CN205152235U - A micro -fluidic chip , detecting system and device for gene somatotype detects - Google Patents
A micro -fluidic chip , detecting system and device for gene somatotype detects Download PDFInfo
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- CN205152235U CN205152235U CN201520934070.4U CN201520934070U CN205152235U CN 205152235 U CN205152235 U CN 205152235U CN 201520934070 U CN201520934070 U CN 201520934070U CN 205152235 U CN205152235 U CN 205152235U
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Abstract
The utility model discloses a micro -fluidic chip that the gene somatotype detected, this chip include stock solution unit, at least one nucleic acid extraction unit, at least one increase of nucleic acid PCR unit, at least one electrophoresis unit and fluid the control unit. The utility model discloses gene somatotype detecting system and gene somatotype detection device including an above -mentioned micro -fluidic chip are further disclosed. Adopt technical scheme can realize that gene somatotype quick, high -efficient, automatic, the high flux detects, is favorable to this technique further to promote and popularize.
Description
Technical field
The utility model relates to biological medicine instrument, particularly relates to a kind of micro-fluidic chip for genotype tests, and comprises genotype tests system and the proofing unit of this chip.
Background technology
Genotyping technique, with the detection sensitivity of its outstanding bio-identification resolving power and intimate trace, has been widely used in the fields such as animals and plants species identification, nosophyte numerator somatotype, people's identification (Forensic Identification, paternity test etc.), disease related gene type analysis.At present, genotype tests process based on electrophoretic technique comprise DNA extraction with quantitatively, multiplexed PCR amplification and three key steps such as electrophoretic separation and detection, the multiple different instrument of routine analysis requirements of process and reaction system, major part step and reaction system between transfer all relate to manual operations, and generally can only have been come by professional in specialized laboratory, time-consuming, effort, expensive.The microminiaturization developing into genotype tests of microflow control technique, automatization and facilitation provide effective way, as the monotechnicss such as efficient chip DNA extraction, fast chip pcr amplification, high-throughput microchip electrophoresis and integrated in achieved obvious progress, there is a collection of work had their own characteristics each, embodied that micro-fluidic chip volume is little, sample consumption less, the feature such as analysis cost is low, speed of response is fast.
Although quick for the micro-fluidic chip researchdevelopment of gene type, but existing method majority only can complete one or two step in genotype tests flow process, still need to consider the problem such as to be connected with normal experiment flow process, not remarkable to the improvement of whole analysis process, limit its practical application.In recent years, successively there is the full automatic genotype tests instrument of a few money, as RapidHIT
tM200, MiDAS system, DNAScan etc.These products are all core with microflow control technique, by operate with conventional fluid and automatic technology combines, nucleic acid extraction, amplification and electrophoresis are incorporated on an instrument and complete.But these systems mostly are part and utilize microflow control technique, do not give full play to the advantage of micro-total analysis system, thus cause that detection system level of automation is not high, poor reliability, complex structure, with high costs, be unfavorable for large-scale promotion and application.
Therefore, need to provide a kind of novel detection method, to meet along with user improves the requirement of analysis platform, day by day to genotyping system microminiaturization, automatization, fast and efficiently demand.
Utility model content
The technical problems to be solved in the utility model is to provide a kind of micro-fluidic chip for genotype tests, and comprise genotype tests system and the proofing unit of this chip, to solve, genotype tests system automation degree in prior art is not high, poor reliability, complex structure, the problem such as with high costs.
For solving the problems of the technologies described above, the utility model adopts following technical proposals:
For a micro-fluidic chip for genotype tests, this chip comprises
Liquid storage unit, for storing in testing process the liquid using or produce;
At least one nucleic acid extraction unit, based on micro-structure surface modification, extracts the nucleic acid substances in sample fluid;
At least one nucleic acid PCR amplification unit, based on the nucleic acid substances obtained, carries out pcr amplification, obtains PCR primer;
At least one electrophoretic cell, carries out electrophoretic analysis to pcr amplification product, obtains PCR separated product;
Flow control unit, based on the mode that micro-extruding promotes, controls the fluid direction of travel in testing process.
