CN105802848A - Recombinase-mediated isothermal nucleic acid amplification reaction real-time detection apparatus - Google Patents
Recombinase-mediated isothermal nucleic acid amplification reaction real-time detection apparatus Download PDFInfo
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- CN105802848A CN105802848A CN201410717966.7A CN201410717966A CN105802848A CN 105802848 A CN105802848 A CN 105802848A CN 201410717966 A CN201410717966 A CN 201410717966A CN 105802848 A CN105802848 A CN 105802848A
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Abstract
The present invention discloses a recombinase-mediated isothermal nucleic acid amplification reaction real-time detection apparatus, which includes a microprocessor, a photovoltaic module, a reaction isothermal control system, a reaction time-control module, a vibration module and a sample loading reaction chamber. The microprocessor includes a temperature microcontroller, a time microcontroller and a photovoltaic module microcontroller. The reaction isothermal control system is based on the information received from a temperature sensor, and the temperature microcontroller uses a PID algorithm, and is connected to a PWM control circuit to adjust and control the temperature of a heater, so as to reach isothermal control. The apparatus further comprises a communication interface interactive with one PC or a plurality of other intelligent devices. The apparatus can realize real-time fast detection of nucleic acid amplification at a temperature of 25-45 DEG C 5 within 30 min, has the characteristics of high detection sensitivity, simple operation, and improved efficiency of nucleic acid amplification detection, and can be applied to a nucleic acid molecule laboratory research, medical diagnostics and on-site testing.
Description
Technical field
The present invention relates to a kind of recombinase-mediated isothermal nucleic acid amplification reaction real-time detection apparatus, specifically equity isothermal nucleic acid expands
Volume increase thing carries out Real_time quantitative detection.
Background technology
In biological study and molecular diagnosis, DNA cloning is a kind of basic and necessary technological means, most common of which
Be to utilize polymerase chain reaction (English full name: Polymerase Chain Reaction is called for short PCR) to carry out nucleic acid
Amplification in vitro, this technology in nineteen eighty-three by American scientist PE (Perkin Elmer Po Jin-Ai Ermo) company
The Dr.Mullis invention in heredity portion, due to round pcr in theoretical and application across significance of times, therefore
Mullis obtains chemical Nobel Prize in 1993, be the most widely used in modernization agricultural and medical science and
The fields such as food industry.By this technology, it is possible to achieve the efficient amplification of trace dna, by minimal amount of specific nucleic acid
Acid molecules expands the level that instrument can detect.Round pcr nucleic acid amplification divides three fundamental reaction steps, respectively
It is degeneration, annealing (renaturation) and extension；Particularly as follows: 1. template DNA degeneration: template DNA is heated to 90-95 DEG C
After certain time, make template DNA double-strand or through PCR amplification formed double-stranded DNA untwist, make strand, with
Just it is combined with primer, prepares for lower whorl reaction；2. template DNA and the annealing (renaturation) of primer: template DNA
After heated degeneration becomes strand, temperature is down to 55-60 DEG C, and primer combines with the complementary series pairing of template DNA strand；
3. the extension of primer: DNA profiling-primer conjugate is under the effect of archaeal dna polymerase, in 70-75 DEG C, with dNTP
For reaction raw materials, target sequence is template, by base pairing and semiconservative replication principle, synthesizes a new and template DNA
The semiconservative replication chain that chain is complementary, repetitive cycling degeneration-annealing extends three processes, so that it may obtain more " partly reservation
Replicate chain ", and this new chain can become the template of circulation next time.Often completing a circulation to need 2-4 minute, 2-3 is little
Genes of interest amplification is amplified millions of times by Shi Caineng.
In order to detect whether target gene Successful amplification, be developed the multiple method for detecting amplified production and
Instrument, traditional method is to use agarose gel electrophoresis to detect, and this method can only carry out one to amplified production
Analyze qualitatively, it is impossible to carry out quantitatively, can not nucleic acid amplification being monitored in real time to amplification template, and at low concentration
Easily producing false negative during amplification, sensitivity is low, is therefore constantly developing the detection method of pcr amplification product always
And renewal, up to the present, PCR detecting instrument has evolved to the 7th generation quantitative fluorescent PCR (realtime
Fluorescence quantitative PCR, RTFQ PCR)；This analytical tool be at first 1996 by the U.S.
