capillary
Pipe main pipeline is respectively arranged at two ends with Capillary Electrophoresis cathode can and Capillary Electrophoresis anode slot；The Capillary Electrophoresis sample feeding pipe
Road is provided with Capillary Electrophoresis waste liquid accumulator tank；
The attachment structure is included located at the PCR primer transfer chamber constituted on substrate of the nucleic acid extraction with amplification module
Room and electrophoresis related reagent storage chamber, lead between the PCR primer transfer chamber and the electrophoresis related reagent storage chamber
Cross 3 barrier films and be separated into independent chamber, barrier film A is located at the PCR located at the top of the PCR primer transfer chamber, barrier film B
Between product transfer chamber and the electrophoresis related reagent storage chamber, barrier film C is located at the electrophoresis related reagent storage chamber
Bottom；
The nucleic acid extraction is located at the top of the Capillary Electrophoresis module, and the electrophoresis related reagent with amplification module
Storage chamber is located at the hair located at the top of the sample introduction liquid storage tank of Capillary Electrophoresis, the sample introduction liquid storage tank of the Capillary Electrophoresis
It is connected in cons electrophoresis module and with the Capillary Electrophoresis sample channel.
In above-mentioned integrated micro-flow control system, the barrier film can be one layer of barrier film or laminated diaphragm.
In above-mentioned integrated micro-flow control system, the nucleic acid extraction with amplification module can by described in two panels substrate hot pressing key
Conjunction is obtained；
The Capillary Electrophoresis module can be obtained by two panels substrate thermocompression bonding.
In above-mentioned integrated micro-flow control system, the nucleic acid extraction with amplification module and the Capillary Electrophoresis module it
Between be provided with resilient middle layer.
In above-mentioned integrated micro-flow control system, the nucleic acid extraction is with amplification module specifically by polymethyl methacrylate
Make；
The Capillary Electrophoresis module is specifically made up of glass；
The resilient middle layer is specifically made up of PDMS.
In above-mentioned integrated micro-flow control system, the integrated micro-flow control system include 1～8 nucleic acid extraction with
Amplification module, such as 2 symmetrically arranged nucleic acid extractions and amplification module；
The Capillary Electrophoresis main pipeline of the Capillary Electrophoresis module 1～8, such as 2 symmetrically arranged capillary electricity
Swimming main pipeline；
Each described nucleic acid extraction and the attachment structure and 1 Capillary Electrophoresis main pipeline phase that expand in module
Connection.
Provided by the present invention for the integrated micro-flow control system of foranalysis of nucleic acids detection, can make traditional multiple with being arranged
Standby (PCR instrument, HPCE etc.) is integrated into an equipment, substantially reduces the volume and maintenance cost of equipment, makes gene
Detection is more efficient, instant, convenient, accurate.
Description of the drawings
Fig. 1 is overall top view of the present invention for the integrated micro-flow control system of foranalysis of nucleic acids detection.
Fig. 2 is the nucleic acid extraction and amplification module assembled axonometric drawing of device in Fig. 1.
Fig. 3 is the nucleic acid Capillary Electrophoresis module assembled axonometric drawing of device in Fig. 1.
Fig. 4 is the assembling axonometric drawing of two modules of device in Fig. 1.
Fig. 5 is the attachment structure schematic diagram of extraction amplification module and Capillary Electrophoresis module in the present invention.
Structural representations of the Fig. 6 for the attachment structure in Fig. 5 when in running order.
Fig. 7 is the integrated operation flow chart of the present invention.
Mark in figure is as follows：
I：Nucleic acid extraction and amplification module；II：Capillary Electrophoresis module；III：Resilient middle layer；I-U：Nucleic acid extraction with
The upper cover plate of amplification module；I-D：Nucleic acid extraction and the lower shoe for expanding module；II-U：The upper cover plate of Capillary Electrophoresis module；
II-D：The lower shoe of Capillary Electrophoresis module；
1st, nucleic acid extraction related reagent inflow entrance；2nd, nucleic acid amplification related reagent inflow entrance；3rd, sample adds mouth；4th, nucleic acid
Amplification related reagent control valve for fluids；5th, nucleic acid extraction related reagent control valve for fluids；6th, filter paper；7th, PCR chambers；8th, PCR is produced
Thing transfer control valve；9th, attachment structure；10th, waste liquid outlet；11st, cathode can；12nd, Capillary Electrophoresis waste liquid reservoir；13rd, anode
Groove；14th, Capillary Electrophoresis main pipeline；15th, Capillary Electrophoresis sample channel；16th, adding mouth diaphragm seal.9-A：Barrier film A；9-B：Every
Film B；9-C：Barrier film C；9-1：PCR primer transfer chamber；9-2：Electrophoresis related reagent storage chamber；9-II：Capillary Electrophoresis
Sample introduction liquid storage tank；9-III：Linked hole.
Specific embodiment
The present invention will be further described below in conjunction with the accompanying drawings, but the invention is not limited in following examples.
Absorption of the present embodiment using filter paper to DNA acts on extracting the DNA in sample, corresponding extraction phase with winding
Closing reagent has NaOH (NaOH) solution, dilute hydrochloric acid solution (HCl) and ultra-pure water.Side of the embodiment using PCR amplifications
Expanding target DNA fragments, the amplification related reagent for being used is pre- mixed PCR reactant liquors to formula.The embodiment is adopted and is compared
General right-angled intersection pipeline carrying out Capillary Electrophoresis, with the STR analysis (Short commonly used in judicial expertise
Tandem Repeat, STR) used as detection target, the electrophoresis related reagent for being used is formamide and commercialization STR scales by one
The mixed solution of certainty ratio.The following detailed description of the structure of the present invention.
Fig. 1 is a kind of feasible embodiment of the present invention.The device is integrally divided into two modules：Nucleic acid extraction and amplification
Module I and Capillary Electrophoresis module ii.Fig. 4 is the assembling mode of two modules：Nucleic acid extraction is placed in capillary with amplification module I
The top of electrophoresis module II, is embedded in one layer of resilient middle layer III between two modules.The use of resilient middle layer III is depended on
The material of two modules：If two modules are all rigid material (such as glass), resilient middle layer III can be connect with packing material
Microscopic gaps in contacting surface make two modules be fully contacted and good sealing function is played to junction；If a module
Jing is made (such as polydimethylsiloxane) using elastomeric material, then can not use resilient middle layer III.In the present embodiment
In, nucleic acid extraction with amplification module I material be PMMA (polymethyl methacrylate, lucite) plastics, Capillary Electrophoresis
The material of module ii is glass, and the material of resilient middle layer III is PDMS.
Fig. 2 is the package assembly of nucleic acid extraction and amplification module I, by two panels PMMA plastics I-U and I-D pass through hot pressing key and
Form.Three elastic membranes (PDMS) are embedded with two panels PMMA and form control valve for fluids 4,5 and 8, the structure of valve is to whole miniflow
Control system is operated and is controlled.In the present invention, valve type structure can also adopt mechanical ejector pin valve using traditional pneumatic operated valve
Even being other valve type structures, generally conventional valve type structure includes elastic membrane now, therefore must give in design
Consider.The valve type structure that the present embodiment is adopted is mechanical ejector pin valve, wherein mechanical thimble by supporting external equipment carry out operation with
Control, it is possible to achieve the closing of valve and opening function.Nucleic acid extraction and the upper cover plate I-U and nucleic acid extraction and amplification that expand module
One piece of filter paper 6 is inlaid with also in the middle part of two PMMA package assemblies of lower shoe I-D of module.The effect of filter paper 6 is to DNA using which
Winding effect the DNA in sample is dammed on filter paper 6, and other large granular impurities are (such as cell debris, various albumen
Deng) then can be flushed away through the fibre gap of filter paper 6.Filter paper 6 is located at the entrance side of PCR chambers 7, and itself falls within PCR
A part for chamber 7.The heater block of hardware support kit is located at the surface of PCR chambers 7.When the DNA in sample is wrapped in filter paper 6
After upper, the PCR amplifing reagents flowed into by nucleic acid amplification related reagent inflow entrance 2 can be full of PCR chambers 7, while will be filter paper 6 complete
Infiltration.During PCR, the template DNA on filter paper 6 can carry out reacting the amplification for completing DNA with the reactant liquor in PCR chambers 7
Process.After PCR reactions terminate, the PCR product in PCR chambers 7 is sucked up at the attachment structure 9 of upper and lower module.