Preferably, described liquid storage unit comprises nucleic acid extraction buffer liquid storage tank, sample reservoir, washing fluid liquid storage tank, elutriant liquid storage tank, PCR reaction solution liquid storage tank, electrophoresis sample inlet pool, electrophoretic buffer liquid storage tank and waste liquid pool;
Described nucleic acid extraction buffer liquid storage tank, sample reservoir, washing fluid liquid storage tank are communicated with described nucleic acid extraction unit by microchannel with elutriant liquid storage tank;
Described PCR reaction solution liquid storage tank is communicated with pcr amplification unit with described nucleic acid extraction unit by microchannel with electrophoresis sample inlet pool.
Preferably, the degree of depth of described microchannel is 20 ~ 60 μm.
Preferably, described nucleic acid extraction unit adopts the fluid channel of the micro-pillar array had through surface modification treatment.
Preferably, the fluid channel described in micro-pillar array is " S " type fluid channel.
Preferably, the fluid channel of described micro-pillar array deposits the thick silicon oxide of 50 ~ 100nm.
Preferably, described flow control unit adopts Integrated microfluidic assembly to carry out fluid control.
Preferably, described microfluidic components is the micro-valve group of Elastic Film or Elastic Film Micropump group.
Preferably, described microfluidic components comprises the first miniflow control, the second miniflow control, the 3rd miniflow control, the 4th miniflow control, the 5th miniflow control, the 6th miniflow control, the 7th miniflow control, the 8th miniflow control, the 9th miniflow control, the tenth miniflow control and the 11 miniflow control;
Described first miniflow control is connected with nucleic acid extraction buffer liquid storage tank and sample reservoir respectively;
Described second miniflow control and the 3rd miniflow control are successively set between the first miniflow control and nucleic acid extraction unit;
Described 6th miniflow control is connected with nucleic acid extraction unit, nucleic acid PCR amplification unit and electrophoresis sample inlet pool respectively;
Described 4th miniflow control and the 5th miniflow control are successively set between PCR reaction solution liquid storage tank and the 6th miniflow control;
Described 7th miniflow control is connected with nucleic acid PCR amplification unit and waste liquid pool respectively;
Described 8th miniflow control is connected with the second miniflow control, the 9th miniflow control and washing fluid liquid storage tank respectively;
Described 9th miniflow control is connected with the second miniflow control, the 8th miniflow control and elutriant liquid storage tank respectively;
Described tenth miniflow control is connected with waste liquid pool, nucleic acid extraction unit and the 6th miniflow control respectively;
Described 11 miniflow control is connected with waste liquid pool, nucleic acid PCR amplification unit and the 7th miniflow control respectively.
Preferably, described nucleic acid extraction unit, nucleic acid PCR amplification unit and electrophoretic cell are respectively two.
Preferably, described sample reservoir, PCR reaction solution liquid storage tank and microchip electrophoresis enter night pond is two groups.
Comprise a genotype tests system for micro-fluidic chip described above, this system comprises
Micro-fluidic chip as above;
For providing the power control unit of extruding power for flow control unit;
For providing the temperature control unit of temperature of reaction for pcr amplification unit;
For providing the voltage control unit of response voltage for electrophoretic cell; With
For carrying out the fluorescence detection unit of fluorimetric analysis to PCR separated product.
Comprise a genotype tests device for micro-fluidic chip described above, this device comprises many groups of being arranged on carrier micro-fluidic chip as above.
Preferably, described many group micro-fluidic chip annular arrays are arranged on the carrier.
Preferably, described many group micro-fluidic chips share an electrophoretic buffer liquid storage tank.
Preferably, this device comprises further
The temperature control unit of temperature of reaction is provided for the pcr amplification unit for organizing micro-fluidic chip more;
The voltage control unit of response voltage is provided for the electrophoretic cell for organizing micro-fluidic chip more; With
For carrying out the fluorescence detection unit of fluorimetric analysis for the PCR separated product of many group micro-fluidic chips.