Applied Biosystems company releases, and by fluorescent dye or fluorescent labeling specific probe, enters PCR primer
Line flag is followed the tracks of, real time and on line monitoring course of reaction, and reaction combines corresponding software after terminating and is analyzed product, and
Utilize the standard curve measured that the initial concentration of testing sample template is carried out quantitative Analysis, it is achieved quantitative analysis；But its
Design principle is also that the amplified reaction feature according to round pcr carries out particular design exploitation, needs in reaction temperature
90-95 DEG C of degeneration, 55-60 DEG C of annealing (renaturation), 70-75 DEG C of extension three phases, and through some amplification cycles ability
Completing, detecting instrument design complexity, instrument price is expensive, and needs to consume a large amount of electric power, is only applicable to there is supply of electric power
Laboratory, operating process require height, it is desirable to have professional and technical personnel support, the cost of the most this instrument and application model
Enclose all by a definite limitation.
The most nearly ten years, the DNA cloning technology in simulation microbial body is quietly risen, and makes nucleic acid in vitro amplification become more
Adding simple and convenient, such as TMA technology (nucleic acid amplification technologies of transcriptive intermediate), SDA technology, (strand displacement nucleic acid expands
Increasing technology), LAMP (ring mediation nucleic acid amplification technologies), HDA (unwindase dependence isothermal amplification), RAA (weight
Group enzyme mediated isothermal amplification) etc. all can DNA amplification under isothermal conditions.
The present invention be directed to recombinase-mediated isothermal amplification and invent for real time nucleic acid detection amplified production
Quantitative testing device.Recombinase-mediated isothermal amplification can realize under lower isothermy (25-45 DEG C), more
Fast nucleic acid amplification, process is easy, and the present invention utilizes this response characteristic of this technology to design, it is achieved in real time
The device of detection by quantitative amplified production.
This detection device have highly sensitive, volume is little, the advantage such as fast and convenient.With the real-time PCR rate of exchange
Lattice are low, and the reaction temperature of instrument controlling is low, it is not necessary to a large amount of electric power, easy and simple to handle, it is not necessary to professional and technical personnel operates
Supporting, the detection time is greatly shortened, typically can be obtained by testing result in 5-30 minute, makes nucleic acid amplification technologies letter
Dan Hua, and be conducive to the application of this type of technology wider scope, can apply to nucleic acid molecules laboratory research, medical diagnosis
With fields such as Site Detections.
Summary of the invention
It is an object of the invention to provide a kind of Real_time quantitative detection dress for measuring recombinase-mediated isothermal nucleic acid amplification product
Put, have highly sensitive, reaction control temperature is low, automaticity is high, operating process simply, quickly letter
Just, low cost and other advantages, it is achieved the quick detection to nucleic acid amplification product.
Recombinase-mediated isothermal nucleic acid amplification process temperature controls at 25-45 DEG C, and optimum reacting time is 37 DEG C, with existing
Any nucleic acid amplification reaction ratio, reaction temperature is lower, reaction efficiency faster, in order to coordinate this nucleic acid amplification skill
The detection in real time of amplified production has been invented this device by art.
Recombinase-mediated isothermal amplification is that one utilizes RecQ albumen, UvsY albumen and UvsX albumen waiting
Under the conditions of temperature (25-45 DEG C), optimum reacting time is 37 DEG C, makes template double-strand untwist, stablizes strand and make primer and template
Between occur chain replace such that it is able to substitute normal PCR degeneration (94 DEG C) and anneal (50-60 DEG C) step, meanwhile,
Nucleic acid under the catalysis of archaeal dna polymerase, at the bar that single strand binding protein SSB and condensing agent Polyethylene Glycol exist
Synthetic product under part, and can constantly repeat this process, in 5-30 minute, just can realize the efficient amplification of nucleic acid.
In principle, the reaction of recombinase-mediated isothermal nucleic acid amplification is an exponential amplification, and amplified reaction is only imitated with time and amplification
Rate is correlated with；This is relevant with period and amplification efficiency to the amplification of PCR is different.
In order to quickly enable amplification DNA out be detected and improve detection sensitivity, at recombinase-mediated isothermal
A specific fluorescent probe is added while nucleic acid amplification reaction reagent adds pair for amplification target gene primer,
This probe is an oligonucleotide, and two ends one reporter fluorescence group of labelling and a quenching fluorescence group respectively, probe is complete
Time, the fluorescence signal that reporter group is launched is quenched group absorptions；When target gene Successful amplification, the 5 ' reports held
Group splits away off along with the hydrolysis of probe, no longer with quenching group generation energy transmission effect, it is thus possible to send fluorescence,
Often one DNA of amplification, just has a fluorescence molecule to be formed, it is achieved the accumulation of fluorescence signal and amplified production are formed completely
Synchronizing, fluorescence volume power is directly proportional to amplified production；Therefore, there is one-to-one relationship between fluorescence volume and amplified production,
The detection to amplified production just can be realized, here it is the present invention's realizes principle by fluorescence volume is detected；
Concrete implementation is to utilize specific excitation source to excite specific fluorescent probe, produces specific fluorescence,
By fluorescent collecting and carry out opto-electronic conversion by photo-detector again, obtain the photosignal that can gather and process, microprocessor
Processing after receiving signal, the signal after processing shows and i.e. obtains real-time amplification curve on PC display screen；Further according to
The standard curve demarcated in advance can be obtained by the initial content of test sample by analyzing computed in software.
According to above principle, invent a kind of device for the detection of recombinase-mediated isothermal nucleic acid amplification product, comprise: a.
One can carry out Treatment Analysis and the microprocessor feeding back the unit controlled to the data gathered；
B. one can realize temperature controlling range at 25-45 DEG C, the reaction thermostatic control system of precision ± 0.1 DEG C；
C. example reaction room is loaded；
D. one group includes LED light source, excites the optical-electric module of Optical devices, transmitting optics and photo-detector；
E. the time microcontroller response time being controlled；
F. a shock module；
G. comprise at least one communication interface, can interact with host computer；Host computer can be PC or other intelligent apparatus；
By above module and unit, it is achieved to response time, reaction temperature, reaction shake control, it is achieved optical telecommunications
Number gather and conversion；And gathered by microprocessor and process, on host computer, finally show the real-time signal of amplified reaction
And draw out instant amplifying curve, complete the detection to amplified production.
Wherein microprocessor includes at least temperature microcontroller, response time microcontroller, optical-electric module microcontroller, and
There is data acquisition and process function；Temperature control, reaction can be carried out according to user by host computer operating system
Time controls, reaction shake control, LED light source control and can carry out signals collecting and process.
Reaction thermostatic control system by temperature microcontroller, PID controller, pwm control circuit, heater, heater and
Auxiliary device control circuitry and temperature sensor composition；Heater is the heater of metal or ceramic material, comprises
Having heaters and auxiliary device control circuitry and least one set metal fin and fan, and reception temperature microcontroller can be utilized
The instruction of device feedback is automatically obtained temperature regulation, it is achieved temperature controlling range 25-45 DEG C, precision ± 0.1 DEG C；
Temperature sensor has collecting temperature and is converted into the digital data transmission merit to temperature microcontroller by signal collected
Energy.Temperature microcontroller can be according to the signal of temperature sensor feedback by entering pwm control circuit after PID controller computing
Row controls, and if temperature is less than the reaction temperature set, then makes heater energising heat up；If temperature is higher than the reaction temperature set
Degree, then power-off starting fan are lowered the temperature by fin, it is achieved thermostatic control.
Load example reaction room and can carry out multiple sample test simultaneously, it is also possible to carry out single sample test；Sample energy can
To be designed according to realizing needs.
Optical-electric module includes LED light source, excites Optical devices and optical detection system.
Optical devices are excited to be made up of one or more groups lens, filter lens, dichroscope, reflecting mirror.
Optical detection system is made up of transmitting optics and photomultiplier tube；
Transmitting optics includes one or more groups lens, filter lens, dichroscope, reflecting mirror；
Photo-detector receives from the reacted fluorescence signal of sample, is believed by the photomultiplier tube fluorescence to receiving
Number it is amplified and opto-electronic conversion, becomes the discernible signal of telecommunication, feed back to microprocessor.
Shock module is formed with vibrations motor driver by shaking motor；After sample completes the preheating time that user sets
Time microcontroller, according to the Automatic Program starting shock motor driver set, makes vibrations motor shake response sample
Dynamic mixing, accelerates response speed.
Vibrations motor is connected with response sample room, when shaking motor operations, response sample can produce the effect of vibrations.
Microcontroller major control response time in response time and vibrations initiate, dwell time, and user can pass through host computer
On operating system response time and vibrations initial time are configured, it is possible to according to actual reaction to data acquisition
Time interval is configured；User can arrange result by the display screen on host computer is visible in detail.
User can set the response time of this detection device, reaction temperature, data acquisition by the operating system on host computer
Time interval, vibrations initial time and dwell time.
Display system on host computer is for showing between response time that user sets, reaction temperature, data acquisition time
Every, and immediate reaction time and reaction temperature can be shown；
Remaining with communication interface on this detection device, communication interface can be from including serial ports, USB, bluetooth, Wifi or its group
Conjunction is selected, it is also possible to retain multiple interface.
This detection device using method comprises the following steps:
A. external power supply or built-in power, opening device are connected；
B. set up between described detection device and host computer by communication interface and communicate；
C. by the operating system of host computer arrange detection the response time of device, reaction temperature, vibrations initial time and
The response parameter such as dwell time, data collection interval；
D. recombinase-mediated isothermal nucleic acid amplification reaction reagent is put into reative cell, starts the ginseng that detection program is set by user
Number reacts；
E. the signal of telecommunication that reaction after starting detects optical detection system is sent to microprocessor, and host computer calls these data
And analyze and process.
Above operating process user can create, revise or delete the reaction of detection device by the operating system of host computer
Time, reaction temperature, vibrations initial time and dwell time, data collection interval, these parameters are for recombinase
It is provided with default value in mediated isothermal amplification device, but user can these reaction conditions self-defined and data
Acquisition time is spaced.
User can demonstrate the real-time signal of amplified reaction by host computer and thus draw out instant amplifying curve.
Accompanying drawing explanation
Fig. 1 is the theory structure schematic diagram of the present invention；Figure carries in 101-PC machine or other processing means, i.e. description
The host computer arrived；102-communication interface (serial/bluetooth/USB interface/Wifi)；103-microprocessor；104-temperature microcontroller
Device；105-heater；106-temperature sensor；107-time microcontroller；108-shock module；109-LED light source；
110-excites Optical devices；111-photo-detector；112-transmitting optics；113-example reaction room；114-optical-electric module
Microcontroller；115-set of cells or external power supply；
Fig. 2 is the principle schematic that the present invention reacts an embodiment of thermostatic control system；201-0-24V power supply in figure；
104-temperature microcontroller (P89V51)；204-PWM control circuit；105-heater；106 temperature sensors (DS18B20)；
103-microprocessor；
Fig. 3 is the optical-electric module principle schematic of one embodiment of the invention；109-LED light source in figure；302-lens；
303-filter lens (exciting light)；304-dichroscope (exciting light)；305-reflecting mirror (exciting light)；306-lens (optically focused)；307-
Sample；308-lens；309-reflecting mirror (transmitting light)；310-dichroscope (transmitting light)；311-filter lens (transmitting light)；
312-lens (optically focused)；111-photo-detector；114-optical-electric module microcontroller；103-microprocessor；
Fig. 4 is the device appearance schematic diagram of the embodiment of the present invention；401-main frame (including microprocessor 103) in figure；402-shakes
Dynamic motor；105-heater；113-example reaction room；405-reacts chamber cap；406-reacting hole；102-communication interface；408-
Liquid crystal touch screen；409-on and off switch；410-external power interface；
Fig. 5 is the amplification figure in embodiment to soy bean DNA；Figure is the data of the real time measure show without analyzing software
Process.Amplification curve under wherein L9, L5, L4, L3, L2, L1 are target amplification DNA variable concentrations respectively,
NTC is negative control；
Detailed description of the invention
Below in conjunction with embodiment, the present invention is described in detail.
The present invention relates to a kind of device for the detection of recombinase-mediated isothermal nucleic acid amplification product, comprise:
A. one can carry out Treatment Analysis and the microprocessor 103 feeding back the unit controlled to the data gathered；
B. one can realize temperature controlling range at 25-45 DEG C, and the reaction thermostatic control system of precision ± 0.1 DEG C, such as Fig. 2 institute
Show；
C. the reative cell 113 loading sample in the present embodiment has 8 example reaction holes；
D. one group includes LED light source 109, excites Optical devices 110, transmitting optics 112, photo-detector 111 and photoelectricity
The optical-electric module of module microcontroller 111；
E. a time microcontroller 107 response time being controlled and shock module 108；
F. use at least one USB as communication interface 102, it is achieved mutual with PC 101；Upper computer selecting in the present embodiment
PC.
G. by the operating system on PC 101, it is achieved the control to microprocessor 103；
H. internal battery group is arranged at the bottom of main frame 401, remains with external power interface 410 simultaneously；
Microprocessor 103 is made up of temperature microcontroller 104, time microcontroller 107, optical-electric module microcontroller 114,
There is data acquisition and process function；By the operating system of PC 101, microprocessor 103 can be controlled, can enter
Row reaction temperature, response time, data acquisition time, vibrations are initial, the setting of LED light source and control.And letter can be carried out
Number gather and process arrange
Temperature microcontroller 104, time microcontroller 107, optical-electric module microcontroller 114 all use can be according to actual needs
Programming, the microcontroller of data acquisition and processing (DAP).
Reaction thermostatic control system is by temperature microcontroller 104, pwm control circuit 204, heater 105, heater and auxiliary
Device control circuit and temperature sensor 106 is helped to form；Heater 105 selects the heater that metal material makes, internal
Include heater and auxiliary device control circuitry and two groups of metal fins and a desk fan, there is reception temperature micro-control
The instruction of device 104 processed feedback is automatically obtained temp regulating function, temperature controlling range 25-45 DEG C, precision ± 0.1 DEG C；
The microcontroller that this example temperature microcontroller 104 selects model to be P89V51, has 64kB Flash and 1024 bytes
Data RAM, possess ISP and IAP programing function, also there are 3 16 bit timing devices/enumerators, PWM and capture/compare merit
The functions such as PCA, SPI and the UART of energy, meet the accurate control to temperature.
Temperature sensor 106 has collecting temperature and by the signal collected digital data transmission that is converted into temperature microcontroller
The function of 104.
The temperature sensor that in this example, temperature sensor 106 selects model to be DS18B20, this model is by sensor and numeral
Change-over circuit all concentrates in together, temperature-measuring range-55 DEG C-+125 DEG C, and programmable resolution is 9-12 position, minimum distinguishable
Rate is 0.0625 DEG C, reaches device design requirement；
By above combination, after temperature microcontroller 104 passes through pid control computation according to the signal that temperature sensor 106 is fed back
Pwm control circuit 204 is controlled, when temperature is less than the reaction temperature set, then makes heater 105 energising heat up；When
Temperature is higher than the reaction temperature set, then power-off starting fan are lowered the temperature by fin, it is achieved thermostatic control.Temperature
Degree control system realizes closed circuit
Load example reaction room 113 and can carry out 8 sample tests simultaneously, it is also possible to carry out single sample test.
Optical-electric module includes LED light source 109, excites Optical devices 110, transmitting optics 112 and optical detection 111 to form.
LED light source selects MOLD LED LAMP L490 family device, this LED photovoltaic power, illumination and temperature performance full
Foot requirement.
Excite Optical devices by lens 302,306, filter lens 303, dichroscope 304, reflecting mirror 305 form.
Transmitting optics include lens 308,312, filter lens 311, dichroscope 310, reflecting mirror 309 form.
In this example, the fluorescent probe excitation source wavelength of recombinase-mediated isothermal nucleic acid amplification reaction reagent design is
492nm, the wavelength of transmitted light that is stimulated is 518nm, excites Optical devices and transmitting optics by this requirement design.Actual
Can design by fluorescent probe excitation source wavelength and wavelength of transmitted light and excite Optical devices and transmitting optics.
LED light source 109 sends excitation source and pools directional light through lens 302, filters through exciting light filter lens 303 arrowband
After light, remaining 492nm exciting light reflects through dichroscope 304, reflecting mirror 305, then converges to tested through condenser lens 306
On sample 307, sample 307 is stimulated to produce and launches light；
Launch light pool directional light through lens 308, be reflected mirror 309 reflect, dichroscope 310 and filter lens 311 narrow
After band filters, remaining wavelength is the transmitting light of 518nm, converges to photo-detector 111 through condenser lens 312.
Photo-detector 111 receives from the reacted fluorescence signal of sample 307, by photomultiplier tube to receiving
Fluorescence signal is amplified and opto-electronic conversion, becomes the discernible signal of telecommunication, feeds back to optical-electric module microcontroller 114.
The signal detected is sent to the microprocessor 103 of detection device by optical-electric module microcontroller 114.
Shock module 108 is made up of vibrations motor and vibrations motor driver, and vibrations motor driver is by time microcontroller
107 control.
The present embodiment selected to set sample preheating time as 4 minutes, and preheating temperature is 37 DEG C.
Time microcontroller 107, according to the program automatic starting shock motor driver after 4 minutes set, makes vibrations motor
Response sample 307 is carried out vibrations mixing, accelerates response speed；Practical situation according to reaction can also be in the operation of PC
Cancelling vibration function in system, reaction does not carry out vibrations mixing step.
Vibrations motor is connected with response sample room 113, when shaking motor operations, response sample can produce vibrations
Effect.
Response time microcontroller 107 major control response time and vibrations initiate, dwell time, and user can pass through
Response time and vibrations initial time are configured by the operating system on PC, it is possible to according to actual reaction to data
Acquisition time interval is configured.
User can set response time, reaction temperature, data collection interval, shake by the operating system on PC
Dynamic initial time and dwell time.
The present embodiment detects on device and remain with USB communication interface 102, remain two communication interfaces.
This detection device using method comprises the following steps:
A. external power supply or built-in power 115, opening device are connected；
B. communicated by foundation between communication interface 102 with PC 101；
C. response time, reaction temperature, vibrations initial time and dwell time, number are set by PC 101 operating system
According to response parameters such as acquisition time intervals；Response time in the present embodiment is 30 minutes, reaction temperature is 37 DEG C, preheating
Time is 4 minutes, and vibrations initial time is 4 minutes, and during vibrations a length of 30 seconds, data collection interval was 20 seconds.
D. in recombinase-mediated isothermal nucleic acid amplification reaction reagent, the soy bean DNA of variable concentrations is added as template, concentration
Code name respectively L9, L5, L4, L3, L2, L1 and negative controls code name are NTC, in 7 above samples respectively
Add the primer and fluorescent probe designed, put into example reaction room 113, start detection program；
E. detecting one group of data every 20 seconds detection devices, data are sent to microprocessor 114, and PC 101 calls these
Data also analyze and process.
The real-time signal of amplified reaction can be demonstrated by PC and thus draw out instant amplifying curve, as shown in Figure 5.
The foregoing is only the present invention and implement example, not in order to limit the present invention, all the spirit and principles in the present invention it
In, any modification, equivalent substitution and improvement etc. made, should be included within the scope of the present invention.
Claims (17)
1. a recombinase-mediated isothermal nucleic acid amplification reaction real-time detection apparatus, including:
A) microprocessor；
B) reaction thermostatic control system；
C) reative cell of sample is loaded；
D) optical-electric module；
E) response time control module；
F) shock module；
G) at least one communication interface, interacts with host computer.
A kind of recombinase-mediated isothermal nucleic acid amplification reaction real-time detection apparatus the most according to claim 1, it is characterized in that: described microprocessor includes at least temperature microcontroller, response time microcontroller, optical-electric module microcontroller, and has data acquisition and process function.
A kind of recombinase-mediated isothermal nucleic acid amplification reaction real-time detection apparatus the most according to claim 1, it is characterized in that: described microprocessor can be carried out temperature control, response time control, reaction shake control, LED light source control according to user by operating system, outer PC or the instruction of other intelligent apparatus and can carry out signals collecting and process.
A kind of recombinase-mediated isothermal nucleic acid amplification reaction real-time detection apparatus the most according to claim 1, it is characterised in that: described reaction thermostatic control system is made up of temperature microcontroller, PID controller, pwm control circuit, heater and heater auxiliary device control circuitry and temperature sensor.
A kind of recombinase-mediated isothermal nucleic acid amplification reaction real-time detection apparatus the most according to claim 4, it is characterized in that: described heater is the heater of metal or ceramic material, include heater and auxiliary device control circuitry and least one set metal fin and fan, and the instruction receiving temperature microcontroller feedback can be utilized to be automatically obtained temperature regulation, realize temperature controlling range 25-45 DEG C, precision ± 0.1 DEG C.
A kind of recombinase-mediated isothermal nucleic acid amplification reaction real-time detection apparatus the most according to claim 1, it is characterised in that: described temperature sensor has collecting temperature and is converted into the digital data transmission function to temperature microcontroller by signal collected.
A kind of recombinase-mediated isothermal nucleic acid amplification reaction real-time detection apparatus the most according to claim 1, it is characterized in that: described temperature microcontroller can be according to the signal of temperature sensor feedback by being controlled pwm control circuit after PID controller computing, heater energising intensification or power-off starting fan is made to lower the temperature, it is achieved thermostatic control.
A kind of recombinase-mediated isothermal nucleic acid amplification reaction real-time detection apparatus the most according to claim 1, it is characterised in that: described loading example reaction room can carry out 8 sample tests simultaneously, it is also possible to carries out single sample test.
A kind of recombinase-mediated isothermal nucleic acid amplification reaction real-time detection apparatus the most according to claim 1, it is characterised in that: described optical-electric module includes LED light source, excites Optical devices and optical detection system；Optical devices are excited to be made up of one or more groups lens, filter lens, dichroscope, reflecting mirror.
A kind of recombinase-mediated isothermal nucleic acid amplification reaction real-time detection apparatus the most according to claim 9, it is characterised in that: described optical detection system is made up of transmitting optics and photomultiplier tube；Luminous optical device includes one or more groups lens, filter lens, dichroscope, reflecting mirror；Optical detection system receives from the reacted fluorescence signal of sample, is amplified the fluorescence signal received and opto-electronic conversion by photomultiplier tube, becomes the discernible signal of telecommunication, feed back to microprocessor.
11. a kind of recombinase-mediated isothermal nucleic acid amplification reaction real-time detection apparatus according to claim 1, it is characterised in that: described shock module is made up of response time microcontroller, vibrations motor and vibrations motor driver；After sample completes the preheating time that user sets, response time microcontroller is according to the Automatic Program starting shock motor driver set, and makes vibrations motor that response sample to carry out vibrations mixing, accelerates response speed.
12. react real-time detection apparatus according to a kind of recombinase-mediated isothermal nucleic acid amplification described in claim 1 and claim 11, it is characterized in that: described vibrations motor is connected with response sample room, when shaking motor operations, response sample can be produced the effect of vibrations.
13. a kind of recombinase-mediated isothermal nucleic acid amplification reaction real-time detection apparatus according to claim 1, it is characterised in that: described microcontroller major control response time in response time and vibrations initiate, dwell time.
14. a kind of recombinase-mediated isothermal nucleic acid amplifications reaction real-time detection apparatus according to claim 1, it is characterised in that: described communication interface is selected serial ports, USB, bluetooth, Wifi or a combination thereof from including.
15. a kind of recombinase-mediated isothermal nucleic acid amplification reaction real-time detection apparatus according to claim 1, it is characterised in that: described host computer can be PC or other intelligent apparatus.
16. a kind of recombinase-mediated isothermal nucleic acid amplification reaction real-time detection apparatus according to claim 1, it is characterised in that: described detection device using method comprises the following steps,
A. set up between described detection device and host computer by communication interface and communicate；
B. user arranges response time, reaction temperature, vibrations initial time and dwell time, data collection interval by the software system in host computer；Can also create, revise or delete the detection response time of device, reaction temperature, vibrations initial time and dwell time, data collection interval, user can be with self-defined reaction condition and data acquisition conditions.
C. the signal of telecommunication that reaction after starting detects optical detection system is sent to microprocessor, and host computer can call the signal received and utilize software system to be analyzed processing.
17. methods according to claim 16, the signal collected can be shown by host computer and be utilized Software on Drawing to go out instant amplifying curve by user.
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| 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 |
| CN107653301A (en) * | 2017-10-16 | 2018-02-02 | 南京中科拜尔医学技术有限公司 | A kind of hermetic multiple fluorescent PCR detection cartridge |
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| CN108107024A (en) * | 2016-11-25 | 2018-06-01 | 苏州百源基因技术有限公司 | A kind of intelligence PCR instrument |
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| CN109082422A (en) * | 2018-10-23 | 2018-12-25 | 宁波艾捷康宁生物科技有限公司 | A kind of full-automatic PCR nucleic acid extraction detection device |
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| CN109811038A (en) * | 2019-01-17 | 2019-05-28 | 浙江大学 | A kind of bottom illuminated nucleic acid isothermal amplification detection portable instrument |
| CN110066854A (en) * | 2019-05-08 | 2019-07-30 | 中国人民解放军军事科学院军事医学研究院 | A kind of nucleic acid isothermal amplification detection method |
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| CN108107024A (en) * | 2016-11-25 | 2018-06-01 | 苏州百源基因技术有限公司 | A kind of intelligence PCR instrument |
| 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 |
| CN107653301A (en) * | 2017-10-16 | 2018-02-02 | 南京中科拜尔医学技术有限公司 | A kind of hermetic multiple fluorescent PCR detection cartridge |
| CN107937263A (en) * | 2017-11-23 | 2018-04-20 | 北京海普威生物技术有限公司 | Self-check system and method based on gene magnification |
| CN108220123A (en) * | 2018-01-29 | 2018-06-29 | 黄昶荃 | A kind of rapid and handy formula molecular detection devices based on real-time fluorescence quantitative PCR |
| CN110272820A (en) * | 2018-03-13 | 2019-09-24 | 深圳市安鑫宝科技发展有限公司 | Ring mediated isothermal nucleic acid amplification detection device and its application |
| CN112996601A (en) * | 2018-09-14 | 2021-06-18 | 威廉马歇莱思大学 | Apparatus and method for multiplex amplification and detection of DNA using convective heating and label-free microarrays |
| CN109082422A (en) * | 2018-10-23 | 2018-12-25 | 宁波艾捷康宁生物科技有限公司 | A kind of full-automatic PCR nucleic acid extraction detection device |
| CN109337793A (en) * | 2018-10-23 | 2019-02-15 | 宁波艾捷康宁生物科技有限公司 | A kind of full-automatic nucleic acid extraction detection system |
| CN109337793B (en) * | 2018-10-23 | 2022-01-25 | 宁波艾捷康宁生物科技有限公司 | Full-automatic nucleic acid extraction detecting system |
| CN109811038A (en) * | 2019-01-17 | 2019-05-28 | 浙江大学 | A kind of bottom illuminated nucleic acid isothermal amplification detection portable instrument |
| CN110066854A (en) * | 2019-05-08 | 2019-07-30 | 中国人民解放军军事科学院军事医学研究院 | A kind of nucleic acid isothermal amplification detection method |
| CN111443213A (en) * | 2020-04-09 | 2020-07-24 | 星源智（珠海）生物科技有限公司 | Multivariable reaction kinetics real-time detector device and detection method |
| CN114181809A (en) * | 2020-09-14 | 2022-03-15 | 西北农林科技大学 | Nucleic acid amplification product quantitative detection instrument and nucleic acid amplification product quantitative detection method |
| CN113376126A (en) * | 2021-06-08 | 2021-09-10 | 长春长光辰英生物科学仪器有限公司 | Portable loop-mediated isothermal amplification device and operation method thereof |
| CN113376126B (en) * | 2021-06-08 | 2023-12-26 | 长春长光辰英生物科学仪器有限公司 | Portable loop-mediated isothermal amplification device and operation method thereof |
| CN115047925A (en) * | 2022-02-28 | 2022-09-13 | 中国科学院国家空间科学中心 | Passive radiation type constant temperature control system and control method based on PID controller |
| CN115047925B (en) * | 2022-02-28 | 2024-04-30 | 中国科学院国家空间科学中心 | Passive radiation type constant temperature control system and control method based on PID controller |
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