The structure now of link block 9 is as shown in Figure 5.Which by three barrier film A9-A, barrier film B9-B and barrier film C9-C point is
Upper and lower two completely isolated PCR primer transfer chambers 9-1 and electrophoresis related reagent storage chamber 9-2.The wherein related examination of electrophoresis
The related reagent of electrophoresis is prestored in agent storage chamber 9-2, is for keeping the formamide of DNA denaturation and being used to identify here
The mixed solution of the commercialization scale of DNA length.PCR primer can be moved in PCR primer transfer chamber 9-1, due to barrier film B9-
The effect of B, which is entirely isolated with electrophoresis reagents.Linked hole on the lower section of barrier film C9-C as resilient middle layer III
9-III.Linked hole 9-III is to be seamlessly connected with the sample introduction liquid storage tank 9-II on Capillary Electrophoresis module ii, can be considered as one
Individual entirety.Electrophoresis pipeline is inoculated with the screening of electrophoresis in (including Capillary Electrophoresis main pipeline 14 and Capillary Electrophoresis sample channel 15)
Medium (used here as linear polyacrylamide LPA), sieving media can be full of the liquid storage tank 9-II and linked hole of Capillary Electrophoresis
9-III.But due to the effect of barrier film C9-C, LPA and electrophoresis reagents are also to separate completely.When PCR primer is full of connection knot
During PCR primer transfer chamber 9-1 in structure 9, the sample introduction electrode IV on hardware support kit will be operated, and move downward, will be every
Film A9-A, barrier film B9-B, barrier film C9-C are all poked, and now the state of attachment structure 9 is as shown in Figure 6.By barrier film A9-A, barrier film
After B9-B, barrier film C9-C are poked, PCR primer transfer chamber 9-1, electrophoresis related reagent storage chamber 9-2, linked hole 9-III with
And sample introduction liquid storage tank 9-II is just fully coupled together.So, PCR primer just can be fully with electrophoresis related reagent
Mix and down as sample introduction electrode IV touches sieving media LPA.At this time, as long as adding on sample introduction electrode IV
Upper voltage, it is possible to enter the flow process of electrophoretic separation detection.
Assembling axonometric drawings of the Fig. 3 for Capillary Electrophoresis module, this module are made by glass, used DNA
The commonplace right-angled intersection pipeline of chip capillary cataphoresis.Electrophoresis is etched on the upper cover plate II-U of Capillary Electrophoresis module
Pipeline (Capillary Electrophoresis main pipeline 14 and Capillary Electrophoresis sample channel 15) and carve out Capillary Electrophoresis liquid storage tank 9-II,
Waste liquid liquid storage tank 12, cathode can 11 and anode slot 13, afterwards with another glass as Capillary Electrophoresis module lower shoe II-
D, two sheet glass are combined in the way of heating key sum.After this device is fixed on its hardware support kit, waste liquid storage
Iontophoretic electrode in liquid pool 12, cathode can 11 and anode slot 13 can be inserted in corresponding liquid storage tank and keep fixed.But capillary
The electrode IV of the sample introduction liquid storage tank 9-II of electrophoresis tube can be just operated when being only moved at attachment structure 9 to PCR primer, poke every
Film A9-A, barrier film B9-B, barrier film C9-C are simultaneously inserted in the liquid storage tank 9-II of Capillary Electrophoresis.
Integrated operation flow charts of the Fig. 7 for apparatus of the present invention.First whole device is assembled, will nucleic acid extraction and expansion
Increase module I, Capillary Electrophoresis module ii and resilient middle layer III to stack according to shown in Fig. 3.Afterwards by the device for stacking
It is installed in corresponding hardware support kit equipment, hardware device should be able to be by nucleic acid extraction and amplification module I and capillary electricity
Swimming module ii is firmly compressed.So whole device is just completed.Then we enter into the operating process shown in Fig. 7.To adopt
The sample for collecting is added in sample addition mouth 3, and is thoroughly sealed adding mouth with adding mouth diaphragm seal 16.The two steps are
Only manual steps in whole DNA detection process, remaining work are completely given supporting hardware device and are carried out.
