CN104630373B - A kind of fast parallel detection method of micro-fluidic chip nucleic acid and system - Google Patents
A kind of fast parallel detection method of micro-fluidic chip nucleic acid and system Download PDFInfo
- Publication number
- CN104630373B CN104630373B CN201510078655.5A CN201510078655A CN104630373B CN 104630373 B CN104630373 B CN 104630373B CN 201510078655 A CN201510078655 A CN 201510078655A CN 104630373 B CN104630373 B CN 104630373B
- Authority
- CN
- China
- Prior art keywords
- micro
- fluidic chip
- nucleic acid
- detection
- reaction
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Classifications
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
- B01J19/00—Chemical, physical or physico-chemical processes in general; Their relevant apparatus
- B01J19/0046—Sequential or parallel reactions, e.g. for the synthesis of polypeptides or polynucleotides; Apparatus and devices for combinatorial chemistry or for making molecular arrays
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6813—Hybridisation assays
- C12Q1/6834—Enzymatic or biochemical coupling of nucleic acids to a solid phase
- C12Q1/6837—Enzymatic or biochemical coupling of nucleic acids to a solid phase using probe arrays or probe chips
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Engineering & Computer Science (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Analytical Chemistry (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Biotechnology (AREA)
- Immunology (AREA)
- Microbiology (AREA)
- Molecular Biology (AREA)
- Physics & Mathematics (AREA)
- Biophysics (AREA)
- Biochemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Investigating, Analyzing Materials By Fluorescence Or Luminescence (AREA)
- Investigating Or Analysing Materials By The Use Of Chemical Reactions (AREA)
Abstract
The present invention relates to a kind of fast parallel detection method of micro-fluidic chip nucleic acid and system, including herein below：The one micro-fluidic chip nucleic acid detection system for including micro-fluidic chip, motor, exciting light, bifocal segment imaging lens group, detector, signal acquisition process device and display is set；Micro-fluidic chip includes more than one reaction channel, and heating film is provided with around micro-fluidic chip, and the air layer of submillimeter is kept between micro-fluidic chip and heating film；Using exciting light irradiation micro-fluidic chip, make nucleic acid samples generation fluorescence under the exciting of exciting light, analog signal is produced on the detector through bifocal segment imaging lens group convergence fluorescence, caused analog signal is sent to signal acquisition process device generation real-time fluorescence detection signal by detector, and shows real-time fluorescence detection signal by display.The present invention can be used for the fields such as the diagnosis of analysis of clinic pathogenic microorganism molecule, Food Inspection quarantine, food poisoning detection of pathogens, bacteriology classification and epidemiology survey, have larger economic and social benefits.
Description
Technical field
It is fast parallel especially with regard to a kind of micro-fluidic chip nucleic acid the present invention relates to a kind of nucleic acid detection method and system
Detection method and system.
Background technology
Micro-full analytical system (μ TAS：Micro-separation in Miniaturized Total Analysis
System it is) the last advanced bio-medical analysis technology risen of twentieth century, it detects sample preparation, biochemical reaction and result
Deng process integration into one piece of slight micro-fluidic chip, realize from sample collection to result report it is integrated, automation,
Miniaturization.
Sample preparation is first link of micro-full analytical system, because the species of primary sample is various, such as blood, phlegm
Liquid, saliva, oral cavity test paper, urine, excrement, biological tissue cell, pathogen and its organism of infection etc., thus cause sample
Preparation implements relatively complicated, lacks unified standard, still passes through discrete cascade using a variety of legacy equipments at present
Mode realize.
Biochemical reaction mainly has alternating temperature amplification and two big type of isothermal duplication.Alternating temperature amplification method (Polymerase
Chain reaction, PCR) it is earliest nucleic acid amplification technologies, startup nucleic acid is matched by positive and negative two special primers completely
Amplified reaction, circulated in units of expanding the cycle, each about 90 seconds cycle of amplification, including denaturation stage, annealing
Stage and extension stage, wherein, denaturation stage is denatured 15 seconds under 94 DEG C of high temperature, and annealing stage is moved back under 60 DEG C of low temperature
Fire 30 seconds, extension stage are to synthesize starting point as nucleic acid using primer by the effect of enzyme at 72 DEG C along template direction to extend 45 seconds
Clock.Amplification procedure is exactly that cycle constantly circulation is expanded as tens or more to complete.In alternating temperature amplification method, one
The individual amplification cycle only just carries out nucleic acid synthesis amplification in the extension stage, and denaturation stage and annealing stage simply synthesize for nucleic acid
Amplification is prepared, and therefore, is calculated from the time, and alternating temperature amplification method only accounts for full-time for the effective time of nucleic acid amplification
50%.In order to improve nucleic acid amplification efficiency and improve temperature-controlled conditions, people successively have studied a variety of isothermal amplifications again,
As Walker GT et al. 1992 report strand displacement amplification (Strand displacement amplification,
SDA) method, Liu D et al. are in rolling circle amplification (Rolling circle amplification, the RCA) side of report in 1996
Ring mediated isothermal amplification (the Loop-mediated isothermal of method and Tsugunori Notomi et al. in report in 2000
Amplification, LAMP) method etc..It is different from alternating temperature amplification method, isothermal amplification method whole process be maintained at one it is fixed
At a temperature of, without high-temperature denatured and process annealing process, in the absence of time loss caused by temperature change, therefore its amplification rate
It is very fast, can be in the short period of time by target nucleic acid amplification to 109-1010Individual copy.And calculated from the time, isothermal expands
Increasing method All Time is carrying out nucleic acid amplification, and time availability reaches 100%, so isothermal amplification method is with higher
Nucleic acid amplification efficiency.
As a result detecting mainly has fluorescent method, turbidity method etc., the usual reduced turbidity method of detection sensitivity of fluorescent method
Height, its system are also more relative complex.In fluorescent method, real-time fluorescence quantitative PCR instrument is exactly that wherein important nucleic acid amplification is real
When detecting instrument, but at present this quasi-instrument mainly using Tube manage, it is necessary to which 20 μ L above reaction systems, detection sensitivity are usual
More than 1000 nucleic acid molecules copy numbers, also costly, an instrument needs 20~300,000 yuan to instrument price.The country has more
Family's unit surrounds the research work for having carried out detection method based on the detection of test tube real-time fluorescence quantitative PCR, such as the grand science and technology in Xi'an day
Co., Ltd (patent No. ZL200410073432.1), Da'an Gene Company, Zhongshan University's (patent No.
ZL200420102062.5) etc., their inventive point is in the pcr amplification reaction in using different fiber-optic coupling methods to test tube
Product carries out real time fluorescent quantitative detection, reaches simplified light path system structure, reduces the purpose of whole instrument cost.But
Because the coupling for being related to optical fiber is transmitted with signal, the numerical aperture of fluorescence signal receiving terminal can not possibly be very high, and fluorescence signal exists
With the increase of distance, its loss significantly increases when being transmitted in optical fiber, therefore the detection sensitivity of instrument is limited by larger.