The beneficial effects of the utility model are as follows:
Technical scheme advantage described in the utility model is:
1, using with micro-pillar array and through the microchannel of surface modification as nucleic acid extraction unit, this structure can adopt the micro fabrication of standard to make, effectively can improve controllability and the stability of the course of processing, thus significantly improve the reproducibility and reliability of method for extracting nucleic acid;
2, the nucleic acid extraction unit based on Micro Channel Architecture can adopt the integrated valve/Micropump that declines as flow control unit, the elementary cell be convenient to the gene types such as nucleic acid extraction, pcr amplification, electrophoretic separation and detection operate is integrated in same chip, and be easy to the multiple sample of parallel analysis, thus effectively improve integrated level and the level of automation of chip system.
Accompanying drawing explanation
Below in conjunction with accompanying drawing, embodiment of the present utility model is described in further detail;
Fig. 1 illustrates the schematic diagram of micro-fluidic chip described in the utility model;
Fig. 2 illustrates the schematic diagram of liquid storage unit described in micro-fluidic chip described in the utility model;
Fig. 3 illustrates the schematic diagram of flow control unit described in micro-fluidic chip described in the utility model;
Fig. 4 illustrates the schematic diagram of genotype tests system described in the utility model;
Fig. 5 illustrates the schematic diagram of genotype tests device described in the utility model;
Drawing reference numeral
The nucleic acid extraction unit of the 1-1: the first sample, the nucleic acid extraction unit of the 1-2: the second sample, the nucleic acid PCR amplification unit of the 2-1: the first sample, the nucleic acid PCR amplification unit of the 2-2: the second sample, 3-1: the first sample amplification product microchip electrophoresis unit, the 3-2: the second sample amplification product microchip electrophoresis unit;
R1: nucleic acid extraction buffer liquid storage tank, the liquid storage tank of the R2: the first sample, the liquid storage tank of the R3: the second sample, R4: washing fluid liquid storage tank, R5: elutriant liquid storage tank, the PCR reaction solution liquid storage tank of the R6-1: the first sample, the PCR reaction solution liquid storage tank of the R6-2: the second sample, the microchip electrophoresis sample inlet pool of the R8-1: the first sample, the microchip electrophoresis sample inlet pool of the R8-2: the second sample, R9: waste liquid pool, R10: electrophoretic buffer liquid storage tank, R11: waste liquid pool, R12: electrophoretic buffer liquid storage tank;
The miniflow control label of the first sample: V1-1, V3-1, V5-1, V6-1, V7-1, V8-1, V11-1;
The miniflow control label of the second sample: V1-2, V3-2, V5-2, V6-2, V7-2, V8-2, V11-2;
Share miniflow control label: V2, V4, V9, V10.
Embodiment
In order to be illustrated more clearly in the utility model, below in conjunction with preferred embodiments and drawings, the utility model is described further.Parts similar in accompanying drawing represent with identical Reference numeral.It will be appreciated by those skilled in the art that specifically described content is illustrative and nonrestrictive below, protection domain of the present utility model should do not limited with this.
The utility model discloses a kind of micro-fluidic chip for genotype tests, and comprise genotype tests system and the proofing unit of this chip.Micro-fluidic chip described in this programme adopts the microchannel containing surface modification micro-pillar array as nucleic acid extraction unit, and adopt integrated form film micro-valve/Micropump that the elementary cell that the gene types such as lysis/DNA extraction, DNA cloning, electrophoretic separation and detection operate is integrated in same chip, automatization, fast and efficiently genotype tests can be realized.
As shown in Figure 1, concrete the utility model discloses a kind of micro-fluidic chip for genotype tests, this micro-fluidic chip comprises the liquid storage unit for storing in testing process the liquid using or produce, at least one is based on modification, extract the nucleic acid extraction unit of the nucleic acid substances in sample fluid, at least one is based on the nucleic acid substances obtained, carry out pcr amplification, obtain the nucleic acid PCR amplification unit of PCR primer, at least one carries out electrophoretic analysis to pcr amplification product, acquisition PCR primer is separated, the electrophoretic cell detected and analyze and the mode promoted based on micro-extruding, to the flow control unit that the fluid direction of travel in testing process controls.