According to shown in Fig. 7, hardware device can open control valve for fluids 4 and 8 first, close control valve for fluids 5.It is pre-stored in hard
Cell pyrolysis liquid (being NaOH NaOH here) on part equipment can be sucked up to sample from nucleic acid extraction related reagent inflow entrance 1
This addition mouth 3, makes the cell in sample fully crack.Afterwards, watery hydrochloric acid can be passed through from nucleic acid extraction related reagent inflow entrance 1 molten
In liquid and cell pyrolysis liquid DNA solution is pumped to into PCR chambers 7, in aspiration procedure, the winding effect of filter paper 6 is by template
DNA is firmly captured in its fibre gap.Then, nucleic acid extraction related reagent inflow entrance 1 can be passed through ultra-pure water to the miscellaneous of residual
Matter (such as cell fragment, various albumen etc.) is cleaned, and waste liquid is flowed out from waste liquid outlet 10.After ultra-pure water is cleaned, DNA's carries
Take flow process to terminate, control valve for fluids 4 cuts out, control valve for fluids 5 and 8 is opened, PCR reactant liquors are flowed into by nucleic acid amplification related reagent
Mouth 2 is sucked up to PCR chambers 7, simultaneously closes off control valve for fluids 4,5 and 8 afterwards, completely encloses PCR chambers 7.Now, temperature
Degree circulation comes into operation into performing PCR and reacts.After question response terminates, PCR primer is pumped to into attachment structure 9.Now, DNA cloning
Flow process terminates.After PCR primer full of after attachment structure upper chamber 9-1, electrophoresis sample introduction electrode IV runnings, poke barrier film A9-A, every
Film B9-B and barrier film C9-C, makes PCR primer be sufficiently mixed with electrophoresis reagents.Meanwhile, other electricity on Capillary Electrophoresis module ii
Pole is also simultaneously upper electric, PCR primer is separated and is detected according to classical chip capillary cataphoresis flow process.Detection and voltage control
System is completed by hardware support kit, and after the completion of detection, direct output result, completes Aulomatizeted Detect flow process.
According to this flow process, the process of DNA detections will become very efficient, automatic and convenient.
Claims (6)
1. it is a kind of for foranalysis of nucleic acids detection integrated micro-flow control system, it is characterised in that：It includes nucleic acid extraction with amplification
Module and Capillary Electrophoresis module；
The nucleic acid extraction and amplification module include nucleic acid extracting reagent entrance on substrate, nucleic acid amplification agents entrance,
Sample entrance port and PCR chambers；The nucleic acid extracting reagent entrance, the sample entrance port and the PCR chambers are sequentially connected logical, institute
State and between sample entrance port and the PCR chambers, be provided with control valve for fluids I, the arrival end of the PCR chambers is provided with filter paper；It is described
The port of export of PCR chambers is connected with control valve for fluids II, attachment structure and waste liquid outlet successively；The nucleic acid amplification agents enter
Mouthful be connected with the fluid passage at the PCR chamber ingress end, and the nucleic acid amplification agents entrance and the fluid passage it
Between be provided with control valve for fluids III；
The Capillary Electrophoresis module includes Capillary Electrophoresis sample channel and Capillary Electrophoresis main pipeline on substrate, and
It is the setting that intersects vertically between the Capillary Electrophoresis sample channel and the Capillary Electrophoresis main pipeline；The capillary supervisor
Road is respectively arranged at two ends with Capillary Electrophoresis cathode can and Capillary Electrophoresis anode slot；Set on the Capillary Electrophoresis sample channel
There is Capillary Electrophoresis waste liquid accumulator tank；
The attachment structure include located at the PCR primer transfer chamber constituted on the substrate of the nucleic acid extraction with amplification module and
Electrophoresis related reagent storage chamber, passes through 3 between the PCR primer transfer chamber and the electrophoresis related reagent storage chamber
Barrier film is separated into independent chamber, and barrier film A is located at the PCR primer located at the top of the PCR primer transfer chamber, barrier film B
Between transfer chamber and the electrophoresis related reagent storage chamber, barrier film C is under the electrophoresis related reagent storage chamber
Portion；
The nucleic acid extraction is stored located at the top of the Capillary Electrophoresis module, and the electrophoresis related reagent with amplification module
Chamber is located at the hair located at the top of the sample introduction liquid storage tank of Capillary Electrophoresis, the sample introduction liquid storage tank of the Capillary Electrophoresis
It is connected in cons electrophoresis module and with the Capillary Electrophoresis sample channel.
2. integrated micro-flow control system according to claim 1, it is characterised in that：The barrier film is one layer of barrier film or multilayer
Barrier film.
3. integrated micro-flow control system according to claim 1 and 2, it is characterised in that：The nucleic acid extraction and amplification mould
Block is obtained by two panels substrate thermocompression bonding；
The Capillary Electrophoresis module is obtained by two panels substrate thermocompression bonding.
4. integrated micro-flow control system according to claim 3, it is characterised in that：The nucleic acid extraction with amplification module with
Resilient middle layer is provided between the Capillary Electrophoresis module.
5. integrated micro-flow control system according to claim 4, it is characterised in that：The nucleic acid extraction with amplification module by
Polymethyl methacrylate is made；
The Capillary Electrophoresis module is made up of glass；
The resilient middle layer is made up of PDMS.
6. integrated micro-flow control system according to claim 5, it is characterised in that：The integrated micro-flow control system includes
1～8 nucleic acid extraction and amplification module；
The Capillary Electrophoresis module includes 1～8 Capillary Electrophoresis main pipeline；
Each described nucleic acid extraction is connected with 1 Capillary Electrophoresis main pipeline with the attachment structure in amplification module
It is logical.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410311068.1A CN105277725B (en) | 2014-07-01 | 2014-07-01 | A kind of integrated micro-flow control system for foranalysis of nucleic acids detection |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410311068.1A CN105277725B (en) | 2014-07-01 | 2014-07-01 | A kind of integrated micro-flow control system for foranalysis of nucleic acids detection |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN105277725A CN105277725A (en) | 2016-01-27 |
| CN105277725B true CN105277725B (en) | 2017-03-29 |
Family
ID=55147093
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201410311068.1A Active CN105277725B (en) | 2014-07-01 | 2014-07-01 | A kind of integrated micro-flow control system for foranalysis of nucleic acids detection |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN105277725B (en) |
Families Citing this family (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US11135592B2 (en) | 2013-06-27 | 2021-10-05 | Quark Biosciences Taiwan, Inc. | Multiplex slide plate device having storage tank |
| CN105296348A (en) * | 2015-11-20 | 2016-02-03 | 融智生物科技（青岛）有限公司 | Genotyping detection-based microfluidic chip, detection system and device |
| CN107099598B (en) * | 2017-05-24 | 2020-05-08 | 济南市疾病预防控制中心 | Microfluidic integrated detection method for bacteria |
| CN107460121A (en) * | 2017-07-22 | 2017-12-12 | 新疆昆泰锐生物技术有限公司 | A kind of PCR reaction systems are prepared and sampling device and full-automatic PCR instrument |
| CN108728327B (en) * | 2018-05-23 | 2021-11-23 | 博奥生物集团有限公司 | Full-automatic sample preparation micro-fluidic system and preparation method and application method thereof |
| CN109622083A (en) * | 2019-01-24 | 2019-04-16 | 京东方科技集团股份有限公司 | A kind of microfluidic system and preparation method thereof |
| CN112442435A (en) * | 2019-08-28 | 2021-03-05 | 宁波海尔施基因科技有限公司 | Clamping device integrating PCR amplification and capillary electrophoresis analysis and detection method |
| TW202214866A (en) * | 2020-09-30 | 2022-04-16 | 富佳生技股份有限公司 | Heating structure, detection chip, nucleic acid detection disc and nucleic acid detection device |
| CN114317222A (en) * | 2020-09-30 | 2022-04-12 | 富佳生技股份有限公司 | Nucleic acid detecting cassette and nucleic acid detecting apparatus |
| TW202214868A (en) * | 2020-09-30 | 2022-04-16 | 富佳生技股份有限公司 | Nucleic acid detection disc and nucleic acid detection device |
| CN113702476A (en) * | 2021-08-24 | 2021-11-26 | 聚光科技(杭州)股份有限公司 | Sample injection chip and method based on electrophoresis technology |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1973197A (en) * | 2004-01-28 | 2007-05-30 | 挪其普公司 | A diagnostic system for carrying out a nucleic acid sequence amplification and detection process |
| CN102239005A (en) * | 2009-11-03 | 2011-11-09 | 天津微纳芯科技有限公司 | Integrated detection chip and application method thereof |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| NZ580374A (en) * | 2007-04-04 | 2012-06-29 | Netbio Inc | Plastic microfluidic separation and detection platforms |
-
2014
- 2014-07-01 CN CN201410311068.1A patent/CN105277725B/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1973197A (en) * | 2004-01-28 | 2007-05-30 | 挪其普公司 | A diagnostic system for carrying out a nucleic acid sequence amplification and detection process |
| CN102239005A (en) * | 2009-11-03 | 2011-11-09 | 天津微纳芯科技有限公司 | Integrated detection chip and application method thereof |
Non-Patent Citations (2)
| Title |
|---|
| 《A Simple, Valveless Microfluidic Sample Preparation Device for Extraction and Amplification of DNA from Nanoliter-Volume Samples》;Lindsay A. Legendre et.al;《Analytical Chemistry》;20060301;第78卷(第5期);第1444-1451页 * |
| 《microfluidic device for DNA sequencing：sample preparation and electrophoresis analysis》;Brian M Paegel et.al;《Current Opinion in Biotechnology》;20131231;第42-50页 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN105277725A (en) | 2016-01-27 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN105277725B (en) | A kind of integrated micro-flow control system for foranalysis of nucleic acids detection | |
| CN204727866U (en) | A kind ofly collect the micro fluidic device that nucleic acid amplification and microarray are detected on one | |
| US8895292B2 (en) | Microfluidic chip devices and their use | |
| CN106520536B (en) | Nucleic acid extraction, amplification and detection integrating device and its manufacture method and detection method | |
| Esfandyarpour et al. | Gate-controlled microfluidic chamber with magnetic bead for DNA sequencing-by-synthesis technology | |
| CN112538425A (en) | On-chip nucleic acid amplification detection system and method based on micro-fluidic chip | |
| Jayamohan et al. | Advances in microfluidics and lab-on-a-chip technologies | |
| KR20230031971A (en) | Portable nucleic acid analysis system and high-performance microfluidic electroactive polymer actuators | |
| CN107502544B (en) | Micro-fluidic chip detection control system | |
| CN105296348A (en) | Genotyping detection-based microfluidic chip, detection system and device | |
| CN209923319U (en) | Microfluidic chip structure and analysis device | |
| CN107430138A (en) | The instrument of diagnostic test is performed for fluid cartridge | |
| CN100537219C (en) | Methods and apparatus for pathogen detection and analysis | |
| Zhuang et al. | Fully automated sample preparation microsystem for genetic testing of hereditary hearing loss using two-color multiplex allele-specific PCR | |
| CN111394234B (en) | Digital chip and method for nucleic acid amplification | |
| CN104360091B (en) | A kind of chip, using method and purposes | |
| CN207276626U (en) | A kind of micro-fluidic chip detecting and controlling system | |
| CN108414522A (en) | The device of all-in-one machine is detected for micro-fluidic chip and the more flux of NC films | |
| CN205152235U (en) | A micro -fluidic chip , detecting system and device for gene somatotype detects | |
| CN111289762A (en) | Micro-fluidic chip sample adding device and testing method | |
| CN103484353A (en) | Biomacromolecule extracting device based on filter paper | |
| WO2022174470A1 (en) | Fully integrated pathogen nucleic acid analyzer | |
| CN102998472B (en) | Automatic device for heterogeneous immunization rapid analysis and use method of automatic device | |
| CN211826105U (en) | Micro-fluidic chip application of sample device | |
| CN102162807B (en) | Nucleic acid array analysis platform and method based on microfluidic chip |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |