CN107807114A - Gene based on functionalization capillary is quick, quantified system analysis and method - Google Patents
Gene based on functionalization capillary is quick, quantified system analysis and method Download PDFInfo
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- CN107807114A CN107807114A CN201710815260.8A CN201710815260A CN107807114A CN 107807114 A CN107807114 A CN 107807114A CN 201710815260 A CN201710815260 A CN 201710815260A CN 107807114 A CN107807114 A CN 107807114A
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
- G01N21/6428—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes"
Abstract
The invention discloses a kind of gene based on functionalization capillary is quick, quantified system analysis and method, the system includes circulating water heating component, nucleic acid amplification reaction chamber component, optical signal excites and acquisition component and graphical analysis component, the circulating water heating component is used to be heated to assigned temperature and circulate be injected into nucleic acid amplification reaction chamber component by the water stored in storage tank, and capillary is heated and cooled down；The nucleic acid amplification reaction chamber component is used to place capillary and carry out nucleic acid amplification to it；The optical signal excites and acquisition component is used to excite the nucleic acid dye in capillary and be acquired the optical signal that it is ejected；Described image analytic unit is used to carry out quantitative analysis to the optical signal collected.The present invention has built the nucleic acid analyzer matched with capillary on the basis of Rapid nucleic acid extraction is carried out using functionalization capillary so that foranalysis of nucleic acids realizes integration, the operation of automation.
Description
Technical field
The present invention relates to technical field of biological, and in particular to a kind of gene based on functionalization capillary is quick, fixed
Measure analysis system and method.
Background technology
Gene diagnosis technology has powerful advantage in the basis of laboratory medicine and clinical research.Gene diagnosis technology has
There is the good, high sensitivity of specificity, with strong points.With making nucleic acid molecular hybridization, PCR (PCR), restricted
Restriction analysis, DNA sequencing etc. are the fast development of the gene diagnostic new technology of representative, are monitored for environmental microorganism, food security
Management, clinical disease diagnosis, criminal investigation medical jurisprudence etc. provide more abundant, effective data and information.
Wherein, PCR (PCR) technology possesses the characteristics of high sensitivity, strong adaptability, is widely used to
Gene diagnosis field.The technology is expanded using archaeal dna polymerase to the specified segment of target dna, can be by number of targets to be detected
Amount improves tens thousand of times.Using round pcr examine nucleic acid when, it is necessary first to actual sample carry out pre-treatment, with from complexity body
The higher DNA or RNA of purity is obtained in system, and removes the PCR inhibitor carried in sample, with ensure following amplification it is normal,
Effectively carry out.In addition, when detecting gene with round pcr, it is also necessary to confirm the product expanded, main method is electrophoresis at present
Method.This method to amplified production by dyeing, and using charged particle under the effect of unit electric field intensity, the unit interval moves to the interior
This principle of shifting rate difference, distinguishes target product and non-targeted product.These technical methods are not only to professional knowledge, skill set requirements
It is higher, and expensive equipment is needed, in addition, the operating process of these technical methods is extremely complex, time-consuming very long, this is just limited
Its application and development in rapid gene diagnosis.
Based on the present invention is for the functionalization capillary of genome rapid extraction, a kind of and functionalization capillary have developed
What pipe matched, for gene to be quick, instrument of quantitative analysis, realize from gene and extract amplification and subsequent detection one
Change, the operation of automation.Compared with technology compare, the step of present apparatus dramatically simplifies manual operation, not only
Human resources are saved, and avoid the experimental result mistake caused by manual operation.Meanwhile by terms of apparatus structure
Design so that overall detection time shortens dramatically, can be preferably applied for rapid gene diagnosis and analysis.
The content of the invention
The purpose of the present invention be in view of the shortcomings of the prior art with deficiency, there is provided a kind of base based on functionalization capillary
Because of quick, quantified system analysis.
Carry out that gene is quick, method of quantitative analysis another object of the present invention is to providing a kind of using said system.
The purpose of the present invention can be achieved through the following technical solutions：
A kind of gene based on functionalization capillary is quick, quantified system analysis, including circulating water heating component, nucleic acid expand
Increasing reaction chamber component, optical signal excites and acquisition component and graphical analysis component, and the circulating water heating component is used for water storage
The water of storage is heated to assigned temperature and circulated be injected into nucleic acid amplification reaction chamber component in case, capillary is heated and
Cooling；The nucleic acid amplification reaction chamber component is used to place capillary and carry out nucleic acid amplification to it；The optical signal excite and
Acquisition component is used to excite the nucleic acid dye in capillary and be acquired the optical signal that it is ejected；The figure
As analytic unit is used to carry out quantitative analysis to the optical signal collected.
Further, the circulating water heating component includes storage tank, heater, solid-state relay, temperature sensor, temperature
It is as follows to spend controller, four-way converter, water pump, water valve, fluid line and conduit coupling, specific connected mode：Three temperature
Sensor and three heaters are respectively arranged in the inside of three storage tanks, the electrical signal difference of three temperature sensors
It is connected to temperature controller channel l/2/3 signal input port, temperature controller channel l/2/3 signal output
Input of the port respectively with three solid-state relays is connected, after the output end of solid-state relay is connected with three heaters respectively
Incoming transport power supply；After the delivery port of three storage tanks is connected with water valve, four-way converter is accessed by fluid line, formed
Single stream, stream so that access nucleic acid amplification reaction chamber component water inlet, the delivery port of nucleic acid amplification reaction chamber component with
Water pump water inlet is connected, and pump outlet is connected with four-way converter, and single stream is divided into three tunnels, per the connection water of stream all the way
It is connected after valve with storage tank water inlet, forms circulating system.
Further, the storage tank is cylindrical PTFE water tanks, and diameter range is 30~60mm, altitude range 100
~180mm, moisture storage capacity are 280mL~1500mL, and top surface is provided with a blasthole, a diameter of 5~10mm, and bottom surface is provided with one and gone out
Water hole, a diameter of 5~10mm, a heater connecting hole, a diameter of 5~10mm, a temperature sensor connecting hole are a diameter of
2~6mm；The temperature controller is triple channel and the temperature controller with upper channel, can control at least three roads independent simultaneously
Heating process, using pid algorithm, export as solid-state relay pattern, input as K-type thermocouple pattern, control range is 0~
180 DEG C, control accuracy is 0.1~1 DEG C；The control signal of the solid-state relay is 3~32VDC, output signal is 24~
480VAC, electric current are 5~25A；The heater is cylindrical heater rod, and diameter range is 6~8mm, length range is 90~
170mm, operating voltage are 110~220VAC, and power is 150~220W, and bottom carries fixed thread, are connected with storage tank solid
Fixed, the diameter of thread is 5~10mm；The temperature sensor is K-type thermocouple, and diameter range is 2~5mm, length is 910~
170mm, bottom carry fixed thread, are connected with storage tank, and the diameter of thread is 2~6mm；Affiliated four-way converter diameter is
4~8mm；The water pump is peristaltic pump, and operating voltage is 12~24VDC, and power is 5~10W, maximum pump speed is 340~
800mL/min；The water valve is magnetic valve, and control voltage is 12~24VDC, and power is 0.5~1W, bear pressure for 0~
1MPa, bearing temperature are 20~100 DEG C；The fluid line is PU flexible pipes, a diameter of 4~8mm；The conduit coupling includes
SMC types thread straight-through and flexible pipe cross four-way, the screw thread scopes of SMC type thread straight-throughs are 5~10mm, connection flexible pipe a diameter of 4
~8mm, a diameter of 4~8mm of connection flexible pipe of flexible pipe cross four-way.
Further, the nucleic acid amplification reaction chamber component adds including closure, sealing ring, copper heating chamber, semiconductor
Backing, semiconductor chilling plate, temperature sensor, temperature controller, solid-state relay, the hot fin of radiating, radiator fan and part peace
Seat is filled, specific connected mode is as follows：Sealing ring is installed on the inside grooves in copper heating chamber, and closure is placed on copper heating chamber
Top, the probe of two temperature sensors is inserted laterally into the mounting hole at copper heating chamber both ends respectively, electric signal output
End is connected to temperature controller channel 1/2 signal input part, temperature controller channel 1/2 signal output
Control terminal of the port respectively with two solid-state relays is connected, the output end of solid-state relay respectively with semiconductor heating plate and half
Dc source is accessed after the series connection of conductor cooling piece；Semiconductor heating plate is placed horizontally in the upper end bottom of copper heating chamber, partly leads
Body cooling piece is placed horizontally in the lower end base of copper heating chamber, and semiconductor chilling plate lower end is placed with the hot fin of radiating, radiating heat
The air inlet of fin is placed with radiator fan, and the position that part mounting seat is used between each part is fixed.
Further, the material of the closure is PMMA, and length range is 70~100mm, width range is 40~
60mm, thickness range are 2~5mm；The sealing ring is the fluorine glue O-ring of hexagon, and line footpath be 1~3mm, internal diameter for 50~
65mm；The copper heating chamber is hexagon cavity, and the length of one pair of which relative edge is 50~70mm, in addition four edges length
Spend equal, be 15~20mm, housing depth is 3~6mm, and cavity wall thickness is 2~4mm, and cavity upper surface is along hexagon cavity
The groove of one hexagon of milling, for installing sealing ring, depth of groove is 0.5~2.5mm, and width is 1~3mm；It is described partly to lead
The length of body heating plate is 40~60mm, and width is 20~25mm, and operating voltage is 5~24VDC, and power is 28~110W；Institute
The length for stating semiconductor chilling plate is 40~60mm, and width is 20~25mm, and operating voltage is 5~24VDC, power is 58~
65W；The material of the hot fin of radiating is brass, and length is 70~100mm, and width is 15~25mm, is highly 8~15mm；Institute
The operating voltage for stating radiator fan is 5~24VDC；The material of the part mounting seat is PMMA.
Further, the optical signal excites and acquisition component includes LED excitation sources, filter group, miniature adjustable focus light
Camera is learned, specific connected mode is as follows：LED excitation sources are laterally disposed, laser is laterally projected, and filter group excites piece longitudinal
Place, and coaxial, vertical stratification state is kept with transmitting light, filter component light microscopic keeps 45 degree of angles to put with transmitting optical axis direction
Put, filter group sheet emitting is placed horizontally in the positive upper end of spectroscope, and miniature adjustable focus optical camera is positioned over filter group sheet emitting
Positive upper end.
Further, the wavelength of the LED excitation sources is 455~485nm, and power is 3~5W；The filter group includes
Piece, sheet emitting and spectroscope are excited, excites a diameter of 20~25mm of piece, thickness is 2~3mm, and wavelength is 460~490nm, hair
A diameter of 20~25mm of piece is penetrated, thickness is 2~3mm, and cutoff wavelength is 505~510nm, and spectroscope grows 30~40mm, wide by 25
~30mm, thickness are 1~2mm, and cutoff wavelength is 490~500nm；A diameter of the 5 of the miniature adjustable focus optical camera~
8mm, angle of visibility are 120 °~140 °, and the depth of field is 3~100mm, and maximum pixel is 2,000,000.
Further, it is soft to include computer, video processor, control software and image procossing for described image analytic unit
Part, the USB output port of the miniature adjustable focus optical camera are connected with computer, and the video processor is optical camera
Head adaptation processor, the control software and image processing software are Labview softwares.
Another object of the present invention can be achieved through the following technical solutions：
It is a kind of to carry out that gene is quick, method of quantitative analysis using said system, it the described method comprises the following steps：
(1) after separation and extraction that nucleic acid, is carried out using functionalization capillary, fluorescent quantitation is passed through into capillary
PCR reaction mixtures, and capillary is sealed；
(2) capillary of good seal, is put into nucleic acid amplification reaction intracavitary, nucleic acid amplification reaction chamber is sealed, forms envelope
The liquid flow path closed；
(3), by the denaturation required for computer operation interface setting nucleic acid amplification, annealing, elongating temperature with it is corresponding
Duration and period；
(4) after, the water temperature that system is automatically detected in each storage tank reaches design temperature, matching somebody with somebody for water pump and water valve is passed through
Closing operation, the water of different temperatures is flowed successively through nucleic acid amplification reaction chamber, reach amplification purpose；
(5), in circulating each time, after the current in elongating temperature are by nucleic acid amplification reaction 20~25s of chamber, it is
System opens LED excitation sources and miniature adjustable focus optical camera, and IMAQ is carried out to the optical signalling in each capillary；
(6), miniature adjustable focus optical camera transmits the optical signalling collected to computer, and system automatically analyzes respectively
Optical signal intensity in individual capillary；
(7) after the completion of, expanding, automatically by different cycle-indexes, the optical signal intensity of different capillaries is carried out system
Totally collect, export quantitative amplification experimental result；
(8), the semiconductor heating plate and semiconductor chilling plate cooperation of system control nucleic acid amplification reaction chamber bottom, makes
Reaction chamber is internally formed thermograde；
(9) after, thermograde is stable, LED excitation sources and miniature adjustable focus optical camera are opened, to each capillary
Interior optical signalling carries out IMAQ；
(10), miniature adjustable focus optical camera, which transmits the optical signalling collected to computer, system, automatically analyzes
Optical signal intensity in each capillary；
(11) optical signal intensity of diverse location in each capillary, is measured, and data are collected, output is molten
Solution curve analysis result.
The present invention compared with prior art, has the following advantages that and beneficial effect：
1st, the present invention has been built and capillary on the basis that Rapid nucleic acid extraction is carried out using functionalization capillary
The nucleic acid analyzer matched somebody with somebody so that foranalysis of nucleic acids realizes integration, the operation of automation, and artificial behaviour has been reduced to a great extent
Make, avoid the error in the experimental result caused by human factor.
2nd, the present invention largely shortens core by carrying out the design in structure to nucleic acid amplification thermal response chamber component
It is the acid detection required time, compact-sized, easy to operate beneficial to the scene of carrying out quickly quantification detection, it is easy to promote and answers
With.
Brief description of the drawings
Fig. 1 is that a kind of gene based on functionalization capillary is quick in the embodiment of the present invention 1, the structure of quantified system analysis
Schematic diagram.
Fig. 2 is the structural representation of the amplifying nucleic acid amplified reaction chamber component of the embodiment of the present invention 1.
Fig. 3 is excited for optical signal in the embodiment of the present invention 1 and the structural representation of acquisition component.
Fig. 4 (a) is the analysis result figure of the quantitative amplification curve that system acquisition arrives in the embodiment of the present invention 2, and Fig. 4 (b) is
The analysis result figure for the solubility curve that system acquisition arrives in embodiment 2.
Embodiment
With reference to embodiment and accompanying drawing, the present invention is described in further detail, but embodiments of the present invention are unlimited
In this.
Embodiment 1：
Present embodiments provide that a kind of gene based on functionalization capillary is quick, quantified system analysis, structural representation
As shown in figure 1, being excited including circulating water heating component, nucleic acid amplification reaction chamber component, optical signal and acquisition component and image point
Component is analysed, the circulating water heating component is used to the water stored in storage tank is heated to assigned temperature and circulated to be injected into nucleic acid
In amplified reaction chamber component, capillary is heated and cooled down；The nucleic acid amplification reaction chamber component is used to place capillary
And nucleic acid amplification is carried out to it；The optical signal excites and acquisition component is used to excite simultaneously the nucleic acid dye in capillary
The optical signal that it is ejected is acquired；Described image analytic unit is used to the optical signal collected is carried out quantifying to divide
Analysis.
Wherein, the circulating water heating component includes storage tank, heater, solid-state relay, temperature sensor, temperature control
Device, four-way converter, water pump, water valve, fluid line and conduit coupling processed, specific connected mode are as follows：Three TEMPs
Device and three heaters are respectively arranged in the inside of three storage tanks, and the electrical signal of three temperature sensors connects respectively
In temperature controller channel l/2/3 signal input port, temperature controller channel l/2/3 signal output port
The input with three solid-state relays is connected respectively, and the output end of solid-state relay accesses after being connected respectively with three heaters
AC power；After the delivery port of three storage tanks is connected with water valve, four-way converter is accessed by fluid line, formed single
Stream, stream and then the water inlet for accessing nucleic acid amplification reaction chamber component, the delivery port and water pump of nucleic acid amplification reaction chamber component
Water inlet is connected, and pump outlet is connected with four-way converter, and single stream is divided into three tunnels, after stream connecting with water valve all the way
It is connected with storage tank water inlet, forms circulating system.
The structural representation of the nucleic acid amplification reaction chamber component as shown in Fig. 2 including closure, sealing ring, it is copper plus
Hot chamber, semiconductor heating plate, semiconductor chilling plate, temperature sensor, temperature controller, solid-state relay, the hot fin of radiating, radiating
Fan and part mounting seat, specific connected mode are as follows：Sealing ring is installed on the inside grooves in copper heating chamber, closure lid
In the top of copper heating chamber, the probe of two temperature sensors is inserted laterally into the mounting hole at copper heating chamber both ends respectively
Interior, electrical signal is connected to temperature controller channel 1/2 signal input part, temperature controller channel
Control terminal of 1/2 signal output port respectively with two solid-state relays is connected, and the output end of solid-state relay is respectively with half
Dc source is accessed after conductor heating plate and semiconductor chilling plate series connection；Semiconductor heating plate is placed horizontally in copper heating chamber
Upper end bottom, semiconductor chilling plate are placed horizontally in the lower end base of copper heating chamber, and semiconductor chilling plate lower end is placed with scattered
Hot fin, the air inlet for the hot fin that radiates are placed with radiator fan, and the position that part mounting seat is used between each part is fixed.
The optical signal excite and the structural representation of acquisition component as shown in figure 3, including LED excitation sources, filter group,
Miniature adjustable focus optical camera, specific connected mode are as follows：LED excitation sources are laterally disposed, laser is laterally projected, filter
Group excites piece placed longitudinally, and keeps coaxial, vertical stratification state with transmitting light, and filter component light microscopic is kept with transmitting optical axis direction
45 degree of angles are placed, and filter group sheet emitting is placed horizontally in the positive upper end of spectroscope, and miniature adjustable focus optical camera is positioned over filter
The positive upper end of microscope group sheet emitting.
Described image analytic unit includes computer, video processor, control software and image processing software, described miniature
The USB output port of adjustable focus optical camera is connected with computer, and the video processor is optical camera adaptation processing
Device, the control software and image processing software are Labview softwares.
Embodiment 2：
Present embodiments provide it is a kind of salmonella is carried out using system described in embodiment 1 gene quickly, quantitative analysis
Method, the reagent used in methods described is purchased from New England biotech companies, comprises the following steps：
(1) after the separation and extraction of nucleic acid that salmonella, is carried out using functionalization capillary, it is passed through into capillary
Quantitative fluorescent PCR reaction mixture, including Taq enzyme, primer, dNTP the and PCR buffer of corresponding salmonella, and
Capillary is sealed；
(2) capillary of good seal, is put into nucleic acid amplification reaction intracavitary, nucleic acid amplification reaction chamber is sealed, forms envelope
The liquid flow path closed；
(3), by the denaturation required for computer operation interface setting nucleic acid amplification, annealing, elongating temperature with it is corresponding
Duration and period, set herein unwind, anneal, elongating temperature is 95 DEG C, 54 DEG C, 72 DEG C respectively, the duration is equal
For 30s, period is 40 circulations；
(4) after, the water temperature that system is automatically detected in each storage tank reaches design temperature, matching somebody with somebody for water pump and water valve is passed through
Closing operation, the water of different temperatures is flowed successively through nucleic acid amplification reaction chamber, reach amplification purpose；
(5), in circulating each time, after the current in elongating temperature are by nucleic acid amplification reaction 20~25s of chamber, it is
System opens LED excitation sources and miniature adjustable focus optical camera, and IMAQ is carried out to the optical signalling in each capillary；
(6), miniature adjustable focus optical camera transmits the optical signalling collected to computer, and system automatically analyzes respectively
Optical signal intensity in individual capillary；
(7) after the completion of, expanding, automatically by different cycle-indexes, the optical signal intensity of different capillaries is carried out system
Totally collect, quantitative amplification experimental result is exported, shown in analysis of experimental results figure such as Fig. 4 (a)；
(8), the semiconductor heating plate and semiconductor chilling plate cooperation of system control nucleic acid amplification reaction chamber bottom, makes
Reaction chamber is internally formed thermograde, and what is be here formed as is 75 to 95 DEG C of thermograde；
(9) after, thermograde is stable, LED excitation sources and miniature adjustable focus optical camera are opened, to each capillary
Interior optical signalling carries out IMAQ；
(10), miniature adjustable focus optical camera, which transmits the optical signalling collected to computer, system, automatically analyzes
Optical signal intensity in each capillary；
(11) optical signal intensity of diverse location in each capillary, is measured, and data are collected, output is molten
Solution curve analysis result, as shown in Fig. 4 (b).
It is described above, patent preferred embodiment only of the present invention, but the protection domain of patent of the present invention is not limited to
This, any one skilled in the art is in the scope disclosed in patent of the present invention, according to the skill of patent of the present invention
Art scheme and its patent of invention design are subject to equivalent substitution or change, belong to the protection domain of patent of the present invention.
Claims (9)
1. a kind of gene based on functionalization capillary is quick, quantified system analysis, it is characterised in that：The system includes circulation
Water heating component, nucleic acid amplification reaction chamber component, optical signal excite and acquisition component and graphical analysis component, the recirculated water add
Hot component is used to be heated to assigned temperature and circulate be injected into nucleic acid amplification reaction chamber component by the water stored in storage tank, right
Capillary is heated and cooled down；The nucleic acid amplification reaction chamber component is used to place capillary and carry out nucleic acid amplification to it；
The light that the optical signal excites and acquisition component is used to excite the nucleic acid dye in capillary and eject it is believed
Number it is acquired；Described image analytic unit is used to carry out quantitative analysis to the optical signal collected.
2. a kind of gene based on functionalization capillary according to claim 1 is quick, quantified system analysis, its feature exists
In：The circulating water heating component includes storage tank, heater, solid-state relay, temperature sensor, temperature controller, four-way
Converter, water pump, water valve, fluid line and conduit coupling, specific connected mode are as follows：Three temperature sensors and three plus
Hot device is respectively arranged in the inside of three storage tanks, and the electrical signal of three temperature sensors is connected to temperature control
Device channel l/2/3 signal input port, temperature controller channel l/2/3 signal output port respectively with three
The input connection of solid-state relay, the output end of solid-state relay access AC power after being connected respectively with three heaters；
After the delivery port of three storage tanks is connected with water valve, four-way converter is accessed by fluid line, forms single stream, stream
And then the water inlet of nucleic acid amplification reaction chamber component is accessed, delivery port and the water pump water inlet of nucleic acid amplification reaction chamber component connect
Connect, pump outlet is connected with four-way converter, and single stream is divided into three tunnels, per stream connecting with water valve all the way after with storage tank
Water inlet connects, and forms circulating system.
3. a kind of gene based on functionalization capillary according to claim 2 is quick, quantified system analysis, its feature exists
In：The storage tank is cylindrical PTFE water tanks, and diameter range is 30~60mm, and altitude range is 100~180mm, moisture storage capacity
For 280mL~1500mL, top surface is provided with a blasthole, a diameter of 5~10mm, and bottom surface is provided with an apopore, a diameter of 5~
10mm, a heater connecting hole, a diameter of 5~10mm, a temperature sensor connecting hole, a diameter of 2~6mm；The temperature
It is triple channel and the temperature controller with upper channel to spend controller, can control the heating process of at least three tunnel independences simultaneously, adopt
With pid algorithm, export as solid-state relay pattern, it is 0~180 DEG C to input as K-type thermocouple pattern, control range, control essence
Spend for 0.1~1 DEG C；The control signal of the solid-state relay is 3~32VDC, and output signal is 24~480VAC, electric current 5
~25A；The heater is cylindrical heater rod, and diameter range is 6~8mm, and length range is 90~170mm, operating voltage
For 110~220VAC, power is 150~220W, and bottom carries fixed thread, is connected with storage tank, the diameter of thread be 5~
10mm；The temperature sensor is K-type thermocouple, and diameter range is 2~5mm, and length is 910~170mm, and bottom is with fixation
Screw thread, it is connected with storage tank, the diameter of thread is 2~6mm；Affiliated four-way converter diameter is 4~8mm；The water pump is
Peristaltic pump, operating voltage are 12~24VDC, and power is 5~10W, and maximum pump speed is 340~800mL/min；The water valve is electricity
Magnet valve, control voltage are 12~24VDC, and power is 0.5~1W, and it is 0~1MPa to bear pressure, and bearing temperature is 20~100 DEG C；
The fluid line is PU flexible pipes, a diameter of 4~8mm；The conduit coupling includes SMC types thread straight-through and flexible pipe cross four
Logical, the screw thread scopes of SMC type thread straight-throughs is 5~10mm, a diameter of 4~8mm of connection flexible pipe, and the connection of flexible pipe cross four-way is soft
Pipe diameter is 4~8mm.
4. a kind of gene based on functionalization capillary according to claim 1 is quick, quantified system analysis, its feature exists
In：The nucleic acid amplification reaction chamber component includes closure, sealing ring, copper heating chamber, semiconductor heating plate, semiconductor refrigerating
Piece, temperature sensor, temperature controller, solid-state relay, radiating hot fin, radiator fan and part mounting seat, specific connection side
Formula is as follows：Sealing ring is installed on the inside grooves in copper heating chamber, and closure is placed on the top of copper heating chamber, two temperature
The probe of sensor is inserted laterally into the mounting hole at copper heating chamber both ends respectively, and electrical signal is connected to temperature
Controller channel 1/2 signal input part, temperature controller channel 1/2 signal output port are solid with two respectively
The control terminal connection of state relay, after the output end of solid-state relay is connected with semiconductor heating plate and semiconductor chilling plate respectively
Access dc source；Semiconductor heating plate is placed horizontally in the upper end bottom of copper heating chamber, and semiconductor chilling plate is laterally disposed
In the lower end base of copper heating chamber, semiconductor chilling plate lower end is placed with the hot fin of radiating, and the air inlet for the hot fin that radiates is placed with
Radiator fan, the position that part mounting seat is used between each part are fixed.
5. a kind of gene based on functionalization capillary according to claim 4 is quick, quantified system analysis, its feature exists
In：The material of the closure is PMMA, and length range is 70~100mm, and width range is 40~60mm, thickness range 2
~5mm；The sealing ring is the fluorine glue O-ring of hexagon, and line footpath is 1~3mm, and internal diameter is 50~65mm；The copper heating
Chamber is hexagon cavity, and the length of one pair of which relative edge is 50~70mm, in addition four edges equal length, be 15~
20mm, housing depth are 3~6mm, and cavity wall thickness is 2~4mm, and cavity upper surface is along one hexagon of hexagon cavity milling
Groove, for installing sealing ring, depth of groove is 0.5~2.5mm, and width is 1~3mm；The length of the semiconductor heating plate
For 40~60mm, width is 20~25mm, and operating voltage is 5~24VDC, and power is 28~110W；The semiconductor chilling plate
Length be 40~60mm, width is 20~25mm, and operating voltage is 5~24VDC, and power is 58~65W；The hot fin of radiating
Material be brass, length is 70~100mm, and width is 15~25mm, is highly 8~15mm；The work of the radiator fan
Voltage is 5~24VDC；The material of the part mounting seat is PMMA.
6. a kind of gene based on functionalization capillary according to claim 1 is quick, quantified system analysis, its feature exists
In：The optical signal excites and acquisition component includes LED excitation sources, filter group, miniature adjustable focus optical camera, specifically connects
It is as follows to connect mode：LED excitation sources are laterally disposed, laser is laterally projected, and filter group excites piece placed longitudinally, and with launching light
Coaxial, vertical stratification state is kept, filter component light microscopic keeps 45 degree of angles to place with transmitting optical axis direction, and filter group sheet emitting is horizontal
To the positive upper end of spectroscope is positioned over, miniature adjustable focus optical camera is positioned over the positive upper end of filter group sheet emitting.
7. a kind of gene based on functionalization capillary according to claim 6 is quick, quantified system analysis, its feature exists
In：The wavelength of the LED excitation sources is 455~485nm, and power is 3~5W；The filter group includes exciting piece, sheet emitting
And spectroscope, excite a diameter of 20~25mm of piece, thickness is 2~3mm, and wavelength is 460~490nm, sheet emitting it is a diameter of
20~25mm, thickness are 2~3mm, and cutoff wavelength is 505~510nm, and spectroscope grows 30~40mm, wide 25~30mm, and thickness is
1~2mm, cutoff wavelength are 490~500nm；A diameter of 5~8mm of the miniature adjustable focus optical camera, angle of visibility are
120 °~140 °, the depth of field is 3~100mm, and maximum pixel is 2,000,000.
8. a kind of gene based on functionalization capillary according to claim 1 is quick, quantified system analysis, its feature exists
In：Described image analytic unit includes computer, video processor, control software and image processing software, described miniature adjustable
The USB output port of burnt optical camera is connected with computer, and the video processor is optical camera adaptation processor, institute
It is Labview softwares to state control software and image processing software.
Carry out that gene is quick, method of quantitative analysis using system described in claim 1 9. a kind of, it is characterised in that the side
Method comprises the following steps：
(1) after separation and extraction that nucleic acid, is carried out using functionalization capillary, it is anti-that quantitative fluorescent PCR is passed through into capillary
Mixed liquor is answered, and capillary is sealed；
(2) capillary of good seal, is put into nucleic acid amplification reaction intracavitary, nucleic acid amplification reaction chamber is sealed, forms closing
Liquid flow path；
(3), continued by the denaturation required for computer operation interface setting nucleic acid amplification, annealing, elongating temperature with corresponding
Time and period；
(4) after, the water temperature that system is automatically detected in each storage tank reaches design temperature, grasped by the cooperation of water pump and water valve
Make, the water of different temperatures is flowed successively through nucleic acid amplification reaction chamber, reach amplification purpose；
(5), in circulating each time, after the current in elongating temperature are by nucleic acid amplification reaction 20~25s of chamber, system is beaten
LED excitation sources and miniature adjustable focus optical camera are opened, IMAQ is carried out to the optical signalling in each capillary；
(6), miniature adjustable focus optical camera transmits the optical signalling collected to computer, and system automatically analyzes each hair
Optical signal intensity in tubule；
(7) after the completion of, expanding, automatically by different cycle-indexes, the optical signal intensity of different capillaries is carried out totally system
Collect, export quantitative amplification experimental result；
(8), the semiconductor heating plate and semiconductor chilling plate cooperation of system control nucleic acid amplification reaction chamber bottom, makes reaction
Chamber is internally formed thermograde；
(9) after, thermograde is stable, LED excitation sources and miniature adjustable focus optical camera are opened, in each capillary
Optical signalling carries out IMAQ；
(10), miniature adjustable focus optical camera transmits the optical signalling collected automatically analyzes each to computer, system
Optical signal intensity in capillary；
(11) optical signal intensity of diverse location in each capillary, is measured, and data are collected, output dissolving is bent
Line analysis result.
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Application publication date: 20180316 |