As shown in Figure 2, described liquid storage unit comprises nucleic acid extraction buffer liquid storage tank, sample reservoir, washing fluid liquid storage tank, elutriant liquid storage tank, PCR reaction solution liquid storage tank, electrophoresis sample inlet pool, electrophoretic buffer liquid storage tank and waste liquid pool; Described nucleic acid extraction buffer liquid storage tank, sample reservoir, washing fluid liquid storage tank are communicated with described nucleic acid extraction unit by microchannel with elutriant liquid storage tank; Described PCR reaction solution liquid storage tank is communicated with pcr amplification unit with described nucleic acid extraction unit by microchannel with electrophoresis sample inlet pool; Described electrophoretic buffer liquid storage tank is communicated with described electrophoretic cell by microchannel; Described waste liquid pool is communicated with nucleic acid PCR amplification unit with described nucleic acid extraction unit by microchannel.In this programme, described nucleic acid extraction unit adopts the fluid channel of the micro-pillar array had through surface modification treatment, described in there is micro-pillar array fluid channel be " S " type fluid channel.
Described flow control unit adopts the Integrated microfluidic assembly of the micro-valve group of such as Elastic Film or Elastic Film Micropump group to carry out fluid control; Wherein, described microfluidic components comprises the first miniflow control, the second miniflow control, the 3rd miniflow control, the 4th miniflow control, the 5th miniflow control, the 6th miniflow control, the 7th miniflow control, the 8th miniflow control, the 9th miniflow control, the tenth miniflow control and the 11 miniflow control; Described first miniflow control is connected with nucleic acid extraction buffer liquid storage tank and sample reservoir respectively; Described second miniflow control and the 3rd miniflow control are successively set between the first miniflow control and nucleic acid extraction unit; Described 6th miniflow control is connected with nucleic acid extraction unit, nucleic acid PCR amplification unit and electrophoresis sample inlet pool respectively; Described 4th miniflow control and the 5th miniflow control are successively set between PCR reaction solution liquid storage tank and the 6th miniflow control; Described 7th miniflow control is connected with nucleic acid PCR amplification unit and waste liquid pool respectively; Described 8th miniflow control is connected with the second miniflow control, the 9th miniflow control and washing fluid liquid storage tank respectively; Described 9th miniflow control is connected with the second miniflow control, the 8th miniflow control and elutriant liquid storage tank respectively; Described tenth miniflow control is connected with waste liquid pool, nucleic acid extraction unit and the 6th miniflow control respectively; Described 11 miniflow control is connected with waste liquid pool, nucleic acid PCR amplification unit and the 7th miniflow control respectively.
In this programme, this optimum system choosing by nucleic acid extraction unit described in two groups, nucleic acid PCR amplification unit and electrophoretic cell combine with the micro-valve group of Elastic Film or Elastic Film Micropump group, formation can carry out one group of micro-fluidic chip of Dual channel detection simultaneously, in order to coordinate the detection of two groups of samples in this group micro-fluidic chip, be respectively each sample and be equipped with sample reservoir, PCR reaction solution liquid storage tank and microchip electrophoresis enter pond at night, and nucleic acid extraction buffer liquid storage tank, washing fluid liquid storage tank, elutriant liquid storage tank, waste liquid pool and electrophoretic buffer liquid storage tank can adopt same group, to save the use of chip space and material.
As shown in Figure 4, the utility model further discloses a kind of genotype tests system comprising micro-fluidic chip described above, this system comprise micro-fluidic chip as above, for provide for flow control unit extruding power power control unit, for providing the temperature control unit of temperature of reaction for pcr amplification unit, for providing the voltage control unit of response voltage and the fluorescence detection unit for carrying out fluorimetric analysis to PCR separated product for electrophoretic cell.Wherein, in order to coordinate the micro-valve group of Elastic Film or Elastic Film Micropump group to carry out liquid-flow control to the sample liquid in chip, power control unit described in this programme adopts the orifice plate corresponding with valve/pump point each in chip, air feeder and controls by liquid flow direction the confession airgun controller that air feeder is each valve/pump point air feed.In this programme, described temperature control unit adopts controlling with the use of to the temperature of reaction of pcr amplification unit of temperature sensor, heating unit and temperature regulator.In this programme, the power-supply controller of electric that described voltage control unit comprises the electrode be arranged in electrophoretic buffer liquid storage tank, provides for electrode the high-voltage power supply of high voltage electric and control power supply to export, with the separation making electrophoretic cell complete PCR primer under hyperbaric environment.In this programme, in order to complete the detection of gene type further, being equipped with the fluorescence detection unit with multicolor fluorescence agent in systems in which, fluoroscopic examination being carried out to PCR separated product, obtains final detection analytical results.
The utility model further discloses a kind of genotype tests device, and this device comprises many groups of being arranged on carrier micro-fluidic chip as above; This many groups micro-fluidic chip annular array is arranged on to be had on the carrier of regular geometric shapes.In this programme, preferably 12 above-mentioned micro-fluidic chip annular arrays are arranged on circular carrier.In order to save use operating key, in this programme, many group micro-fluidic chips adopt a running buffer liquid pool to carry out PCR primer separation.This device comprises further provides the power control unit of extruding power for the flow control unit for organizing micro-fluidic chip more, provide the temperature control unit of temperature of reaction for the pcr amplification unit for organizing micro-fluidic chip, provide the voltage control unit of response voltage and the fluorescence detection unit for carrying out fluorimetric analysis for the PCR separated product of many group micro-fluidic chips for the electrophoretic cell for organizing micro-fluidic chip more more.
As shown in Figures 2 and 3, the workflow of micro-fluidic chip described in the utility model: first, carries out preparation load procedure: add sample 1 and sample 2 respectively in two groups of sample reservoir, add corresponding reagent respectively in other liquid storage tanks, then, enter sample introduction mixing step: order opens micro-valve V1-1 and V1-2, micro-valve V3-1 and V3-2, micro-valve V5-1 and V5-2 and V9, after nucleic acid extraction buffer in nucleic acid extraction buffer liquid storage tank R1 is mixed with sample 1 and sample 2 respectively, enter in nucleic acid extraction unit, DNA molecular in sample 1 and sample 2 is adsorbed on mini column array structure 1-1 and mini column array structure 1-2 surface respectively, then enters waste liquid pool R7 with the nucleic acid extraction buffer waste liquid crossed, then, enter rinse step: order opens micro-valve V2, micro-valve V3-1 and V3-2, micro-valve V5-1 and V5-2 and V9, the dcq buffer liquid in dcq buffer liquid liquid storage tank R4 is made to flow through mini column array structure 1-1 and mini column array structure 1-2 in two nucleic acid extraction unit respectively, to remove the impurity of mini column array structure surface adsorption, the dcq buffer liquid used subsequently enters waste liquid pool R7, then, enter elution step: order opens micro-valve V4, micro-valve V3-1 and V3-2, micro-valve V5-1 and V5-2 and V10, elution buffer in elution buffer liquid storage tank R5 is made to enter mini column array structure 1-1 and mini column array structure 1-2 in two nucleic acid extraction unit respectively, the DNA molecular of channel surface is adsorbed on wash-out, meanwhile, order opens micro-valve V6-1 and V6-2, micro-valve V7-1 and V7-2, micro-valve V8-1 and V8-2 and V10, after PCR reaction solution in PCR reaction solution liquid storage tank R6-1 and PCR reaction solution liquid storage tank R6-2 is mixed with the DNA profiling molecule 1 eluted and DNA profiling molecule 2 respectively, enter pcr amplification unit, the mixing solutions of PCR reaction solution and elutriant be filled with pcr amplification unit after remaining solution enter waste liquid pool R9, then, enter pcr amplification step: close all micro-valves, chip pcr amplification is carried out to mixed DNA profiling molecule 1 and DNA profiling molecule 2, finally, enter electrophoretic analysis step: after pcr amplification terminates, open micro-valve V11-1 and V11-2, sample introduction voltage is applied to electrophoresis sample inlet pool R8-1 with between electrophoresis sample inlet pool R8-2 and waste liquid pool R11, pcr amplification product is made to enter electrophoresis sample intake passage, this sample intake passage is the sample introduction part of electrophoretic cell, with split tunnel square crossing, subsequently, between the electrophoretic buffer liquid storage tank R10 and electrophoretic buffer liquid storage tank R12 at electrophoretic separation passage 3-1 and electrophoretic separation passage two ends, apply separation voltage, make PCR primer realize being separated, determination and analysis.
Below by one group of embodiment, the utility model is described further:
As shown in Figures 1 to 3, in the present embodiment, this micro-fluidic chip comprises three parts: lysis and DNA extraction unit, pcr amplification unit, electrophoretic separation unit, adopt the integrated valve/Micropump that declines to be connected with microchannel between various piece, genotype tests process that is quick, efficient, automatization can be completed.Wherein, nucleic acid extraction unit adopts the microchannel containing surface modification micro-pillar array as the stationary phase of nucleic acid extraction, this structure can adopt the micro fabrication of standard to make, such one side improves the reproducibility and reliability that DNA extraction unit makes, also enhance on the other hand the compatibility of itself and integrated form Micropump/micro-valve flow control unit, thus effectively improve integrated level and the level of automation of chip.
As shown in Figure 4, above-mentioned micro-fluidic chip with for provide for flow control unit extrude power power control unit, for providing the temperature control unit of temperature of reaction for pcr amplification unit, for providing the voltage control unit of response voltage and the fluorescence detection unit common constitutive gene somatotype detection system for carrying out fluorimetric analysis to PCR separated product for electrophoretic cell, by the genotype tests of the complete two groups of samples of cooperation of the sample path on micro-fluidic chip and each control unit.In this example, described power control unit adopts the orifice plate corresponding with valve/pump point each in chip, air feeder and controls by liquid flow direction the confession airgun controller that air feeder is each valve/pump point air feed.Described temperature control unit adopts controlling with the use of to the temperature of reaction of pcr amplification unit of temperature sensor, heating unit and temperature regulator.The power-supply controller of electric that described voltage control unit comprises the electrode be arranged in electrophoretic buffer liquid storage tank, provides for electrode the high-voltage power supply of high voltage electric and control power supply to export, with the separation making electrophoretic cell complete PCR primer under hyperbaric environment.Also be equipped with the fluorescence detection unit with multicolor fluorescence agent in system, fluoroscopic examination carried out to PCR separated product, obtains final detection analytical results.
As shown in Figure 5, a kind of genotype tests device is provided further in this example, this device comprises annular array and is arranged on 12 groups of micro-fluidic chips as above on round carrier, the power control unit of extruding power is provided for the flow control unit for organizing micro-fluidic chip more, the temperature control unit of temperature of reaction is provided for the pcr amplification unit for organizing micro-fluidic chip more, there is provided the voltage control unit of response voltage and the fluorescence detection unit for carrying out fluorimetric analysis for the PCR separated product of many group micro-fluidic chips for the electrophoretic cell for organizing micro-fluidic chip more.Often organize micro-fluidic chip can detect two samples simultaneously, utilize the array chip apparatus described in this example to detect 24 samples simultaneously, significantly improve level of automation and sample throughput compared with existing methods of genotyping, realize the mass of genotype tests, automatization, rapid detection and analysis.In this example, many group micro-fluidic chips adopt a running buffer liquid pool to carry out PCR primer separation.This device comprises further.
Described in this example, micro-fluidic chip is for glass-chip, first designs chip channel configuration and makes mask, then etches required microchannel according to the glass photomask/wet-etching technology of standard, and its etching depth is 40 μm; The substrate ultrasonic drilling device etched punches in corresponding position; After cleaning, the microchannel surface cvd silicon oxide of the micro-pillar array in nucleic acid extraction unit as the stationary phase of nucleic acid extraction, the thickness about 50 ~ 100nm of this silicon oxide, other parts masking film; Surface treated substrate is aimed at the blank cover plate of same size and is put into vacuum drying oven successively and fits and temperature programmed control stove thermal bonding, can obtain required glass-chip.Before using, adopt polydimethylsiloxanefilm film (PDMS), its thickness is about 150-250 μm, and be placed in the middle of the gentle circuit control layer of glass-chip and carry out reversible sealing-in, can complete micro-fluidic chip.
In sum, technical scheme described in the utility model is using with micro-pillar array and through the microchannel of surface modification as nucleic acid extraction unit, this structure can adopt the micro fabrication of standard to make, effectively can improve controllability and the stability of the course of processing, thus significantly improve the reproducibility and reliability of method for extracting nucleic acid; The program can adopt the integrated valve/Micropump that declines as flow control unit based on the nucleic acid extraction unit of Micro Channel Architecture, the elementary cell be convenient to the gene types such as nucleic acid extraction, pcr amplification, electrophoretic separation and detection operate is integrated in same chip, and be easy to the multiple sample of parallel analysis, thus effectively improve integrated level and the level of automation of chip system.
Obviously; above-described embodiment of the present utility model is only for the utility model example is clearly described; and be not the restriction to embodiment of the present utility model; for those of ordinary skill in the field; can also make other changes in different forms on the basis of the above description; here cannot give exhaustive to all embodiments, every belong to the technical solution of the utility model the apparent change of extending out or variation be still in the row of protection domain of the present utility model.
Claims (16)
1. for a micro-fluidic chip for genotype tests, it is characterized in that, this chip comprises
Liquid storage unit, for storing in testing process the liquid using or produce;
At least one nucleic acid extraction unit, based on micro-structure surface modification, extracts the nucleic acid substances in sample fluid;
At least one nucleic acid PCR amplification unit, based on the nucleic acid substances obtained, carries out pcr amplification, obtains PCR primer;
At least one electrophoretic cell, carries out electrophoretic analysis to pcr amplification product, obtains PCR separated product;
Flow control unit, based on the mode that micro-extruding promotes, controls the fluid direction of travel in testing process.
2. micro-fluidic chip according to claim 1, it is characterized in that, described liquid storage unit comprises nucleic acid extraction buffer liquid storage tank, sample reservoir, washing fluid liquid storage tank, elutriant liquid storage tank, PCR reaction solution liquid storage tank, electrophoresis sample inlet pool, electrophoretic buffer liquid storage tank and waste liquid pool;
Described nucleic acid extraction buffer liquid storage tank, sample reservoir, washing fluid liquid storage tank are communicated with described nucleic acid extraction unit by microchannel with elutriant liquid storage tank;
Described PCR reaction solution liquid storage tank is communicated with pcr amplification unit with described nucleic acid extraction unit by microchannel with electrophoresis sample inlet pool;
Described electrophoretic buffer liquid storage tank is communicated with described electrophoretic cell by microchannel;
Described waste liquid pool is communicated with nucleic acid PCR amplification unit with described nucleic acid extraction unit by microchannel.
3. micro-fluidic chip according to claim 2, is characterized in that, the degree of depth of described microchannel is 20 ~ 60 μm.
4. micro-fluidic chip according to claim 1, is characterized in that, described nucleic acid extraction unit adopts the fluid channel of the micro-pillar array had through surface modification treatment.
5. micro-fluidic chip according to claim 4, is characterized in that, described in there is micro-pillar array fluid channel be " S " type fluid channel.
6. micro-fluidic chip according to claim 5, is characterized in that, the fluid channel of described micro-pillar array deposits the silicon oxide that 50 ~ 100nm is thick.
7. micro-fluidic chip according to claim 2, is characterized in that, described flow control unit adopts Integrated microfluidic assembly to carry out fluid control.
8. micro-fluidic chip according to claim 7, is characterized in that, described microfluidic components is the micro-valve group of Elastic Film or Elastic Film Micropump group.
9. micro-fluidic chip according to claim 7, it is characterized in that, described microfluidic components comprises the first miniflow control, the second miniflow control, the 3rd miniflow control, the 4th miniflow control, the 5th miniflow control, the 6th miniflow control, the 7th miniflow control, the 8th miniflow control, the 9th miniflow control, the tenth miniflow control and the 11 miniflow control;
Described first miniflow control is connected with nucleic acid extraction buffer liquid storage tank and sample reservoir respectively;
Described second miniflow control and the 3rd miniflow control are successively set between the first miniflow control and nucleic acid extraction unit;
Described 6th miniflow control is connected with nucleic acid extraction unit, nucleic acid PCR amplification unit and electrophoresis sample inlet pool respectively;
Described 4th miniflow control and the 5th miniflow control are successively set between PCR reaction solution liquid storage tank and the 6th miniflow control;
Described 7th miniflow control is connected with nucleic acid PCR amplification unit and waste liquid pool respectively;
Described 8th miniflow control is connected with the second miniflow control, the 9th miniflow control and washing fluid liquid storage tank respectively;
Described 9th miniflow control is connected with the second miniflow control, the 8th miniflow control and elutriant liquid storage tank respectively;
Described tenth miniflow control is connected with waste liquid pool, nucleic acid extraction unit and the 6th miniflow control respectively;
Described 11 miniflow control is connected with waste liquid pool, nucleic acid PCR amplification unit and the 7th miniflow control respectively.
10. micro-fluidic chip according to claim 8, is characterized in that, described nucleic acid extraction unit, nucleic acid PCR amplification unit and electrophoretic cell are respectively two.
11. micro-fluidic chips according to claim 10, is characterized in that, it is two groups that described sample reservoir, PCR reaction solution liquid storage tank and microchip electrophoresis enter night pond.
12. 1 kinds of genotype tests systems comprising micro-fluidic chip as claimed in claim 1, it is characterized in that, this system comprises
Micro-fluidic chip as claimed in claim 1;
For providing the power control unit of extruding power for flow control unit;
For providing the temperature control unit of temperature of reaction for pcr amplification unit;
For providing the voltage control unit of response voltage for electrophoretic cell; With
For carrying out the fluorescence detection unit of fluorimetric analysis to PCR separated product.
13. 1 kinds of genotype tests devices comprising micro-fluidic chip as claimed in claim 1, it is characterized in that, this device comprises many groups of being arranged on carrier micro-fluidic chip as claimed in claim 1.
14. genotype tests devices according to claim 13, is characterized in that, described many group micro-fluidic chip annular arrays are arranged on the carrier.
15. genotype tests devices according to claim 13, is characterized in that, described many group micro-fluidic chips share an electrophoretic buffer liquid storage tank.
16. genotype tests devices according to claim 13, it is characterized in that, this device comprises further
The power control unit of extruding power is provided for the flow control unit for organizing micro-fluidic chip more;
The temperature control unit of temperature of reaction is provided for the pcr amplification unit for organizing micro-fluidic chip more;
The voltage control unit of response voltage is provided for the electrophoretic cell for organizing micro-fluidic chip more; With
For carrying out the fluorescence detection unit of fluorimetric analysis for the PCR separated product of many group micro-fluidic chips.
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Cited By (4)
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| 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 |
| CN108118088A (en) * | 2016-11-28 | 2018-06-05 | 中国科学院大连化学物理研究所 | A kind of people's Y-STR gene loci classifying methods based on micro-fluidic chip |
| CN109839417A (en) * | 2017-11-28 | 2019-06-04 | 中国科学院大连化学物理研究所 | A kind of huamn autosomal str locus site classifying method based on micro-fluidic chip |
| WO2020078410A1 (en) * | 2018-10-17 | 2020-04-23 | 北京致雨生物科技有限公司 | Sample treatment device and method, and digital pcr system comprising treatment device |
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2015
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| 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 |
| CN108118088A (en) * | 2016-11-28 | 2018-06-05 | 中国科学院大连化学物理研究所 | A kind of people's Y-STR gene loci classifying methods based on micro-fluidic chip |
| CN108118088B (en) * | 2016-11-28 | 2021-08-27 | 中国科学院大连化学物理研究所 | Human Y-STR gene locus typing method based on micro-fluidic chip |
| CN109839417A (en) * | 2017-11-28 | 2019-06-04 | 中国科学院大连化学物理研究所 | A kind of huamn autosomal str locus site classifying method based on micro-fluidic chip |
| CN109839417B (en) * | 2017-11-28 | 2021-08-27 | 中国科学院大连化学物理研究所 | Human autosomal STR (short tandem repeat) gene locus typing method based on micro-fluidic chip |
| WO2020078410A1 (en) * | 2018-10-17 | 2020-04-23 | 北京致雨生物科技有限公司 | Sample treatment device and method, and digital pcr system comprising treatment device |
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