In order to improve detection sensitivity and reduce the consumption of sample and reagent, a kind of micro-nano system has been invented by Boao Biological Co., Ltd
Fluid chip detecting system, using 12 lens sets focal plane imaging Systems for optical inspection structure in pairs, realize micro-fluidic chip
Nucleic acid isothermal amplification is analyzed, and detection sample consumption reaches below 10 μ L.But, the system is relatively complicated, equipment instrument ratio
Larger, sample introduction still needs to use external independent centrifuge, and due to being open fan cooling, it is desirable to environment temperature is low
In 30 DEG C, humidity is not higher than 80%, is mainly suitable for using in laboratory environment.In order to meet family, community medical therapy unit, family
Outside, marine and the special environment condition such as earthquake or typhoon disaster area requirement, is highly desirable to develop a kind of miniature portable
Formula, looser to temperature, salt fog and humidity requirement, operation is simpler, also can in the outlying mountain area that net electricity interrupts or net electricity is obstructed
The fast parallel detecting instrument of Portable disease opportunistic pathogen used.
The content of the invention
In view of the above-mentioned problems, it is an object of the invention to provide one kind trace sample can be realized it is quick, parallel, accurate,
The fast parallel detection method of micro-fluidic chip nucleic acid and system of efficient pathogen foranalysis of nucleic acids identification.
To achieve the above object, the present invention takes following technical scheme：A kind of fast parallel detection of micro-fluidic chip nucleic acid
Method, comprise the following steps：1) one is set to include micro-fluidic chip, motor, exciting light, bifocal segment imaging lens group, detection
Device, signal acquisition process device, temperature control modules, the micro-fluidic chip nucleic acid detection system of motion-control module and display；
Wherein, micro-fluidic chip includes more than one reaction channel, heating film is provided with around micro-fluidic chip, micro-fluidic chip is with adding
The air layer of submillimeter is kept between hotting mask；2) according to some indicator nucleic acids to be detected, by corresponding detection of nucleic acids molecule
Probe is separately fixed at the reaction channel bottom of micro-fluidic chip；3) nucleic acid samples being analysed to dissolve in detection of nucleic acids reagent
In, the detection of nucleic acids reagent for having dissolved in nucleic acid samples is injected accordingly from each sample holes of micro-fluidic chip using pipettor
Microfluidic channel；4) signal acquisition process device realizes that high speed rotating centrifugal revolves with low speed by moving control module for controlling motor
Turn to scan and deposit, signal acquisition process device drives micro-fluidic chip to rotate at a high speed in the sample introduction stage by controlled motor, makes miniflow
Nucleic acid samples in control passage are respectively enterd in the reaction channel of micro-fluidic chip automatically by high speed rotating centrifugal；Complete sample introduction
Afterwards, signal acquisition process device is scanned by the same motor driven micro-fluidic chip low speed rotation of moving control module for controlling, is realized
To the fast parallel detection of multiple reaction channels on micro-fluidic chip；Synchronous signal Acquisition Processor passes through temperature control modules control
Heating film processed, the bath flowing heating of submillimeter laminar air is carried out to micro-fluidic chip, micro-fluidic chip is simultaneously under motor driving
Rotate, drive air flow so that the temperature on micro-fluidic chip between reaction channel is uniform；Led to according to practical application
Excess temperature control module control micro-fluidic chip is maintained at a certain temperature in 40 DEG C~90 DEG C temperature ranges so that micro-fluidic core
The primer of each reaction channel is released in piece, and is mixed with nucleic acid samples and detection of nucleic acids with reagent, in isothermal duplication condition
Lower carry out nucleic acid amplification reaction, realize that trace nucleic acid samples molecule diagnoses；5) using light irradiation micro-fluidic chip is excited, miniflow is made
The generation fluorescence under the exciting of exciting light of the nucleic acid samples in chip is controlled, phosphor collection efficiency is made by bifocal segment imaging lens group
Reach optical diffraction limit, the fluorescence being emitted through bifocal segment imaging lens group is accumulated produces analog signal on the detector, visits
Survey device and caused analog signal is sent to signal acquisition process device generation real-time fluorescence detection signal, and shown by display
Real-time fluorescence detection signal.
The reaction that step 2) the amplifying nucleic acid detection is fixed on micro-fluidic chip with molecular probe by adsorption is led to
Road bottom, or fixed using the Bc material embedding of agarose or oligosaccharide or fusing point in the range of 40 DEG C~90 DEG C
In the reaction channel bottom of micro-fluidic chip.
The detection of nucleic acids is made up of with molecular probe 6 sections of primers, is detected for different indicator nucleic acids, and 6 sections of primers are set
Count into the nucleic acid fragment of different A, T, G, C base sequence orders and length；Detection of nucleic acids with reagent mainly by dNTPs,
25 DEG C of EvaGreen, DTT, BstDNA Polymerase Buffer, pH 8.8at Tris-HCl, MgSO4, M-MLV
Reverse transcriptase, RNasin Plus, BstDNA Polymerase and Betaine are formed, and overall reaction system≤
1.5μL。
The step 3) will dissolve in the detection of nucleic acids reagents of nucleic acid samples from micro-fluidic chip using pipettor
The detailed process that each sample holes inject corresponding microfluidic channel is：Each sample holes and pipettor gun head on micro-fluidic chip are tight
Close fit, preset one section of air column is on the top of pipettor during sample introduction.
Rotating speed >=6000 rev/min of the micro-fluidic chip under motor driving during high speed rotating centrifugal sample introduction in the step 4)
Clock；Micro-fluidic chip progress low speed rotation scanning under the driving of same motor, rotating speed≤2 rev/min of low speed rotation scanning,
Precise angular positions deviation≤0.1 °.
A kind of fast parallel detecting system of micro-fluidic chip nucleic acid, it is characterised in that：It includes micro-fluidic chip, Spring Card
Piece location structure, pulley drive structure, motor, motion-control module, heating film, temperature control module, display, power supply, signal are adopted
Set processor, semiconductor refrigerating light source module, condenser, exciter filter, dichroscope, speculum, object lens, imaging len,
Launch optical filter, pinhole diaphragm and detector；The micro-fluidic chip is fixed by the spring card location structure, institute
State spring card location structure and the motor, motion control mould described in the motor connection are connected by the pulley drive structure
Block；The heating film is set around the micro-fluidic chip, submillimeter is kept between the micro-fluidic chip and the heating film
Air layer, the heating film connects the temperature control module；The motion-control module, temperature control module and display are connected to
The signal acquisition process device；The signal acquisition process device sends motion control signal to the motion-control module, institute
State motion-control module and the motor is controlled according to the motion control signal received, the motor passes through the belt wheel transmission knot
Structure drives the micro-fluidic chip to rotate；Meanwhile temperature control signals are delivered to the temperature control by the signal acquisition process device
Module, the temperature control module control the heating film according to the temperature control signals received, reach the micro-fluidic chip
The temperature that biological sample reaction requires；The power supply is used to be powered with single device to be each, the power supply ES using net electricity,
The power supply mode of solar energy and battery compatibility；The signal acquisition process device controls the semiconductor refrigerating light source module transmitting to swash
Luminous, the exciting light of transmitting is transmitted into after condenser collimation by the semiconductor refrigerating light source module described excites optical filtering
Piece, exciting light is filtered into after quasi- monochromatic excitation light and is transmitted into speculum through the dichroscope by the exciter filter, through institute
The exciting light for stating speculum reflection irradiates the reaction channel of the micro-fluidic chip by object lens convergence, excites the miniflow
Control biological sample in chip reaction channel and produce fluorescence；The fluorescence is transmitted into institute after object lens convergence is collimated into directional light
Speculum is stated, the fluorescent emission through speculum reflection to the dichroscope, the fluorescence through dichroscope reflection is sent out
The transmitting optical filter is mapped to filter off except after the exciting light in fluorescence, the pinhole diaphragm is converged to through the imaging len,
Fluorescence after the pinhole diaphragm removes veiling glare is received by the detector, and the detector will detect the signal hair received
The signal acquisition process device generation real-time fluorescence detection signal is sent to, and real-time fluorescence detection letter is shown by the display
Number.
The micro-fluidic chip includes a bottom plate, the centrally disposed positioning round orifice of bottom plate, the side of the positioning round orifice
Edge sets a detent；It is provided with the microfluidic channel of one or more on the bottom plate around the positioning round orifice, every
The microfluidic channel includes a microchannel, and some reaction channels are provided with each microchannel, each described micro- logical
The one end in road sets a sample holes, and the other end of each microchannel sets a venthole, micro- before each venthole
Buffer pool is provided with passage；Wherein, the reaction channel also compares comprising positive quality control control reaction passage and negative Quality Control
Reaction channel, the positive quality control control reaction passage are fixed with the standard quality-control product index nucleic acid spy that can produce fluorescence signal
Pin, the negative Quality Control control reaction passage are fixed with the standard quality-control product index probe either sky that can not produce fluorescence signal
White probe.
The spring card location structure includes a positioning seat, a locating dowel and the forced snap lock mandrel of three hemisphericals；
The through hole that the positioning seat centrally disposed one is attached with the belt wheel mechanism, the positioning seat side fixed setting and institute
The locating dowel that detent is used cooperatively is stated, divides equally around the positioning seat in 120 ° and three hemisphere is symmetrically fixedly installed
The forced snap lock mandrel of shape；In use, the locating dowel and the detent of the micro-fluidic chip are oriented to position is closed
Effect, three forced snap lock mandrels of hemispherical are tensioned by elastic reaction to the micro-fluidic chip MC.
The symmetrical structure or asymmetrical structure composition bifocal segment that focus is conjugated before and after the object lens and imaging len use
Imaging lens group, the object lens are made up of six lens altogether with imaging len, and the combination of the object lens and imaging len is as follows：
Object lens are made up of the first eyeglass, the second eyeglass and the 3rd eyeglass；The imaging len is by the 4th eyeglass, the 5th eyeglass and the 6th mirror
Piece forms；Wherein, it is saturating with the 3rd eyeglass, the 4th eyeglass to separately constitute two gluings with the 5th eyeglass for second eyeglass
Mirror, the parameter of each eyeglass are as shown in table 1：
Each lens parameters during the object lens of table 1 combine with imaging len
The semiconductor refrigerating light source module uses semiconductor refrigerating LED structure and leaded light device, the semiconductor refrigerating
LED structure includes LED luminescence chips, the first fin, semiconductor cooler, the second fin and fan；The leaded light device
The LED luminescence chips are connected, the LED luminescence chips connect first fin, the refrigeration of the semiconductor cooler
End is in close contact by heat-conducting silicone grease and the first fin SH1, and the radiating end of the semiconductor cooler passes through thermal conductive silicon
Fat is in close contact with second fin, and second fin connects the fan；Wherein, to the semiconductor cooler
Forward direction power supply can meet the temperature requirement of LED chip under hot environment in 0~25 DEG C of normal running conditions, to the semiconductor system
Cooler reverse power supply then meets the temperature requirement of 0~25 DEG C of normal running conditions of LED chip under low temperature environment；The leaded light device
Using lamp guide, optical fiber, eyeglass or speculum.
For the present invention due to taking above technical scheme, it has advantages below：1st, the present invention is due to using by 6 sections of primer sets
Into detection of nucleic acids molecular probe, detected for different indicator nucleic acid, 6 sections of design of primers are into different A, T, G, C base sequences
The nucleic acid fragment of row order and length, therefore with the specificity to the pathogen of different genera, virus, microorganism, fungi etc.
Detect identification function.2nd, the present invention, can be in miniflow due to being mixed using detection of nucleic acids reagent with analyzed nucleic acid samples
Trace nucleic acid samples molecule diagnostic application is realized on control chip MC, realizes the μ L of the reaction system of single index detection of nucleic acids≤1.5, inspection
Limit is surveyed to reach within 10 nucleic acid molecules copies.3rd, the present invention using the bath of submillimeter laminar air due to flowing mode of heating to micro-
Fluidic chip is heated, and is driven micro-fluidic chip rotational movement air flow by motor, is become heating-up temperature field
Uniformly, ensure that the temperature on micro-fluidic chip between reaction channel has good uniformity.4th, the present invention is due to using accurate
Driving stepper motor pulley drive structure, by optimizing driving current, gearratio and being loaded using spring card location structure solid
Determine micro-fluidic chip, realize rotating speed >=6000 rev/min of the micro-fluidic chip in high speed rotating centrifugal and low in high accuracy
Rotating speed≤2 rev/min during fast rotation sweep are simultaneously deposited, and meet micro-fluidic chip auto injection requirement, and use digital subdivision side
Method, by zero-in, realize high-precision 2 revs/min of rotation sweep detections of low speed.5th, the present invention using spring card due to being determined
The fixed micro-fluidic chip of bit architecture carrying, ensures that micro-fluidic chip does not take off during high speed rotating centrifugal >=6000 rev/min
Fall to fly out.6th, the present invention is meeting compared with high score due to using object lens and imaging len as parfocal symmetric double focal plane imaging structure
On the premise of resolution and phosphor collection efficiency reach the optical diffraction limit of system, system architecture is simplified, have compressed object image distance
From.7th, the present invention realizes hot environment (≤65 DEG C) or low temperature environment (>=-48 due to using semiconductor refrigerating LED structure
DEG C) under maintain 0~25 DEG C of LED chip normal working temperature, so as to ensure LED light source stable luminescence under high and low temperature environment.8、
The present invention is not only suitable for clinical hospitals, family, community medicine due to the power supply mode using net electricity, solar energy and battery compatibility
Unit uses, and meets that sea, island, natural calamity area, outdoor, the net electricity such as outlying mountain area are interrupted or net is electric obstructed special
Environment requirement.9th, the present invention carries out salt spray resistance due to using the active heat radiation sealed moisture-proof construction of housing, and to its surface
Corrosion treatment, moisture proof, the function of resisting salt fog corrosion are made it have, meet humidity 95%, salt fog 5mg/m3Marine climate bar
The requirement of part and natural calamity area environmental condition.In summary, the present invention can be widely applied to outdoor, marine, net electricity
Detection of nucleic acids under the special environment condition such as obstructed outlying mountain area and earthquake or typhoon disaster area.
Brief description of the drawings
Fig. 1 is the micro-fluidic chip nucleic acid paralleled detection systems structural representation of the present invention；
Fig. 2 is the structural representation of the micro-fluidic chip of the present invention；
Fig. 3 (a) is the spring card location structure schematic diagram of the present invention, and Fig. 3 (b) is the spring card positioning knot of the present invention
Structure fixes the view of micro-fluidic chip；
Fig. 4 is the structural representation of the bifocal segment imaging lens group of the present invention；
Fig. 5 is that the motor of the present invention controls the view of micro-fluidic chip motion by pulley drive structure；
Fig. 6 is the semiconductor refrigerating LED structure schematic diagram of the present invention；
Fig. 7 is the solar powered structural representation of the present invention.
Embodiment
Come to carry out the present invention detailed description below in conjunction with accompanying drawing.It should be appreciated, however, that accompanying drawing has been provided only more
Understand the present invention well, they should not be interpreted as limitation of the present invention.In the description of the invention, it is to be understood that
Term " first ", " second " etc. are only used for the purpose of description, and it is not intended that instruction or hint relative importance.
As shown in figure 1, the micro-fluidic chip nucleic acid paralleled detection systems of the present invention include micro-fluidic chip MC, spring card
Location structure DW, pulley drive structure PD, motor M, motion-control module MD, heating film HF, temperature control modules PID, display
Device LS, power supply ES, signal acquisition process device Pro, semiconductor refrigerating light source module PLED, condenser L3, exciter filter F1, two
To Look mirror D, speculum R, object lens L1, imaging len L2, transmitting optical filter F2, pinhole diaphragm PH and detector PMT.
Micro-fluidic chip MC is fixed by spring card location structure DW, and spring card location structure DW passes through belt wheel
Drive mechanism PD connections motor M, motor M connection motion-control module MD；Heating film HF, miniflow are set around micro-fluidic chip MC
The air layer of submillimeter, heating film HF connection temperature control modules PID are kept between control chip MC and heating film HF；Motion control
Module MD, temperature control modules PID and display LS are connected to signal acquisition process device Pro.Signal acquisition process device Pro will
Motion control signal sends to motion-control module MD, motion-control module MD and controls electricity according to the motion control signal received
Machine M, motor M drive micro-fluidic chip MC to rotate by pulley drive structure PD；Meanwhile signal acquisition process device Pro is by temperature
Control signal is delivered to temperature control modules PID, and temperature control modules PID controls heating according to the temperature control signals received
Film HF temperature.Power supply ES is used to be each being powered with single device in micro-fluidic chip nucleic acid detection system.
Signal acquisition process device Pro control semiconductor refrigerating light source module PLED transmitting exciting lights, semiconductor refrigerating light source
The exciting light of transmitting is transmitted into exciter filter F1 by module PLED after condenser L3 collimations, and exciter filter F1 is by exciting light
It is filtered into after quasi- monochromatic excitation light and is transmitted into speculum R through dichroscope D, the exciting light reflected through speculum R passes through object lens L1
Convergence irradiation micro-fluidic chip MC reaction channel, excites biological sample in micro-fluidic chip MC reaction channels to produce fluorescence.It is glimmering
Light is transmitted into speculum R after object lens L1 convergences are collimated into directional light, the fluorescent emission through speculum R reflections to dichroscope D,
Fluorescent emission through dichroscope D reflections is filtered off except after the exciting light in fluorescence, imaged lens L2 to optical filter F2 is launched
Pinhole diaphragm PH is converged to, the fluorescence after pinhole diaphragm PH removes veiling glare is received by detector PMT, and detector PMT will connect
The signal of receipts is sent to signal acquisition process device Pro generation real-time fluorescence detection signals, and show by display LS real-time glimmering
Light detecting signal.
In a preferred embodiment, as shown in Fig. 2 micro-fluidic chip MC includes a bottom plate, bottom plate centrally disposed one
Positioning round orifice DC, positioning round orifice DC edge set a detent DL.Set on the bottom plate around positioning round orifice DC
There is the microfluidic channel of one or more, every microfluidic channel includes a microchannel TL, if being provided with each microchannel TL
Dry reaction passage T, each microchannel TL one end set sample holes TH, each microchannel TL other end to set venthole EH,
A buffer pool BC is provided with microchannel TL before each venthole EH, for collecting surplus liquid, ensures that venthole EH is entering
There is no fluid leakage in sample, be advantageous to the sealing of venthole.The reaction channel of micro-fluidic chip is also anti-comprising positive quality control control
Passage and negative Quality Control control reaction passage are answered, validity instruction is carried out to nucleic acid specificity detection recognition result.Positive quality control
Control reaction passage is fixed with the standard quality-control product index nucleic acid probe that can produce fluorescence signal, negative Quality Control control reaction passage
The standard quality-control product index probe either blank probe of fluorescence signal can not be produced by being fixed with.
In a preferred embodiment, as shown in Fig. 3 (a), spring card location structure DE includes a positioning seat XZ, one
The locating dowel DWX and forced snap lock mandrel HPD of three hemisphericals.Positioning seat XZ centrally disposed one and belt wheel transmission mechanism PD is carried out
The locating dowel DWX being used cooperatively with detent DL, positioning is fixedly installed in the through hole XZK of connection, positioning seat XZ side
Divide equally the forced snap lock mandrel HPD of three hemisphericals of symmetrical fixed setting in 120 ° around seat XZ.As shown in Fig. 3 (b), Spring Card
Piece location structure DW is used to carry fixed micro-fluidic chip MC, in use, locating dowel DWX and micro-fluidic chip MC detent
DL is with position guide effect is closed, and the forced snap lock mandrel HPD of three hemisphericals is by elastic reaction to micro-fluidic chip MC
It is tensioned, ensures that micro-fluidic chip is not fallen off during high speed rotating centrifugal >=6000 rev/min and fly out.
In a preferred embodiment, object lens L1 and imaging len L2 uses bifocal segment imaging detection mode, object lens L1
The symmetrical structure or asymmetrical structure composition bifocal segment imaging lens group that focus is conjugated before and after being used with imaging len L2, make glimmering
Light collection efficiency close to the optical diffraction limit of system, for parallel optical transport or the limited angle of divergence transmit by centre.As shown in figure 4,
Object lens L1 and imaging len L2 is made up of 6 lens altogether, makes phosphor collection efficiency close to the optical diffraction limit of system, middle
Transmitted for parallel optical transport or the limited angle of divergence.Object lens L1 and imaging len L2 combination are as follows：Object lens L1 is by the first eyeglass
L11, the second eyeglass L12 and the 3rd eyeglass L13 compositions；Imaging len L2 is by the 4th eyeglass L14, the 5th eyeglass L15 and the 6th mirror
Piece L16 is formed.Wherein, the second eyeglass L12 and the 3rd eyeglass L13, the 4th eyeglass L14 and the 5th eyeglass L15 separately constitute two
Balsaming lens, the parameter of each eyeglass are as shown in table 1.Object lens L1 and imaging len L2 is made by optics cold working and optical coating
Complete, its trueness error scope is the light that the phosphor collection efficiency that ± 1.0mm, object lens L1 and imaging len L2 are combined reaches system
Learn diffraction limit, detection sensitivity >=10 nucleic acid molecules copy/index.
Each lens parameters during the object lens L1 of table 1 combines with imaging len L2
In a preferred embodiment, as shown in figure 5, motor M can use stepper motor DJ of the prior art, band
Wheel drive mechanism PD can use existing device；Wherein, stepper motor DJ output shaft is fixedly connected with driving wheel DL, driving wheel DL
It is fixedly installed on by tooth band PDD connections with follower BL, follower BL on spring card location structure DW positioning seat XZ.
During work, motion-control module MD drives micro-fluidic chip MC according to the motion control signal controlled motor M received, realizes high
Fast rotating centrifugal >=6000 rev/min and high-precision low speed rotation scan≤2 revs/min, meet that micro-fluidic chip MC enters automatically
Sample and the requirement of real-time fluorescent signals detection, wherein, motion-control module MD is the ripe device of prior art, herein not
Repeat again；In addition, motor M is not limited to stepper motor, motor or direct current generator can also be taken using waiing upon.
In a preferred embodiment, as shown in fig. 6, semiconductor refrigerating light source module PLED uses semiconductor refrigerating
LED structure and lamp guide GB, semiconductor refrigerating LED structure include LED luminescence chips LC, the first fin SH1, semiconductor refrigerating
Device PWT, the second fin SH2, fan FAN.Lamp guide GB connection LED luminescence chips LC, LED luminescence chip LC connections first dissipate
Backing SH1, semiconductor cooler PWT refrigeration end are in close contact by heat-conducting silicone grease and the first fin SH1, semiconductor refrigerating
Device PWT radiating end is in close contact by heat-conducting silicone grease and the second fin SH2, the second fin SH2 connection fans FAN.It is right
The positive power supplies of semiconductor cooler PWT can meet that LED chip is in 0~25 DEG C of normal running conditions under hot environment (≤65 DEG C)
Temperature requirement, LED chip 0 under low temperature environment (>=-48 DEG C) can be met if to semiconductor cooler PWT reverse power supply
The temperature requirement of~25 DEG C of normal running conditions.Wherein, the leaded light device such as optical fiber or eyeglass or speculum can also be used to replace
Lamp guide GB, carry out remote isolation and pass light, reducing heat radiation of the environment temperature on LED luminescence chips influences, and improves light source
Stability, meet the requirement of the high and low temperature environment of -48 DEG C~65 DEG C scopes.
In a preferred embodiment, power supply ES can use the power supply mode of net electricity, solar energy and battery compatibility.Net
Electricity can be connected by power supply adaptor and used；Battery can use lithium battery or chemical cell；As shown in fig. 7, solar energy supplies
Electric system includes battery CL, charging voltage regulator controllers CD and solar panel SB, battery CL pass through the Regulation Control that charges
Device CD connects with solar panel SB.
In a preferred embodiment, the fast parallel detecting system of micro-fluidic chip nucleic acid can use resisting salt fog corrosion
Surface treatment and the active heat radiation sealed moisture-proof construction of housing.
In a preferred embodiment, heating film HF and temperature control modules PID is responsible for ensureing micro-fluidic chip MC's
Temperature meets the condition requirement of nucleic acid amplification reaction, and temperature control modules PID is prior art maturation device, no longer superfluous herein
State.
In a preferred embodiment, signal acquisition process device Pro can pass through testing result wired or wireless logical
The mode of news carries out data transmission, and wire communication mode includes parallel port, serial ports, USB, Ethernet or 1394 etc., wireless telecommunications bag
Include bluetooth, WIFI or Zigbee etc..
In summary, the fast parallel detection method of micro-fluidic chip nucleic acid provided by the invention, comprises the following steps：
1st, according to some indicator nucleic acids to be detected, corresponding detection of nucleic acids is separately fixed at molecular probe micro-fluidic
The reaction channel bottom of chip.
Wherein, detection of nucleic acids is made up of with molecular probe 6 sections of primers, is fixed on micro-fluidic chip MC reaction channel bottom
Portion, detected for different indicator nucleic acids, 6 sections of primers are designed to the nucleic acid of different A, T, G, C base sequence orders and length
Fragment, there is the specific detection identification function to the pathogen of different genera, virus, microorganism, fungi etc.；
Detection of nucleic acids can be embedded in micro-fluidic with molecular probe with other oligosaccharide or low melting point bio-intermiscibility material
Chip MC reaction channel bottom, such as sugar alcohol, maltose, heated by portable detection system in 40 DEG C~90 DEG C temperature models
Enclose interior fusing and discharge primer, mixed with sample and reaction reagent, carry out nucleic acid amplification reaction.
6 sections of primers of detection of nucleic acids molecular probe are adapted to nucleic acid isothermal amplification molecule diagnostic analysis, and it is used to detect greatly
One group of specific nucleic acid fragment sequence design result of enterobacteria is：
Primer No.1-GGCATCGTGG TGATTGATGA；
Primer No.2-GGTTCGTTGG CAATACTCCA；
Primer No.3-TCTTTCGGCT TGTTGCCCGC CTGCTGTCGG CTTTAACCTC；
Primer No.4-TACAGCGAAG AGGCAGTCAA CGGGTTTTTG TCACGCGCTA TC；
Primer No.5-TTCGAAACCA ATGCCTAAAG A；
Primer No.6-GCGCACTTAC AGGCGATT.
2nd, the nucleic acid samples being analysed to are dissolved in detection of nucleic acids reagent, will dissolve in nucleic acid samples using pipettor
Detection of nucleic acids reagent inject corresponding microfluidic channel from each sample holes of micro-fluidic chip, it is micro- in specific implementation process
Fluidic chip MC sample holes are fitted close with pipettor gun head, by preset one section of air column in pipettor during sample introduction
Top, to ensure that micro-fluidic chip sample-adding terminates after extracting pipettor gun head, its sample holes does not have fluid leakage, is advantageous to sample introduction
The sealing in hole.
Wherein, detection of nucleic acids with reagent mainly by dNTPs, EvaGreen, DTT, BstDNA Polymerase Buffer,
Tris-HCl (25 DEG C of pH 8.8at), MgSO4, M-MLV reverse transcriptase, RNasin Plus, BstDNA
Polymerase, Betaine are formed, and trace nucleic acid samples molecule diagnostic application, single index can be realized on micro-fluidic chip
The μ L of the reaction system of detection of nucleic acids≤1.5, test limit reach within 10 nucleic acid molecules copies.
3rd, signal acquisition process device Pro realizes high speed rotating centrifugal by controlled motor MC and low speed rotation is scanned and deposited,
Signal acquisition process device Pro drives micro-fluidic chip MC to revolve at a high speed in the sample introduction stage by motion-control module MD controlled motors M
Turn, make nucleic acid samples in microfluidic channel by high speed rotating centrifugal automatically into micro-fluidic chip MC reaction channel,
After completing sample introduction, signal acquisition process device Pro by controlling same motor M to drive micro-fluidic chip MC low speed rotations to scan,
Realize to the fast parallel detection of multiple reaction channels on micro-fluidic chip；Signal acquisition process device Pro passes through temperature control mould
Block PID control heating film HF, the bath flowing heating of submillimeter laminar air is carried out to micro-fluidic chip MC, micro-fluidic chip MC is simultaneously
Rotated under motor M drivings, drive air flow so that the temperature on micro-fluidic chip between reaction channel is uniform；
Held according to practical application by temperature control modules PID control micro-fluidic chip MC a certain in 40 DEG C~90 DEG C temperature ranges
Temperature so that the primer of each reaction channel is released in micro-fluidic chip MC, and with nucleic acid samples and detection of nucleic acids reagent
Mixing, carries out nucleic acid amplification reaction under the conditions of isothermal duplication, realizes that trace nucleic acid samples molecule diagnoses.
4th, using light irradiation micro-fluidic chip is excited, the production under the exciting of exciting light of the nucleic acid samples in micro-fluidic chip is made
Raw fluorescence；Phosphor collection efficiency is set to reach optical diffraction limit by bifocal segment imaging optical system structure；Fluorescence is concentrated in
Produce analog signal on detector, it is glimmering in real time that caused analog signal is sent to signal acquisition process device Pro generations by detector
Light detecting signal, and real-time fluorescence detection signal curve is shown by display LS.
The various embodiments described above are merely to illustrate the present invention, wherein the structure of each part, connected mode and manufacture craft etc. are all
It can be varied from, every equivalents carried out on the basis of technical solution of the present invention and improvement, should not exclude
Outside protection scope of the present invention.
Claims (8)
1. a kind of fast parallel detection method of micro-fluidic chip nucleic acid, comprises the following steps：
1) one is set to include micro-fluidic chip, motor, exciting light, bifocal segment imaging lens group, detector, signal acquisition process
Device, temperature control modules, the micro-fluidic chip nucleic acid detection system of motion-control module and display；Wherein, micro-fluidic chip
Comprising more than one reaction channel, heating film is provided with around micro-fluidic chip, keeps sub- between micro-fluidic chip and heating film
The air layer of millimeter；
2) according to some indicator nucleic acids to be detected, corresponding detection of nucleic acids is separately fixed at micro-fluidic chip with molecular probe
Reaction channel bottom；
3) nucleic acid samples being analysed to are dissolved in detection of nucleic acids reagent, and the core of nucleic acid samples will have been dissolved in using pipettor
Sour detection reagent injects corresponding microfluidic channel from each sample holes of micro-fluidic chip；
4) signal acquisition process device realizes high speed rotating centrifugal with low speed rotation scanning simultaneously by moving control module for controlling motor
Deposit, signal acquisition process device drives micro-fluidic chip to rotate at a high speed in the sample introduction stage by controlled motor, makes in microfluidic channel
Nucleic acid samples respectively enterd automatically by high speed rotating centrifugal in the reaction channel of micro-fluidic chip；After completing sample introduction, signal
Acquisition Processor is scanned by the same motor driven micro-fluidic chip low speed rotation of moving control module for controlling, is realized to micro-fluidic
The fast parallel detection of multiple reaction channels on chip；Synchronous signal Acquisition Processor is controlled by temperature control modules and heated
Film, the bath flowing heating of submillimeter laminar air is carried out to micro-fluidic chip, and micro-fluidic chip rotates under motor driving simultaneously
Motion, drive air flow so that the temperature on micro-fluidic chip between reaction channel is uniform；Temperature is passed through according to practical application
Control module control micro-fluidic chip is maintained at a certain temperature in 40 DEG C~90 DEG C temperature ranges so that each in micro-fluidic chip
The primer of reaction channel is released, and is mixed with nucleic acid samples and detection of nucleic acids with reagent, is carried out under the conditions of isothermal duplication
Nucleic acid amplification reaction, realize that trace nucleic acid samples molecule diagnoses；Wherein, micro-fluidic chip is consolidated by spring card location structure
It is fixed, and rotating speed >=6000 rev/min by pulley drive structure under motor driving during high speed rotating centrifugal sample introduction；It is micro-fluidic
Chip carries out low speed rotation scanning, rotating speed≤2 rev/min of low speed rotation scanning, angle positioning under the driving of same motor
Accuracy error≤0.1 °, wherein, spring card location structure includes a positioning seat, a locating dowel and the forced elasticity of three hemisphericals
Lock shaft；The through hole that positioning seat centrally disposed one is attached with belt wheel mechanism, the fixed setting of positioning seat side are recessed with positioning
The locating dowel that groove is used cooperatively, divide equally in 120 ° around positioning seat and the forced elasticity of three hemisphericals is symmetrically fixedly installed
Lock shaft；In use, the detent of locating dowel and micro-fluidic chip, with position guide effect is closed, three hemisphericals are forced
Snap lock mandrel is tensioned by elastic reaction to micro-fluidic chip (MC)；
5) using light irradiation micro-fluidic chip is excited, the nucleic acid samples in micro-fluidic chip are made to be produced under the exciting of exciting light glimmering
Light, phosphor collection efficiency is reached optical diffraction limit by bifocal segment imaging lens group, be emitted through bifocal segment imaging lens group
Fluorescence be accumulated and produce analog signal on the detector, caused analog signal is sent to signal acquisition process device by detector
Real-time fluorescence detection signal is generated, and real-time fluorescence detection signal is shown by display.
A kind of 2. fast parallel detection method of micro-fluidic chip nucleic acid as claimed in claim 1, it is characterised in that：The step
2) amplifying nucleic acid detection is fixed on the reaction channel bottom on micro-fluidic chip with molecular probe by adsorption, or using fine jade
The Bc material embedding of lipolysaccharide or oligosaccharide or fusing point in the range of 40 DEG C~90 DEG C is fixed on the anti-of micro-fluidic chip
Answer channel bottom.
A kind of 3. fast parallel detection method of micro-fluidic chip nucleic acid as claimed in claim 1, it is characterised in that：Detection of nucleic acids
It is made up of 6 sections of primers with molecular probe, is detected for different indicator nucleic acid, 6 sections of design of primers is into different A, T, G, C base sequences
The nucleic acid fragment of row order and length；Detection of nucleic acids is with reagent mainly by dNTPs, EvaGreen, DTT, BstDNA
25 DEG C of Polymerase Buffer, pH 8.8at Tris-HCl, MgSO4, M-MLV reverse transcriptase,
RNasin Plus, BstDNA Polymerase and Betaine are formed, the μ L of overall reaction system≤1.5, wherein, dNTPs is deoxidation
Ribonucleotide triphosphate, DTT are dithiothreitol (DTT), and Polymerase Buffer are polymerase buffer, 25 DEG C of pH 8.8at
For at 8.8,25 DEG C of acid-base value, Tris-HCl is Tri(Hydroxymethyl) Amino Methane Hydrochloride, MgSO4For magnesium sulfate, reverse
Transcriptase is reverse transcriptase, RNasin Plus be RNase inhibitor+, Polymerase is polymerase, Betaine
For glycine betaine.
A kind of 4. fast parallel detection method of micro-fluidic chip nucleic acid as claimed in claim 2, it is characterised in that：Detection of nucleic acids
It is made up of 6 sections of primers with molecular probe, is detected for different indicator nucleic acid, 6 sections of design of primers is into different A, T, G, C base sequences
The nucleic acid fragment of row order and length；Detection of nucleic acids is with reagent mainly by dNTPs, EvaGreen, DTT, BstDNA
25 DEG C of Polymerase Buffer, pH 8.8at Tris-HCl, MgSO4, M-MLV reverse transcriptase,
RNasin Plus, BstDNA Polymerase and Betaine are formed, the μ L of overall reaction system≤1.5, wherein, dNTPs is deoxidation
Ribonucleotide triphosphate, DTT are dithiothreitol (DTT), and Polymerase Buffer are polymerase buffer, 25 DEG C of pH 8.8at
For at 8.8,25 DEG C of acid-base value, Tris-HCl is Tri(Hydroxymethyl) Amino Methane Hydrochloride, MgSO4For magnesium sulfate, reverse
Transcriptase is reverse transcriptase, RNasin Plus be RNase inhibitor+, Polymerase is polymerase, Betaine
For glycine betaine.
5. a kind of fast parallel detection method of micro-fluidic chip nucleic acid as described in any one of Claims 1 to 4, its feature exist
In：The step 3) will dissolve in the detection of nucleic acids reagent of nucleic acid samples from each sample introduction of micro-fluidic chip using pipettor
The detailed process that corresponding microfluidic channel is injected in hole is：Each sample holes on micro-fluidic chip are closely matched somebody with somebody with pipettor gun head
Close, preset one section of air column is on the top of pipettor during sample introduction.
6. a kind of system for realizing the fast parallel detection method of micro-fluidic chip nucleic acid as described in any one of Claims 1 to 5, its
It is characterised by：It include micro-fluidic chip, spring card location structure, pulley drive structure, motor, motion-control module, plus
Hotting mask, temperature control modules, display, power supply, signal acquisition process device, semiconductor refrigerating light source module, condenser, excite filter
Mating plate, dichroscope, speculum, object lens, imaging len, transmitting optical filter, pinhole diaphragm and detector；The micro-fluidic chip
It is fixed by the spring card location structure, the spring card location structure is connected by the pulley drive structure
The motor, motion-control module described in the motor connection；The heating film is set around the micro-fluidic chip, it is described micro-
The air layer of submillimeter is kept between fluidic chip and the heating film, the heating film connects the temperature control modules；Institute
State motion-control module, temperature control modules and display and be connected to the signal acquisition process device；At the signal acquisition
Reason device sends motion control signal to the motion-control module, and the motion-control module is according to the motion control received
Signal controls the motor, and the motor drives the micro-fluidic chip to rotate by the pulley drive structure；It is meanwhile described
Temperature control signals are delivered to the temperature control modules by signal acquisition process device, and the temperature control modules are according to receiving
Temperature control signals control the heating film, the micro-fluidic chip is reached the temperature that biological sample reaction requires；It is described
Power supply is used to be powered for each electricity consumption device, and the power supply (ES) is using the supplier of electricity that net is electric, solar energy and battery are compatible
Formula；
The signal acquisition process device controls the semiconductor refrigerating light source module transmitting exciting light, the semiconductor refrigerating light source
The exciting light of transmitting is transmitted into the exciter filter by module after condenser collimation, and the exciter filter will excite
Light is filtered into after quasi- monochromatic excitation light and is transmitted into speculum through the dichroscope, and the exciting light reflected through the speculum passes through
The reaction channel of the micro-fluidic chip is irradiated in the object lens convergence, excites biological sample in the micro-fluidic chip reaction channel
Produce fluorescence；The fluorescence is transmitted into the speculum after object lens convergence is collimated into directional light, anti-through the speculum
To the dichroscope, the fluorescent emission through dichroscope reflection filters off the fluorescent emission penetrated to the transmitting optical filter
After the exciting light in fluorescence, the pinhole diaphragm is converged to through the imaging len, veiling glare is removed through the pinhole diaphragm
Fluorescence afterwards is received by the detector, and the signal for detecting reception is sent to the signal acquisition process device and given birth to by the detector
Real-time fluorescence detection signal is shown into real-time fluorescence detection signal, and by the display.
7. system as claimed in claim 6, it is characterised in that：The micro-fluidic chip includes a bottom plate, the bottom plate center
One positioning round orifice is set, and the edge of the positioning round orifice sets a detent；On the bottom plate around the positioning round orifice
It is provided with the microfluidic channel of one or more, every microfluidic channel includes a microchannel, on each microchannel
Some reaction channels are provided with, one end of each microchannel sets a sample holes, and the other end of each microchannel is set
Put a venthole, buffer pool is provided with the microchannel before each venthole；Wherein, the reaction channel is also comprising the positive
Quality Control control reaction passage and negative Quality Control control reaction passage, the positive quality control control reaction passage be fixed with can produce it is glimmering
The standard quality-control product index nucleic acid probe of optical signal, the negative Quality Control control reaction passage, which is fixed with, can not produce fluorescence signal
Standard quality-control product index probe either blank probe.
8. system as claimed in claims 6 or 7, it is characterised in that：The semiconductor refrigerating light source module uses semiconductor system
Cold LED structure and leaded light device, the semiconductor refrigerating LED structure include LED luminescence chips, the first fin, semiconductor system
Cooler, the second fin and fan；The leaded light device connects the LED luminescence chips, described in the LED luminescence chips connection
First fin, the refrigeration end of the semiconductor cooler are in close contact by heat-conducting silicone grease and first fin (SH1),
The radiating end of the semiconductor cooler is in close contact by heat-conducting silicone grease and second fin, and second fin connects
Connect the fan；Wherein, power supply positive to the semiconductor cooler can meet under hot environment LED chip at 0~25 DEG C just
The temperature requirement of normal condition of work, then meets 0~25 DEG C of LED chip under low temperature environment to the semiconductor cooler reverse power supply
The temperature requirement of normal running conditions；The leaded light device uses lamp guide, optical fiber, eyeglass or speculum.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510078655.5A CN104630373B (en) | 2015-02-13 | 2015-02-13 | A kind of fast parallel detection method of micro-fluidic chip nucleic acid and system |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510078655.5A CN104630373B (en) | 2015-02-13 | 2015-02-13 | A kind of fast parallel detection method of micro-fluidic chip nucleic acid and system |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN104630373A CN104630373A (en) | 2015-05-20 |
| CN104630373B true CN104630373B (en) | 2017-11-21 |
Family
ID=53209617
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201510078655.5A Active CN104630373B (en) | 2015-02-13 | 2015-02-13 | A kind of fast parallel detection method of micro-fluidic chip nucleic acid and system |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN104630373B (en) |
Families Citing this family (29)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2016500002A (en) * | 2012-10-05 | 2016-01-07 | カリフォルニア インスティテュート オブ テクノロジー | Methods and systems for microfluidic imaging and analysis |
| CN107034266A (en) * | 2016-01-29 | 2017-08-11 | 清华大学 | For detecting that the primer of wound infection pathogen is combined and integrating device |
| WO2017139447A1 (en) * | 2016-02-10 | 2017-08-17 | Coyote Bioscience Usa Inc. | Methods and systems for analyzing nucleic acids |
| CN106085842B (en) * | 2016-06-16 | 2018-06-29 | 清华大学 | A kind of high throughput micro-fluidic chip nucleic acid amplification assays detecting system |
| CN106222069B (en) * | 2016-08-31 | 2018-11-09 | 上海科源电子科技有限公司 | A kind of disc-type chip PCR instrument and detection method |
| US10850281B2 (en) | 2016-09-12 | 2020-12-01 | Delta Electronics Int'l (Singapore) Pte Ltd | Nucleic acid analysis apparatus |
| CN107815408B (en) * | 2016-09-12 | 2021-11-09 | 台达电子国际（新加坡）私人有限公司 | Nucleic acid analysis device using isothermal amplification method |
| US11478791B2 (en) | 2016-09-12 | 2022-10-25 | Delta Electronics Int'l (Singapore) Pte Ltd | Flow control and processing cartridge |
| US11376581B2 (en) | 2016-09-12 | 2022-07-05 | Delta Electronics Int'l (Singapore) Pte Ltd | Flow control and processing cartridge |
| US11426735B2 (en) | 2016-09-12 | 2022-08-30 | Delta Electronics Int'l (Singapore) Pte Ltd | Nucleic acid analysis apparatus |
| CN106520517B (en) * | 2016-10-09 | 2019-06-28 | 上海交通大学医学院附属仁济医院 | A kind of portable micro-fluidic chip LAMP visual detector and its detection method |
| CN106979940A (en) * | 2017-04-17 | 2017-07-25 | 浙江大学 | A kind of device and method for carrying out nucleic acid amplification detection |
| CN107576639A (en) * | 2017-08-28 | 2018-01-12 | 博奥生物集团有限公司 | Portable fully integrated DNA spot examines micro-full analytical system light path |
| CN107807236A (en) * | 2017-10-31 | 2018-03-16 | 南京先进激光技术研究院 | One-color fluorescence detection means |
| CN108732370B (en) * | 2018-03-28 | 2021-07-09 | 无锡科智达科技有限公司 | Reagent averaging method |
| CN108277149B (en) * | 2018-06-04 | 2018-09-25 | 上海速创诊断产品有限公司 | Nucleic acid detection apparatus, method and system |
| CN108760686B (en) * | 2018-08-07 | 2024-05-14 | 天津诺迈科技有限公司 | Micro-fluidic chip for detecting turbidimetry and biochemical immunity machine using same |
| CN108913599B (en) * | 2018-08-10 | 2021-08-17 | 清华大学 | Long-time-range multimode information detection method and system for in-situ culture of living cells |
| CN111175289B (en) * | 2018-11-09 | 2022-06-28 | 广州中国科学院先进技术研究所 | Multi-functional quick water quality testing system based on multispectral imaging and micro-fluidic technique |
| CN109520987A (en) * | 2018-12-19 | 2019-03-26 | 苏州汶颢微流控技术股份有限公司 | ATP fluorescence detector |
| CN110373312B (en) * | 2019-08-22 | 2022-11-22 | 深圳市芯思微生物科技有限公司 | Constant-temperature amplification nucleic acid detection device and detection method |
| CN110846219B (en) * | 2019-11-14 | 2021-06-29 | 清华大学 | Optical fiber sensing micro-fluidic chip nucleic acid amplification in-situ real-time detection system and method |
| CN111157508B (en) * | 2020-03-06 | 2022-09-06 | 成都博奥晶芯生物科技有限公司 | Continuous acquisition method for fluorescence data of microfluidic chip |
| CN113832220A (en) * | 2020-06-24 | 2021-12-24 | 上海驷格生物科技有限公司 | PCR fluorescent nucleic acid detection method and application thereof |
| CN111748463A (en) * | 2020-08-07 | 2020-10-09 | 天津智善生物科技有限公司 | Amplification device applied to nucleic acid detection and detection method based on amplification device |
| CN112342276B (en) * | 2020-11-09 | 2024-05-24 | 成都博奥独立医学实验室有限公司 | Method for detecting micro-fluidic chip by using nucleic acid secondary structure non-specific fluorescence adsorption |
| CN112625895B (en) * | 2020-11-30 | 2022-12-23 | 清华大学深圳国际研究生院 | Micro-fluidic chip control device |
| CN113483657B (en) * | 2021-05-26 | 2023-09-29 | 广州万孚倍特生物技术有限公司 | Data acquisition method, detection turntable, device, computer equipment and storage medium |
| CN114112864A (en) * | 2021-11-22 | 2022-03-01 | 西北大学 | Counting, sorting and speed measuring system and method for biological samples and storage medium |
Family Cites Families (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN100535644C (en) * | 2007-09-21 | 2009-09-02 | 博奥生物有限公司 | Micro-nano system fluid chip detection system and detection method |
| CN102369443B (en) * | 2009-03-31 | 2015-08-19 | 凸版印刷株式会社 | The manufacture method of sample analysis chip, the sample analyzer adopting this sample analysis chip and sample analyzing method and sample analysis chip |
| JP5495104B2 (en) * | 2009-10-16 | 2014-05-21 | ウシオ電機株式会社 | Inspection equipment using chips |
| CN102253022B (en) * | 2011-05-04 | 2013-04-24 | 博奥生物有限公司 | Digital signal processing method for detecting microfluidic chip and applied detection device |
-
2015
- 2015-02-13 CN CN201510078655.5A patent/CN104630373B/en active Active
Also Published As
| Publication number | Publication date |
|---|---|
| CN104630373A (en) | 2015-05-20 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN104630373B (en) | A kind of fast parallel detection method of micro-fluidic chip nucleic acid and system | |
| CN106085842B (en) | A kind of high throughput micro-fluidic chip nucleic acid amplification assays detecting system | |
| CN101460953B (en) | Systems and devices for sequence by synthesis analysis | |
| CN100535644C (en) | Micro-nano system fluid chip detection system and detection method | |
| CN104267009B (en) | Six color real-time fluorescence quantitative PCR analyzers | |
| CN106906137A (en) | A kind of high-throughput nucleic acid analysis device based on micro-fluidic chip | |
| CN107805597A (en) | Gene detection system and detection method based on micro-fluidic chip | |
| KR20170024827A (en) | The Quantitative PCR Cartridge with Microchannel-Film Reactor, Nucleic Acid Extraction Module and qPCR Reagents Module, and The Rapid qPCR System Using the Same | |
| CN202898426U (en) | Space-oriented spiral micro-fluidic PCR (Polymerase Chain Reaction) real-time fluorescence detection system | |
| CN103667012B (en) | Microfluidic PCR (Polymerase Chain Reaction) chip fluorescence fluid detection device based on CCD (Charge Coupled Device) image sensor | |
| CN102286358A (en) | Microfluidic control chip for realizing PCR (Polymerase Chain Reaction) and real-time PCR virus quick detection device | |
| CN108642158A (en) | A kind of PCR real-time fluorescence detection systems of multichannel point detection | |
| CN107159328A (en) | Sample analysis system | |
| WO2021093622A1 (en) | Optical fiber sensing microfluidic chip nucleic acid amplification in-situ real-time detection system and method | |
| CN102618439A (en) | Deoxyribonucleic acid (DNA) fragment amplification and quantitative detection system based on closed reactors | |
| Jie et al. | Portable and battery-powered PCR device for DNA amplification and fluorescence detection | |
| CN110452803A (en) | A kind of nucleic acid rapid amplifying detection method and device | |
| CN110714049B (en) | Microbial sensor for detecting biomarkers, detection method, culture and detection chip and detection system | |
| CN106353295A (en) | Protein heat stability analysis meter | |
| Huang et al. | Ultra-fast, sensitive and low-cost real-time PCR system for nucleic acid detection | |
| CN219907656U (en) | Fluorescence detection mechanism for molecular diagnosis equipment | |
| AU2020100366A4 (en) | Gene detection system and method based on microfluidic chip | |
| CN212581870U (en) | System for extracting, purifying and amplifying nucleic acid | |
| CN110975960B (en) | Composite PCR system with dPCR and qPCR functions and control method | |
| CN113621475A (en) | Method and system for extracting, purifying and amplifying nucleic acid |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |