CN110358681A - A kind of portable PCR device - Google Patents
A kind of portable PCR device Download PDFInfo
- Publication number
- CN110358681A CN110358681A CN201910614406.1A CN201910614406A CN110358681A CN 110358681 A CN110358681 A CN 110358681A CN 201910614406 A CN201910614406 A CN 201910614406A CN 110358681 A CN110358681 A CN 110358681A
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- pcr
- reaction
- pcr device
- chip
- module
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- 238000006243 chemical reaction Methods 0.000 claims abstract description 172
- 239000000523 sample Substances 0.000 claims description 27
- 238000001514 detection method Methods 0.000 claims description 26
- 238000010438 heat treatment Methods 0.000 claims description 24
- 238000012545 processing Methods 0.000 claims description 18
- 238000007789 sealing Methods 0.000 claims description 16
- NJPPVKZQTLUDBO-UHFFFAOYSA-N novaluron Chemical compound C1=C(Cl)C(OC(F)(F)C(OC(F)(F)F)F)=CC=C1NC(=O)NC(=O)C1=C(F)C=CC=C1F NJPPVKZQTLUDBO-UHFFFAOYSA-N 0.000 claims description 14
- 239000000463 material Substances 0.000 claims description 13
- 238000004458 analytical method Methods 0.000 claims description 12
- 238000001816 cooling Methods 0.000 claims description 11
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 claims description 9
- GPRLSGONYQIRFK-UHFFFAOYSA-N hydron Chemical compound [H+] GPRLSGONYQIRFK-UHFFFAOYSA-N 0.000 claims description 8
- 238000009434 installation Methods 0.000 claims description 8
- 229910052799 carbon Inorganic materials 0.000 claims description 6
- 230000005669 field effect Effects 0.000 claims description 4
- -1 phosphate anion Chemical class 0.000 claims description 4
- 229910019142 PO4 Inorganic materials 0.000 claims description 3
- XPPKVPWEQAFLFU-UHFFFAOYSA-J diphosphate(4-) Chemical compound [O-]P([O-])(=O)OP([O-])([O-])=O XPPKVPWEQAFLFU-UHFFFAOYSA-J 0.000 claims description 3
- 229910021389 graphene Inorganic materials 0.000 claims description 3
- 150000002500 ions Chemical group 0.000 claims description 3
- CWQXQMHSOZUFJS-UHFFFAOYSA-N molybdenum disulfide Chemical compound S=[Mo]=S CWQXQMHSOZUFJS-UHFFFAOYSA-N 0.000 claims description 3
- 229910052982 molybdenum disulfide Inorganic materials 0.000 claims description 3
- 239000002070 nanowire Substances 0.000 claims description 3
- 239000010452 phosphate Substances 0.000 claims description 3
- 239000013078 crystal Substances 0.000 claims description 2
- 238000003825 pressing Methods 0.000 claims description 2
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- 238000001727 in vivo Methods 0.000 claims 1
- 230000035484 reaction time Effects 0.000 abstract description 5
- 238000012360 testing method Methods 0.000 abstract description 4
- 239000000243 solution Substances 0.000 description 19
- 238000000034 method Methods 0.000 description 6
- 239000004065 semiconductor Substances 0.000 description 6
- 238000012408 PCR amplification Methods 0.000 description 5
- 238000002347 injection Methods 0.000 description 5
- 239000007924 injection Substances 0.000 description 5
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 5
- 238000000018 DNA microarray Methods 0.000 description 3
- 238000010586 diagram Methods 0.000 description 3
- 238000005516 engineering process Methods 0.000 description 3
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 2
- GWEVSGVZZGPLCZ-UHFFFAOYSA-N Titan oxide Chemical compound O=[Ti]=O GWEVSGVZZGPLCZ-UHFFFAOYSA-N 0.000 description 2
- 230000003321 amplification Effects 0.000 description 2
- 238000007405 data analysis Methods 0.000 description 2
- 238000000354 decomposition reaction Methods 0.000 description 2
- 235000013399 edible fruits Nutrition 0.000 description 2
- 238000013467 fragmentation Methods 0.000 description 2
- 238000006062 fragmentation reaction Methods 0.000 description 2
- 230000001939 inductive effect Effects 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 238000003199 nucleic acid amplification method Methods 0.000 description 2
- 230000035945 sensitivity Effects 0.000 description 2
- PBCFLUZVCVVTBY-UHFFFAOYSA-N tantalum pentoxide Inorganic materials O=[Ta](=O)O[Ta](=O)=O PBCFLUZVCVVTBY-UHFFFAOYSA-N 0.000 description 2
- WYTZZXDRDKSJID-UHFFFAOYSA-N (3-aminopropyl)triethoxysilane Chemical compound CCO[Si](OCC)(OCC)CCCN WYTZZXDRDKSJID-UHFFFAOYSA-N 0.000 description 1
- 102000016928 DNA-directed DNA polymerase Human genes 0.000 description 1
- 108010014303 DNA-directed DNA polymerase Proteins 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- PNEYBMLMFCGWSK-UHFFFAOYSA-N aluminium oxide Inorganic materials [O-2].[O-2].[O-2].[Al+3].[Al+3] PNEYBMLMFCGWSK-UHFFFAOYSA-N 0.000 description 1
- 229910000147 aluminium phosphate Inorganic materials 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 238000003759 clinical diagnosis Methods 0.000 description 1
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- 239000004205 dimethyl polysiloxane Substances 0.000 description 1
- 235000013870 dimethyl polysiloxane Nutrition 0.000 description 1
- 238000009826 distribution Methods 0.000 description 1
- 230000005518 electrochemistry Effects 0.000 description 1
- 238000005265 energy consumption Methods 0.000 description 1
- 238000007667 floating Methods 0.000 description 1
- 238000000799 fluorescence microscopy Methods 0.000 description 1
- 239000012634 fragment Substances 0.000 description 1
- CJNBYAVZURUTKZ-UHFFFAOYSA-N hafnium(IV) oxide Inorganic materials O=[Hf]=O CJNBYAVZURUTKZ-UHFFFAOYSA-N 0.000 description 1
- 238000002955 isolation Methods 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 229910052751 metal Inorganic materials 0.000 description 1
- 239000002184 metal Substances 0.000 description 1
- 239000011259 mixed solution Substances 0.000 description 1
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- 238000012986 modification Methods 0.000 description 1
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- 239000003068 molecular probe Substances 0.000 description 1
- 239000002159 nanocrystal Substances 0.000 description 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 description 1
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 description 1
- 229940085991 phosphate ion Drugs 0.000 description 1
- NBIIXXVUZAFLBC-UHFFFAOYSA-N phosphoric acid Substances OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 description 1
- 238000004987 plasma desorption mass spectroscopy Methods 0.000 description 1
- 229920000435 poly(dimethylsiloxane) Polymers 0.000 description 1
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- 229910001845 yogo sapphire Inorganic materials 0.000 description 1
Classifications
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
- B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
- B01L3/508—Containers for the purpose of retaining a material to be analysed, e.g. test tubes rigid containers not provided for above
- B01L3/5085—Containers for the purpose of retaining a material to be analysed, e.g. test tubes rigid containers not provided for above for multiple samples, e.g. microtitration plates
- B01L3/50851—Containers for the purpose of retaining a material to be analysed, e.g. test tubes rigid containers not provided for above for multiple samples, e.g. microtitration plates specially adapted for heating or cooling samples
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L7/00—Heating or cooling apparatus; Heat insulating devices
- B01L7/52—Heating or cooling apparatus; Heat insulating devices with provision for submitting samples to a predetermined sequence of different temperatures, e.g. for treating nucleic acid samples
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2200/00—Solutions for specific problems relating to chemical or physical laboratory apparatus
- B01L2200/14—Process control and prevention of errors
- B01L2200/143—Quality control, feedback systems
- B01L2200/147—Employing temperature sensors
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2300/00—Additional constructional details
- B01L2300/18—Means for temperature control
- B01L2300/1805—Conductive heating, heat from thermostatted solids is conducted to receptacles, e.g. heating plates, blocks
Landscapes
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Clinical Laboratory Science (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Life Sciences & Earth Sciences (AREA)
- Biochemistry (AREA)
- Molecular Biology (AREA)
- Analytical Chemistry (AREA)
- Hematology (AREA)
- Apparatus Associated With Microorganisms And Enzymes (AREA)
Abstract
The present invention relates to a kind of portable PCR device, including for the power module of power supply, shell, setting operation on the housing and display module, be arranged in the shell and reaction module with microwell array formula PCR reaction chip and be arranged in the shell and for providing the temperature control execution module of reaction temperature to the PCR reaction on the PCR reaction chip.PCR device of the invention uses microwell array formula PCR reaction chip, it is not required to external droplet generator, the single template samples being diluted can be directly dispersed in micropore set on PCR reaction chip, and it is powered by power module to the PCR device, it realizes the PCR reaction of sample by PCR reaction chip and temperature control execution module and PCR is reacted and reaction temperature is provided, the setting and display to parameters such as the reaction time of the PCR device, temperature curve and cycle-indexes are realized by operation and display module, structure is simple, easy to operate, testing cost is cheap.
Description
Technical field
The invention belongs to technical field of molecular biology, and in particular to a kind of portable PCR device.
Background technique
Polymerase chain reaction, abbreviation PCR are that a kind of Protocols in Molecular Biology is for amplifying specific DNA fragmentation
One of most common Protocols in Molecular Biology.
Droplet type PCR device is needed before amplification as current common one of PCR device to the core after Macrodilution
Acid sample solution carries out droplet processing, single template samples to be dispersed in the microreactor or droplet of chip；And to sample
This dropletization need to use droplet generator, and present droplet generator common configuration is complicated, and size is larger, causes to be adapted to therewith
PCR device it is not portable and at high price.
Currently, the semiconductor biochip technology with microwell array is quickly grown, it is right in biology, chemical analysis
It detects sample size and reagent consumption is all very low, therefore, the semiconductor biochip technology with microwell array is in molecular biosciences
It learns, there is good development prospect in chemistry or clinical diagnosis.It is based on as Chinese patent CN107083326A discloses one kind
The microchamber array digital pcr chip of PDMS material, which disclose the chip structures comprising injection port, microchamber array
And outlet, the injection port include sample feeding mouth and water injection port, the outlet includes that sample outlet and water go out sample
Mouthful；The water injection port is connected to the water microchamber array positioned at chip periphery with water outlet；The sample feeding mouth and sample go out
Sample mouth is connected to each reaction chamber in microchamber array by microchannel.The chip can be used for PCR reaction.
However, at present there has been no document report by the semiconductor biochip applications of microwell array in the PCR in PCR reaction
The concrete structure design of device.
Summary of the invention
A kind of portable PCR device is provided the invention aims to overcome the deficiencies in the prior art.
In order to achieve the above objectives, the technical scheme adopted by the invention is as follows:
A kind of portable PCR device, the PCR device include power module, shell, setting for power supply in the shell
Operation and display module on body, be arranged in the shell and the reaction module with microwell array formula PCR reaction chip and
It is arranged in the shell and for providing the temperature control execution module of reaction temperature to the PCR reaction on the PCR reaction chip.
Further, the reaction module includes for installing the hinge assembly of the PCR reaction chip, being arranged described
The intracorporal circuit board of shell and the reaction controlling unit being arranged on the circuit board, the hinge assembly are arranged in the circuit board
Top, the reaction controlling unit are electrically connected to control the reaction that the temperature control execution module provides with the temperature control execution module
Temperature.
Further, the PCR device further includes for sealing the micropore on the PCR reaction chip and making each micro-
Mutually isolated sealing element between hole, the hinge assembly include being arranged on the circuit board and being used to install the PCR
The hinge pedestal of reaction chip and one end and the rotation connection of one end of the hinge pedestal and for the sealing element to be compressed
Hinge lid on the PCR reaction chip.
In terms of an implementation according to the present invention, the hinge assembly further include be arranged in the hinge lid the other end and
For with the hinge pedestal clamping or isolated hook.
According to an embodiment of the invention, the hinge pedestal include base body and be arranged on the base body and
For installing the chip mounting blocks of the PCR reaction chip, the hinge lid and the base body are rotatablely connected, the hook
It is clamped or separates with the base body.
According to an embodiment of the invention, the hinge lid includes the conjunction of one end rotation connection of one end and the hinge pedestal
Page lid ontology and the chip briquetting for being arranged on the hinge lid ontology and be used to be pressed on the PCR reaction chip, the extension
The other end of the hinge lid ontology is arranged in hook.
Further, the temperature control execution module includes being arranged below the circuit board and reacting core with the PCR
Heat-sink unit below the heating unit and with the heating unit thermal contact conductance is arranged in the heating unit of piece thermal contact conductance
And the temperature sensor for detecting the reaction temperature, the heating unit, heat-sink unit and temperature sensor respectively with institute
State the electrical connection of reaction controlling unit.
It is further preferred that the heating unit includes heat-conducting block, is arranged below the heat-conducting block and leads with described
The heater of heat block thermal contact conductance and the thermally conductive block pressur plate being arranged in above the heater, the heat-conducting block are located at described thermally conductive
Between the block pressur plate and heater and upper end of the heat-conducting block sequentially passes through the thermally conductive block pressur plate and circuit board and institute respectively
State the contact of PCR reaction chip.
In terms of certain specific implementations, the upper surface of the heat-conducting block opens up groove, and the temperature sensor is arranged in institute
It states in groove.Preferably, the temperature sensor is film temperature sensor.
It is further preferred that carbon film is arranged between the contact surface of both the PCR reaction chip and described heating unit.
It is further preferred that the heat-sink unit include be arranged below the heating unit and with the heating unit
The cooling fin of thermal contact conductance, be arranged in below the cooling fin and with the thermally conductive cover of the cooling fin thermal contact conductance and setting exist
Fan in the thermally conductive cover.
In terms of certain implementations according to the present invention, the PCR device further includes at the detection being arranged on the circuit board
Unit is managed, the micropore bottom of the PCR reaction chip is provided with biochemical sensor, and the detection processing unit is described for acquiring
The signal of biochemical sensor simultaneously carries out processing analysis to signal, then feeds back analysis result to the operation and display module.
Certain some embodiment according to the present invention, the surface of the biochemical sensor is coated with hydrogen ion sensitive material, phosphoric acid
Radical ion or pyrophosphate ion sensitive material.
Certain other embodiment according to the present invention, the biochemical sensor are ion-sensitive field effect transistor, nanometer
One of line, graphene or crystal of molybdenum disulfide tube sensor or miniature electrochemical.
Some embodiments according to the present invention, the hinge pedestal are equipped with and are formed with the biochemical sensor pin pad
The probe of electrical connection, the probe are connect with the detection processing unit to acquire the electric signal of the biochemical sensor.
Further, the detection processing unit includes data acquisition circuit, ADC chip and processor, and the data are adopted
Collector acquires the signal of micropore bottom biochemical sensor, after carrying out analog-to-digital conversion by ADC chip, is conveyed to processor and is counted
Word signal processing is simultaneously analyzed, and then feeds back analysis result to the operation and display module.
Further, 0.1 ten thousand~10,000,000 micropores, each micropore are contained in the microwell array of the PCR reaction chip
Volume be 1fL~10pL.
Further, the shell include upper end have opening enclosure body and with the enclosure body rotation connection and
For covering the case top lid for closing the opening, the operation and display module setting cover on the housing, the PCR device
Further include the support plate for being arranged in the enclosure body and being connect with the enclosure body upper end, is provided in the support plate
For installing the installation position of the reaction module, the reaction module is arranged on the installation position, the temperature control execution module
It is arranged below the reaction module.
Further, the PCR device further includes tweezers and vacuum pencil, is provided in the support plate and is respectively used to put
The tweezers for setting the tweezers and vacuum pencil place position and vacuum pencil placement position.
Further, the power module includes being arranged in the intracorporal power panel of the shell, is separately positioned on the shell
On electric source line interface and power switch.USB interface is also set up on the housing.
Further, the operation and display module include that setting touch display screen curtain on the housing and setting exist
Screen control plate between the touch display screen curtain and shell.
Due to the application of the above technical scheme, compared with the prior art, the invention has the following advantages:
PCR device of the invention use microwell array formula PCR reaction chip, be not required to external droplet generator, can directly by
Reaction solution containing the single template samples being diluted is dispersed in micropore set on PCR reaction chip, and passes through power module
It powers to the PCR device, the PCR reaction of sample is realized by PCR reaction chip and temperature control execution module and offer is reacted to PCR
Reaction temperature is realized by operation and display module to parameters such as the reaction time of the PCR device, temperature curve and cycle-indexes
Setting and display, easy to operate, testing cost is cheap.
PCR device of the invention is not required to external droplet generator, and reaction system is significantly reduced, and realizes that instrument is small
Type.And structure is simple, light weight, and easy to carry, instrument price is cheap.
Detailed description of the invention
Fig. 1 is the stereoscopic schematic diagram of the portable PCR device of one embodiment of the present of invention；
Fig. 2 is the schematic rear view of the portable PCR device of one embodiment of the present of invention；
Fig. 3 is the elevational schematic view of the portable PCR device of one embodiment of the present of invention；
Fig. 4 is the schematic cross-sectional view of the portable PCR device of one embodiment of the present of invention；
Fig. 5 is the decomposition diagram of the portable PCR device of one embodiment of the present of invention；
Fig. 6 is that the decomposition of the hinge assembly and PCR reaction chip of the portable PCR device of one embodiment of the present of invention is shown
It is intended to；
Fig. 7 is the PCR reaction knot that the PCR device of one embodiment of invention is used for a specific PCR reaction chip
The two-dimentional heating power schematic diagram of number when the detection of fruit；
Fig. 8 is the PCR reaction knot that the PCR device of one embodiment of invention is used for a specific PCR reaction chip
Sensor signal histogram histogram representative when the detection of fruit；
In figure: 1, PCR reaction chip；2, hinge assembly；201, base body；202, hinge lid ontology；203, chip is pacified
Fill block；204, chip mounting groove；205, chip briquetting；206, it links up with；3, circuit board；4, heat-conducting block；5, heater；6, heat-conducting block
Pressing plate；7, temperature sensor；8, cooling fin；9, fan；10, thermally conductive cover；11, enclosure body；1101, shell side plate；1101a,
Shell side plate body part；1101b, interface interconnecting piece；1102, housing base；12, case top lid；13, support plate；14, power supply is opened
It closes；15, power panel；16, touch display screen curtain；17, screen control plate；18, tweezers；19, vacuum pencil.
Specific embodiment
With reference to the accompanying drawings of the specification and specific embodiment is further described through the present invention.
Referring to shown in Fig. 1~6, the portable PCR device of one embodiment of the present of invention, including the power supply mould for power supply
Block, shell, the operation being arranged on shell and display module, setting are in shell and have microwell array formula PCR reaction chip 1
Reaction module and be arranged in shell and the temperature control for providing reaction temperature to the PCR reaction on PCR reaction chip 1 executes
Module.
Shell include upper end have opening enclosure body 11 and with enclosure body 11 be rotatablely connected and for cover close this open
The case top lid 13 of mouth, which further includes the branch for being arranged in enclosure body 11 and connecting with 11 upper end of enclosure body
Fagging 13, the installation position for installing reaction module is provided in support plate 13, and temperature control execution module is arranged under reaction module
Side.
Reaction module include for install the hinge assembly 2 of PCR reaction chip 1, the circuit board 3 being arranged on installation position and
Reaction controlling unit on circuit board 3 is set, and hinge assembly 2 is arranged above circuit board 3, and reaction controlling unit is held with temperature control
The electrical connection of row module is to control the reaction temperature that temperature control execution module provides.When case top lid 13, which is covered, closes enclosure body 11, shell
Body ontology 11 and case top lid 13 surround accommodating chamber jointly, and accommodating chamber is separated into the first accommodating chamber by support plate 13 and circuit board 3
With the second accommodating chamber, temperature control execution module is located at the second accommodating chamber, and hinge assembly 2 is located at the first accommodating chamber.
The PCR device further includes for sealing the micropore on PCR reaction chip and making mutually isolated between each micropore
Sealing element.
Shown in Figure 6, hinge assembly 2 includes that hinge on circuit board 3 and for installing PCR reaction chip is arranged in
One end rotation connection and for being pressed on sealing element on PCR reaction chip 1 the hinge lid of pedestal, one end and hinge pedestal
And the other end of hinge lid is set and is used for and the clamping of hinge pedestal or isolated hook 206.The hinge pedestal includes setting
On circuit board 3 base body 201, be arranged on base body 201 for install PCR reaction chip 1 chip installation
Block 203, the hinge lid include the hinge lid ontology 202 of one end rotation connection of one end and base body 201 and are arranged in hinge
On lid ontology 202 and for sealing element to be pressed on the chip briquetting 205 on PCR reaction chip 1,206 setting of hook is being closed
The other end of page ontology 202；Chip mounting groove 204 is opened up on chip mounting blocks 203, PCR reaction chip 1 is mounted on chip installation
In slot 204.
The temperature control execution module includes the heating list being arranged in below circuit board 3 and with 1 thermal contact conductance of PCR reaction chip
Member, the temperature being arranged in below heating unit and with the heat-sink unit of heating unit thermal contact conductance and for detecting reaction temperature pass
Sensor 7, heating unit, heat-sink unit, temperature sensor 7 are electrically connected with reaction controlling unit respectively.Reaction controlling unit is coordinated
Heat-sink unit and heating unit, which alternately cooperate, provides corresponding temperature for amplified reaction.
It is shown in Figure 5, heating unit include heat-conducting block 4, be arranged below heat-conducting block 4 and with 4 thermal contact conductance of heat-conducting block
Heater 5 and the thermally conductive block pressur plate 6 of the top of heater 5 is set, heat-conducting block 4 is between thermally conductive block pressur plate 6 and heater 5
And the upper end of heat-conducting block 4 sequentially passes through the following table face contact of thermally conductive block pressur plate 6 and circuit board 3 and PCR reaction chip 1 respectively.
The optional semiconductor heat booster of heater 5；The upper surface of heat-conducting block 4 opens up groove, and temperature sensor 7 is arranged in the groove；PCR
Carbon film is set between the contact surface of 6 the two of reaction chip 1 and heat-conducting block.
In this example, heat-conducting block 4 includes plate-like portion between thermally conductive block pressur plate 6 and heater 5 and is formed in plate-like portion
The columnar part of upper surface, columnar part sequentially pass through thermally conductive block pressur plate 6, circuit board 3 and chip mounting blocks 203 and make the upper of columnar part
The following table face contact of end and PCR reaction chip 1.Groove is provided with the upper surface of columnar part；Temperature sensor 7 is film temperature
Sensor；Carbon film is arranged between the contact surface of both the upper surface of columnar part and the lower surface of PCR reaction chip 1.
In this example, heat-sink unit include be arranged in below heater 5 and with the cooling fin 8 of 5 thermal contact conductance of heater, setting
It is below the cooling fin 8 and with the thermally conductive cover 10 of 8 thermal contact conductance of cooling fin and the fan 9 being arranged in thermally conductive cover 10.
Shown in Figure 5, power module includes being arranged in the intracorporal power panel 15 of shell, the power supply being separately positioned on shell
Line interface, power switch 14 and USB interface.Electric source line interface, power switch 14 and USB interface are arranged in enclosure body 11.
In this example, power panel 15 is located in the second accommodating chamber, and enclosure body 11 is by shell side plate 1101 and is connected to shell side plate 1101
The housing base 1102 of lower end is constituted, and shell side plate 1101 includes shell side plate body part 1101a and is arranged in shell side plate
Interface interconnecting piece 1101b on body part, electric source line interface, power switch 14 and USB interface are separately positioned on interface connecting plate
On 1101b.
The operation of the PCR device and display module include the touch display screen curtain 16 being arranged in case top lid 12 and setting
Screen control plate 17 between touch display screen curtain 16 and case top lid 12.
The PCR device further includes tweezers 18 and vacuum pencil 19, is provided in support plate 13 and is respectively used to 18 He of containing forceps
The tweezers of vacuum pencil 19 place position and vacuum pencil places position.Tweezers 18 are for clamping PCR reaction chip 1, and vacuum pencil 19 is for making
Sample and reaction solution enter in the micropore of PCR reaction chip 1, are then sealed with sealing element, make the liquid in different micropores by phase
Mutually isolation, and have one worthwhile molecule of target in each micropore, to realize the single-molecule PCR amplification of sample.
In this example, PCR reaction chip is using the microwell array containing 0.1 ten thousand~10,000,000 micropores in microwell array
Formula PCR reaction chip, the volume of each micropore are 1fL~10pL.Each micropore bottom of the PCR reaction chip is additionally provided with biochemical biography
Sensor.
Biochemical sensor can use following biochemical sensor:
As the surface of biochemical sensor is coated with hydrogen ion sensitive material, such as SiO2、Al2O3、HfO2、TiO2、Ta2O5Or (3-
Aminopropyl) triethoxysilane or phosphate anion or pyrophosphate ion sensitive material, such as phosphate ion organic film ISM
Or the molecular probes such as C4- [3] rotaxane, cyanostar.Specific such as biochemical sensor is the production of standard CMOS semiconductor technique
ISFET, that is, ion-sensitive field effect transistor.Hydrogen ion sensitive material, hydrogen ion sensitive are deposited on the metal floating gate of ISFET
Sense material can detecte out the pH value in hydrogen ion or reaction solution in PCR reaction solution.
Biochemical sensor can also using emerging nano wire as channel field effect transistor nanowire FET or
The two-dimensional semiconductors material such as person's graphene or molybdenum disulfide is the FET of channel, compares ISFET, these nanocrystal tube sensors
Higher sensitivity can be provided, so that smaller reaction cavity is provided, higher integrated level and accuracy.
Biochemical sensor can also detect hydrogen ion or phosphate anion using traditional electrode, i.e., miniature electrochemistry passes
Sensor.
The PCR device further includes the detection processing unit being arranged on circuit board 3, and detection processing unit is for acquiring biochemistry
The signal of sensor simultaneously carries out processing analysis to signal, then feeds back analysis result to operation and display module.
Hinge pedestal is equipped with the probe that electrical connection is formed with biochemical sensor pin pad, the probe and detection processing
Unit is connected to acquire the electric signal of biochemical sensor.Specifically, probe is arranged on chip mounting blocks 203.
The detection processing unit includes data acquisition circuit, ADC chip and processor, and data acquisition circuit acquires micropore bottom
The electric signal of portion's biochemical sensor after carrying out digital-to-analogue conversion by ADC chip, is conveyed to processor and carries out Digital Signal Processing and divide
Then analysis feeds back analysis result to operation and display module.Specifically, processor FPGA, DSP or ARM.
In this example, the test operation for being also used to detect the reaction solution in PCR reaction chip with display module is operated
As the result is shown.
PCR reaction not only may be implemented in the PCR device, can also realize the detection of the reaction result of PCR reaction.
The working principle of the PCR reaction of the PCR device: during using the PCR device, being given by power module should
PCR device power supply realizes the PCR reaction of sample by PCR reaction chip and temperature control execution module and reacts offer reaction to PCR
Temperature realizes setting to parameters such as the reaction time of the PCR device, temperature curve and cycle-indexes by operation and display module
It sets and shows.
The specific work process of PCR reaction: successively opening case top lid 12 and hinge lid, will be contained with pipettor and is diluted
The reaction solutions of single template samples be directly added drop-wise to the upper surface of PCR reaction chip and so that sample and reaction solution is entered PCR anti-
It answers in the micropore of chip, then the micropore and then sealing PCR conversion zone on sealing element sealing PCR reaction chip 1, close hinge
Lid, hook 206 are clamped with base body 201, and sealing element is pressed on PCR reaction chip 1 by chip briquetting 205, aobvious by touching
After the parameters such as reaction time, temperature curve and cycle-index are arranged in display screen curtain, amplified reaction can be started, in reaction process, instead
It answers control unit control heating unit and heat-sink unit to work alternatively and provides reaction temperature to reaction, be arranged when cycle-index reaches
After value, reaction is completed.
The working principle of the detection of the reaction result of the PCR reaction of the PCR device: by not carried out to reaction chip first
The pH value test of reaction solution is carried out when PCR reacts, the pH value for then carrying out reaction solution to the reaction chip of PCR after reaction is surveyed
Examination carries out data analysis according to the situation of change of secondary pH value, to obtain PCR reaction result.
For the working principle for the PCR device for better illustrating this, a specific PCR reaction chip is carried out below
The detection of PCR reaction and PCR reaction result.
PCR reaction chip is the microwell array reaction chip using CMOS ISFET as biochemical sensor, and specific targets are such as
Table 1:
The index of 1 microwell array reaction chip of table
| Sensor | ISFET |
| Micropore and number of sensors | ~26 ten thousand |
| Pore size (long, wide, depth) | 2 microns, 2 microns, 1.5 microns |
| Sensor surface inductive material type | Ta2O5 |
| Sensor surface inductive material thickness | 20 nm |
| Detect the signal type of PCR | Hydrogen ion concentration or pH variation |
Specific steps are as follows:
1, loading
The target dna solution of concentration 5pM is uniformly mixed with the PCR amplification liquid containing archaeal dna polymerase, dNTPs and primer.
Solution combined amount and ratio are as shown in table 2.Mixed liquor is added drop-wise to the microwell array PCR reaction chip with biochemical sensor
On, make to contain mixed solution in each micropore, then be sealed with sealing element, during loading, suitably dilutes solution to be measured and allow
The number of fragments of DNA in DNA solution to be measured is less than or equal to the quantity (260,000) of chip micropore, so that in micropore as far as possible
Only one DNA fragmentation.
Table 2:PCR solution is prepared
| DNA Template | 5pM |
| PCR MIX | 5ul |
| Primer | In right amount |
| Deionization H2O | To 10ul |
2, pcr amplification reaction
The chip of step 1 is placed in PCR device and carries out PCR amplification, amplification condition is as shown in table 3:
Table 3:PCR amplification condition
3. signal-obtaining
Before carrying out above-mentioned steps 2, the option program first reaction to PCR reaction is not carried out in operation and display module
Reaction solution in chip carries out pH value detection.Then to PCR reaction after reaction solution carry out pH detection, according to pH situation of change into
Then row Data Analysis Services are fed back on operating display curtain.
During carrying out data-driven to chip, each time point all generates 260,000 data, as a frame, respectively
Corresponding 260,000 biochemical sensors.(Fig. 7 is an example) as shown in Figure 7, data form one two by 512 rows × 512 column
Thermodynamic chart is tieed up, the color of each pixel corresponds to the size of the output signal of respective sensor, i.e. pH in its corresponding micropore
Value.Multiple time points can produce multiframe two dimension thermodynamic chart.
It is specific as follows:
Number frame two dimension thermodynamic chart data will be obtained in detection process, and a frame average value thermodynamic chart is acquired to the data of acquisition
And histogram Histogram analysis is carried out to it, as shown in figure 8, obtaining multiple distribution of peaks: there is no pH in the micropore of PCR
There is no variation, sensor output is 0, then corresponding micropore is negative reaction unit, i.e., negative sensor；Secondly, having single
DNA sample occurs pH in the micropore of PCR reaction and changes, then corresponding micropore is positive reaction unit, i.e., positive sensor；Also
There are some small peaks, it is to have multiple DNA samples that PCR occurs in the micropore to react that pH, which changes more greatly (sensor signal is bigger),
If solution dilution ratio is suitable, the element number of the interference is seldom.
It is 13672 that this example, which finally obtains positive number of sensors,.Illustrate there are 13672 micropores that single DNA sample has occurred
PCR reaction.
By above-mentioned setting, which is had the advantage that
1) PCR device is easy to operate simple, can be to reaction chip, sample after successively opening case top lid and hinge lid
This is operated, and is such as added drop-wise on reaction chip, installs or takes out reaction chip for the reaction solution containing sample.
2) form when PCR device injects reaction chip to sample is required without particular/special requirement, such as droplet, for sample
Injection uses general pipettor.
3) use cost of the PCR device is cheap, has microwell array chip technology based on what is be adapted to the PCR device,
Reaction solution and sample consumption needed for detection can be effectively reduced.
4) PCR device is due to being not required to configuration droplet generator, and instrument cost reduces, and compact structure, quality are lighter, side
Portable belt.
5) material of the heat-conducting block and reaction chip of the PCR device is partially hard, and when the two contacts, contact surface inevitably will appear
It is bonded not tight problem.And carbon film is partially soft relative to heat-conducting block and reaction chip hardness, and good heat conductivity, in heat-conducting block and instead
It answers chip chamber to cover certain thickness carbon film, enables to the faying face between heat-conducting block and reaction chip to be in close contact, to make
Heat transfer process it is reliable, reaction temperature is more controllable；On the other hand, also also just reduce heat without not tight problem is bonded
The loss of amount, reduces energy consumption.
6) temperature sensor is arranged in the faying face of heat-conducting block and reaction chip in the PCR device, improves reaction temperature
The accuracy of detection.
7) heat-sink unit of the PCR device includes cooling fin, fan and thermally conductive cover, can effectively improve cooling rate, thus
Shorten the reaction time.
8) PCR reaction not only may be implemented in the PCR device, can also realize the detection of PCR reaction result.
9) detection of the PCR device to PCR reaction result is used, compared to the existing detection method using fluorescence imaging,
Without take pictures, operating process is greatly simplified, and also shortens the experimental implementation time, reduces experimental cost.Simultaneously
Also overcome the not high problem of the sensitivity of existing detection method.
The above embodiments merely illustrate the technical concept and features of the present invention, and its object is to allow person skilled in the art
Scholar cans understand the content of the present invention and implement it accordingly, and it is not intended to limit the scope of the present invention.It is all according to the present invention
Equivalent change or modification made by Spirit Essence, should be covered by the protection scope of the present invention.
Claims (18)
1. a kind of portable PCR device, it is characterised in that: the PCR device includes the power module for power supply, shell, sets
Operation and display module on the housing are set, is arranged in the shell and with microwell array formula PCR reaction chip
Reaction module and the temperature for being arranged in the shell and be used to react the PCR on the PCR reaction chip offer reaction temperature
Control execution module.
2. portable PCR device according to claim 1, it is characterised in that: the reaction module includes for installing
The reaction control stated the hinge assembly of PCR reaction chip, the intracorporal circuit board of the shell is set and is arranged on the circuit board
Unit processed, the hinge assembly setting is in the circuit board, the reaction controlling unit and temperature control execution module electricity
Connection is to control the reaction temperature that the temperature control execution module provides.
3. portable PCR device according to claim 2, it is characterised in that: the PCR device further includes for sealing
It states the micropore on PCR reaction chip and makes sealing element mutually isolated between each micropore, the hinge assembly includes that setting exists
One end of on the circuit board and for installing the PCR reaction chip hinge pedestal and one end and the hinge pedestal turns
Hinge lid dynamic connecting and for being pressed on the sealing element on the PCR reaction chip.
4. portable PCR device according to claim 3, it is characterised in that: the temperature control execution module includes that setting exists
Below the circuit board and with the heating unit of the PCR reaction chip thermal contact conductance, be arranged below the heating unit and
Temperature sensor with the heat-sink unit of the heating unit thermal contact conductance and for detecting the reaction temperature, the heating are single
Member, heat-sink unit and temperature sensor are electrically connected with the reaction controlling unit respectively.
5. portable PCR device according to claim 4, it is characterised in that: the heating unit includes heat-conducting block, setting
Below the heat-conducting block and with the heater of the heat-conducting block thermal contact conductance and the heat-conducting block being arranged in above the heater
Pressing plate, the heat-conducting block is between the thermally conductive block pressur plate and heater and the upper end of the heat-conducting block sequentially passes through respectively
The thermally conductive block pressur plate and circuit board are contacted with the PCR reaction chip.
6. portable PCR device according to claim 4, it is characterised in that: the PCR reaction chip and the heating are single
Carbon film is set between both first contact surface.
7. portable PCR device according to claim 4, it is characterised in that: the upper surface of the heat-conducting block opens up groove,
The temperature sensor setting is in the groove.
8. portable PCR device according to claim 4, it is characterised in that: the heat-sink unit includes being arranged described
Below heating unit and with the cooling fin of the heating unit thermal contact conductance, be arranged in below the cooling fin and dissipated with described
The thermally conductive cover of backing thermal contact conductance and the fan being arranged in the thermally conductive cover.
9. portable PCR device according to claim 3, it is characterised in that: the PCR device further includes being arranged described
Detection processing unit on circuit board, the micropore bottom of the PCR reaction chip are provided with biochemical sensor, the detection processing
Unit is used to acquire the signal of the biochemical sensor and carries out processing analysis to signal, then feeds back analysis result to described
Operation and display module.
10. portable PCR device according to claim 9, it is characterised in that: the surface of the biochemical sensor is coated with hydrogen
Ion-sensitive material, phosphate anion or pyrophosphate ion sensitive material.
11. portable PCR device according to claim 9, it is characterised in that: the biochemical sensor is ion-sensitive field
Effect transistor, one of nano wire, graphene or crystal of molybdenum disulfide tube sensor or miniature electrochemical.
12. portable PCR device according to claim 9, it is characterised in that: the hinge pedestal is equipped with and the life
Change the probe that sensor pin pad forms electrical connection, the probe is connect with the detection processing unit to acquire the life
Change the electric signal of sensor.
13. portable PCR device according to claim 12, it is characterised in that: the detection processing unit includes data
Acquisition Circuit, ADC chip and processor, the signal of the data acquisition circuit acquisition micropore bottom biochemical sensor, by ADC core
It after piece carries out analog-to-digital conversion, is conveyed to processor and carries out Digital Signal Processing and analyze, then feed back analysis result to described
Operation and display module.
14. portable PCR device described in claim according to claim 1 ~ any one of 13, it is characterised in that: the PCR
Contain 0.1 ten thousand ~ 10,000,000 micropores in the microwell array of reaction chip, the volume of each micropore is 1fL ~ 10pL.
15. portable PCR device described in claim according to claim 1 ~ any one of 13, it is characterised in that: the shell
Body includes that upper end has the enclosure body of opening and is rotatablely connected with the enclosure body and is used to cover the shell for closing the opening
Upper cover, the operation and display module setting cover on the housing, and the PCR device further includes being arranged in the shell sheet
In vivo and the support plate that is connect with the enclosure body upper end, it is provided in the support plate for installing the reaction module
Installation position, the reaction module are arranged on the installation position, and the temperature control execution module is arranged below the reaction module.
16. portable PCR device according to claim 15, it is characterised in that: the PCR device further includes tweezers and true
Empty pen is provided in the support plate and is respectively used to place the tweezers of the tweezers and vacuum pencil and places position and vacuum pencil is placed
Position.
17. portable PCR device described in claim according to claim 1 ~ any one of 13, it is characterised in that: the electricity
Source module includes that the intracorporal power panel of the shell is arranged in, electric source line interface on the housing is respectively set and power supply is opened
It closes.
18. portable PCR device described in claim according to claim 1 ~ any one of 13, it is characterised in that: the behaviour
Make and display module includes the touch display screen curtain being arranged on the housing and is arranged in the touch display screen curtain and shell
Between screen control plate.
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