CN106190821A - A kind of hand-held bluetooth PCR instrument being integrated with photoelectric detection system - Google Patents
A kind of hand-held bluetooth PCR instrument being integrated with photoelectric detection system Download PDFInfo
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- CN106190821A CN106190821A CN201610512098.8A CN201610512098A CN106190821A CN 106190821 A CN106190821 A CN 106190821A CN 201610512098 A CN201610512098 A CN 201610512098A CN 106190821 A CN106190821 A CN 106190821A
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L7/00—Heating or cooling apparatus; Heat insulating devices
- B01L7/52—Heating or cooling apparatus; Heat insulating devices with provision for submitting samples to a predetermined sequence of different temperatures, e.g. for treating nucleic acid samples
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2300/00—Additional constructional details
- B01L2300/18—Means for temperature control
- B01L2300/1805—Conductive heating, heat from thermostatted solids is conducted to receptacles, e.g. heating plates, blocks
- B01L2300/1827—Conductive heating, heat from thermostatted solids is conducted to receptacles, e.g. heating plates, blocks using resistive heater
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2300/00—Additional constructional details
- B01L2300/18—Means for temperature control
- B01L2300/1894—Cooling means; Cryo cooling
Abstract
The present invention is a kind of hand-held bluetooth PCR instrument being integrated with photoelectric detection system, belong to molecular biology and medical detecting Instrument field, be made up of metal heat-conducting platform, heating and cooling device, temperature sensor, the control circuit being integrated with blue-tooth device and the photodetector system with PCR reactive tank；PCR reactive tank on metal reaction platform is placed and the fixing PCR reaction tube equipped with reactant, the heating and cooling to reaction platform are controlled by temperature control circuit, temperature sensor feeds back to control circuit to realize precisely regulation after measuring its temperature in real time, final by photoelectric detection system identification performance, the present invention uses simple heating and cooling device and PID to control, while being precisely controlled reaction temperature, ensure that the portability of instrument, course of reaction is monitored by integrated being easy to of blue-tooth device in real time, photoelectric detection system then can determine reaction success or not quickly and easily, avoid some unnecessary resources and temporal waste, effectively reduce experimental cost.
Description
Technical field
The invention belongs to molecular biology and medical detecting Instrument technical field, be integrated with Photoelectric Detection particularly to one
Hand-held bluetooth PCR (Polymerase Chain Reaction, the polymerase chain reaction) instrument of device.
Background technology
Bio information be regulation and control vital movement signal, be constitute organism three big key elements (material, energy,
One of information), organism existence, breeding and metabolism etc. are all played an important role.The wide range that bio information comprises, generally
Refer to DNA, the RNA etc. comprising hereditary information in the narrow sense.These vary with each individual with the sequence of hereditary information, it is also possible to by thing
Reason, the impact of the factor such as chemical, biological and undergo mutation, or make a mistake during self replication, and some duplications, transcribe
Mistake or sudden change can cause the various diseases of human body, such as cancer, thus are analyzed bio information, find out abrupt information in advance
Survey and the important in inhibiting that diagnoses the illness.Analyzing and processing bio information has often required that a certain amount of sample just can carry out gene survey
The work such as sequence, but the gene needed for some Disease-causing gene sequences especially early-stage cancer examination of human body extracting directly gained
The amount of sequence is often the least, therefore needs it is carried out amplification in vitro to reach the magnitude can observed and analyze.Currently, lose
The amplification in vitro passing information sequence is mainly by carrying out polymerase chain reaction in gene-amplificative instrament (PCR instrument)
(Polymerase Chain Reaction) obtains.
The semiconservative replication of DNA is biological evolution and the important channel passed on, and polymerase chain reaction be a kind of for
Amplify the Protocols in Molecular Biology of amplification specific DNA fragments, be regarded as external special semiconservative DNA replication.Continue 1983
United States Biochemical man K.B.Mullis advanced an idea and invented polymerase chain reaction in 1985 year, i.e. simple DNA expands
After increasing method, this technology sustainable development to the third generation, there is heat-resisting, stable archaeal dna polymerase Taq enzyme.PCR reacts first
Utilize the character of degeneration under the DNA high temperature of about 95 DEG C to be in vitro allowed to untwist into strand, then be cooled to about 60 DEG C, make two
Untwist with previous step respectively two strands obtaining of primer are combined by the principle of base pair complementarity, adjust the temperature to 72 the most again
About DEG C (archaeal dna polymerase optimal reactive temperature), make archaeal dna polymerase can hold to the 3 ' directions held along 5 ' with primer for starting point
Complementary strand is synthesized with free base A, T, G, C.Often completing such circulation, the quantity of DNA sequence just doubles, many
After individual circulation, target sequence just can be expanded to considerable magnitude, using the sample as next step analysis of biological information.
Along with life sciences and the development of bioinformatics research, polymerase chain reaction technology and instrument also obtain
More concerns and development, be increasingly becoming part indispensable in life science and experiment.PCR instrument is generally divided into commonly
PCR instrument, grads PCR instrument, In situPCR instrument and real-time fluorescence quantitative PCR instrument four class.Amplified reaction of regular-PCR instrument can only
Set a specified temp, be generally used for specific target gene is expanded, grads PCR instrument then can set a series of not
Same temperature (thermograde), finds optimum reaction temperature in once amplification, more effectively expands；In situPCR
Instrument utilizes the aim sequence fragment that complete cell comes in amplifying cells as a reaction system, and before not destroying cell
Put and detect intracellular amplified production by some particular detection means, it is adaptable to content less target sequence in detection pathological section
Row；Real-time fluorescence quantitative PCR instrument is the regular-PCR instrument with fluorescence signal acquisition system and computer analysis process system, its
Carrying out primer isotope used, fluorescein etc. during PCR amplification labeled, amplification can pass through fluorescence signal acquisition
System Real-time Collection also is transferred in computer analysis process system process to obtain real-time quantized result output.
The consequence all the more shown in life science and medical treatment detection due to PCR instrument and good market
Prospect, many Medical Equipment Plants commercial city has been enter into PCR instrument market, and actively seizes the market share, is wherein no lack of renowned company such as
The ABI U.S. applies biotech firm, ROCHE Roche Diagnistics, MJ, Bole etc., and day, the bright base in Hangzhou, Shanghai are won in domestic Hangzhou
Maple ridge, Xiamen peace Puli etc., driven developing rapidly of global vitro detection market.
Mostly existing PCR instrument, especially domestic equipment, be that build is big towards laboratory and big-and-middle-sized medical institutions, valency
Lattice are high, family and small-sized medical institutions, as community hospital etc. still has no ability to be equipped with.But, due to China's large population base,
Medical resource is the deficientest, and big-and-middle-sized medical institutions can only solve the problem of seeing a doctor of fraction people, and most people cannot enjoy
Carried out disorder in screening or diagnosis by this part medical resource, delay the state of an illness and best occasion for the treatment.Meanwhile, receive along with its people
Entering and growth in the living standard, people pay close attention to all the more healthy, the most growing to the demand of precisely medical treatment.
In sum, PCR instrument etc. is convenient, effective medical diagnostic equipment transfers to basic medical treatment service organization and family
In, PCR instrument effectiveness and market thereof could be maximized, the most effectively solve different medical unit and family's disease detection and diagnosis
The problem of difficulty.
On Vehicles Collected from Market, the PCR instrument volume of main flow is relatively big, heaviness not Portable belt, and operation complexity needs Specialty Experiment personnel to grasp
Make, and the testing time is averagely more than 2 hours.The dyestuff tool of the temporary planless data management system of this kind of PCR instrument and EB dyeing
High carcinogenic, price is had to be also up to several ten thousand RMB.Thus, a kind of volume portable little, light, rapid low one-tenth simple to operate
This, the trend having the PCR instrument device of data management function can be market demand development, replace traditional PCR instrument the most at last and play more
Act on widely.
Summary of the invention
In order to overcome the shortcoming of above-mentioned prior art, it is an object of the invention to provide one and be integrated with photoelectric detection system
Hand-held bluetooth PCR instrument, control heating and cooling device based on PID regulation reaction temperature regulated in real time, coordinate bluetooth
Device realizes the real-time data transmission between computer or cell-phone customer terminal, and utilizes photoelectric detection system Rapid identification experiment knot
Really, its feature includes operation efficient, easy, accurately controls, and be avoided that to react failed product carry out the special of next step again
Property analyze, cause time and cost loss.
To achieve these goals, the technical solution used in the present invention is:
A kind of hand-held bluetooth PCR instrument being integrated with photoelectric detection system, including:
One metal reaction platform 1 placing PCR reaction tube, including lower base and top Re Gai；
One control circuit 2 based on PID/feedback regulation；
One or more heater 3；
One or more heat sink 4；
One or more general temperature sensor 5；
One or more photoelectric detection system 6；
One blue-tooth device 7, for sending the relevant informations such as temperature in real time to receiving device；
It is characterized in that: described control circuit 2 utilizes PID regulation to control heater 3 and heat sink 4 to being placed with
The metal reaction platform 1 of PCR reaction tube carries out heating or lowering the temperature, and utilizes the feedback of temperature sensor 5 to carry out real time temperature to repair
Just, real time temperature information is sent to receiving device so that monitoring in real time by blue-tooth device 7, and utilizes the most afterwards
Photoelectric detection system 6 detects content of predetermined substance in reactant liquor to be changed with Preliminary Identification reaction result.
Described metal reaction platform 1 is made up of the metal of good heat conductivity, includes but not limited to aluminum, aluminium alloy or silver-plated aluminium block
Deng, its as medium heat conductivity than air evenly, than liquid more faster, modes upper and lower, four thorough encirclements can guarantee that again instead
Should pipe variations in temperature evenly.
It is arranged in lower base and the delivery position of top heat lid for placing the reactive tank 101 of PCR reaction tube, it may be assumed that when
When PCR reaction tube is arranged on reactive tank 101, one part is positioned in lower base, and another part is positioned in the heat lid of top.Reaction
Groove 101 is cannelure, and reaction platform top half, i.e. top heat cover and be provided with a PCR reaction tube in each reactive tank 101
The fixing spring 102 of lid, with ensure PCR reaction tube can be fixed on reactive tank 101 closely in and be heated evenly.Metal reaction is put down
Platform 1 main body is cylinder or cuboid or other turntable structure, and outside arranges rotating disk and fixes bayonet socket to 103.Reactive tank 101 is with circle
Radially it is uniformly distributed centered by the axis of a cylinder heart or is evenly distributed in cuboid in a row, to ensure differential responses groove 101
It is heated evenly.
Described control circuit 2 is with MCU as control centre, by writing control program to MCU, to heater 3, cooling dress
Putting 4, temperature sensor 5, photoelectric detection system 6 and blue-tooth device 7 to be scheduling controlling, described MCU passes through USB interface with outer
If or host computer directly communicates；MCU carries out internal storage reading by ISP (In-System Programming) interface
Write and replacement to self, and the data of ISP interface are total by SPI (Serial Peripheral Interface) on sheet
Line is transmitted；On sheet, clock frequency is defined by external crystal oscillator；This MCU can pass through USB with the data transmission of peripheral hardware or host computer
Communication module realizes；Control circuit employs power management module input voltage is changed, for MCU, USB interface and
Other module for power supply.
When the regulation of described control circuit 2 controls reaction temperature, the mode that temperature is arranged includes but not limited to specific amplification
Object sets fixing degeneration, annealing and renaturation temperature, or sets a series of thermograde and find in once amplification
Suitable reaction temperature, in order to follow-up more effectively expand.
Described blue-tooth device 7 is connected with described MCU, receives the data that MCU processed, by communication by data
Real-time Transmission to mobile phone etc. equipped with the equipment of Bluetooth receptions device, it is simple to monitor response situation in real time in device clients.
Described control circuit 2 uses PID control principle to be precisely controlled reaction temperature, when temperature sensor 5 gathers
The set-point desired by dut temperature deviation time, the deviation measuring signal and set-point that PID controls according to feeding back to is carried out
Ratio P, integration I, differential D computing, thus export certain suitable control signal to heater 3 or heat sink 4, promote to survey
Value returns to set-point, reaches the effect automatically controlled.
Described heater 3 include but not limited to heating electric stopband or film heating resistance, described heat sink 4 include but
Being not limited to fan, thermoelectric refrigerator or liquid cooling system, described temperature sensor 5 includes but not limited to temperature-sensitive device or digital temperature
Sensor.
When heater 3 is multiple, should ensure that its heating power is consistent, and be evenly distributed on metal reaction platform 1
Bottom and top, to ensure that reaction platform entirety heats up uniformly.
When heat sink 4 is multiple, should be in parallel to ensure multiple work and power is identical simultaneously.
Described PCR reaction tube is the center arrangement of cincture metal reaction platform 1, heater 3 cloth on metal reaction platform 1
Putting two side position up and down at metal reaction platform 1, position is positioned at upside and the downside of lower base of top heat lid respectively；Described
Heat sink 4 is arranged in metal reaction platform 1 and the outside of heater 3, lay respectively at corresponding heater 3 top or
Below person, described temperature sensor 5 is arranged in the edge of metal reaction platform 1 and the surface location at center.Due to metal reaction
Platform 1 is different with PCR reactant liquor heating rate, therefore still needs to measure in real time and instead after reaction platform temperature reaches period setpoint
Feedback, controls reaction platform temperature and continues for some time to ensure that reactant liquor reaches the temperature consistent with reaction platform.
The judgement of reaction result is to utilize the spectral characteristic of PCR product to enter its concentration by photoelectric detection system 6
Row detection obtains, and described photoelectric detection system 6 carries out Preliminary Identification to product content and (should the most first detect
Once reflection product assay is as benchmark group, this value is generally the least or be zero), to judge whether the reaction result terminated meets
Expection requirement, if product is quantitatively positioned at zone of reasonableness, thinks that reaction is successfully completed, under carrying out product
One step specificity analyses, if product quantity seldom even without; think that reaction is unsuccessful, then need again to carry out to react with
The result.
Described photoelectric detection system 6 includes being arranged in metal reaction platform 1 lower base and the light of Re Gai junction, top
Electricity emitter 601 and photelectric receiver 602, wherein photoemitter 601 is positioned at edge, is specifically arranged in metal reaction platform 1
With between PCR instrument shell, photelectric receiver 602 is positioned at metal reaction platform 1 center, and photoemitter 601 is towards opto-electronic receiver
Device 602, along center position from radial direction to metal reaction platform 1 launch specific wavelength light wave, photelectric receiver 602 towards
Photoemitter 601 direction receives light wave；Light wave, through PCR reaction tube and wherein reactant liquor, judges anti-with the light intensity received
Answer the liquid absorbing state to this light, and then judge the content of product.
Photoemitter 601 and photelectric receiver 602 are fixed in lower base, are positioned at the bottom of metal reaction platform 1
Base and top hot lid intersection, lower edge is concordant with interface, before starting the reaction and terminate after, open top Re Gai, light
The light wave that electricity emitter 601 is launched just can be unobstructed, through PCR reaction tube directive photelectric receiver 602 direction；Photoelectric Detection
Centre-height also can be arranged on lower base and top hot lid intersection by device 6, i.e. photoelectric detection system 6 half is in lower bottom
In Zuo, half is in the heat lid of top, but so arranges, need to be corresponding each between photoemitter 601 and photelectric receiver 602
Reactive tank 101 reserves an optical channel, not only increases manufacturing cost, is also unfavorable for the gentle fall of 1 liter of whole metal reaction platform
The uniformity of temperature.
Described photoemitter 601 includes but not limited to laser diode or light emitting diode, described photelectric receiver 602
Include but not limited to light sensitive diode, photoconductive resistance or photomultiplier tube, the light wave of photoemitter 601 fixed transmission 490nm,
The amount of byproduct of reaction pyrophosphoric acid (PPi) in detection reactant liquor, to judge reaction result.
Hand-held PCR instrument small in volume of the present invention, can gripping with one hand, have portability, relative like product more portable
Band, and simple to operate, test is rapidly, is suitable for basic medical unit and family uses.Control circuit 2 based on PID/feedback regulation
Can realize the accurate regulation of reaction temperature, the integrated of blue-tooth device 7 makes course of reaction to be monitored in real-time, photoelectric detection system
The result of 6 energy anticipation reactions, it is to avoid carry out follow-up detection and specificity analyses in the case of not obtaining expection product,
Reduce detection time and cost to a certain extent.
Compared with prior art, the present invention is integrated with the hand-held bluetooth PCR instrument of photoelectric detection system, by reaction
In real time, it is precisely controlled the application with low cost device and detection means, while realizing high speed, efficient amplification reaction, reaches
Device miniaturization and the human-oriented target of price are arrived, for PCR equipment basic unit with enter family's important in inhibiting.
Accompanying drawing explanation
Fig. 1 is that operation during PCR instrument of the present invention application performs flow chart.
Fig. 2 is the integrally-built generalized section of one of the present invention.
Fig. 3 is the cross section elevational schematic view of a kind of heat lid of the present invention.
Fig. 4 is the sectional overhead schematic view of a kind of PCR instrument main body of the present invention.
Fig. 5 is the circuit diagram of control system of the present invention.
Fig. 6 is the principle schematic of the PID temperature control regulation that the present invention uses.
Fig. 7 is the PID temperature control regulation execution unit schematic diagram that the present invention uses.
Fig. 8 is a kind of heater of using of the present invention and circuit diagram thereof.
Fig. 9 is the structural representation of electro-heat equipment main body in Fig. 8.
Detailed description of the invention
Embodiments of the present invention are described in detail below in conjunction with the accompanying drawings with embodiment.
Operation when a kind of hand-held bluetooth PCR instrument being integrated with photoelectric detection system that the present invention proposes is applied performs stream
Journey as it is shown in figure 1, when reactant liquor prepare complete, put into PCR reaction tube and be placed in reaction platform reactive tank in after, first call
The content of predetermined substance to be detected in reactant liquor is measured by photoelectric detection system, (under normal circumstances should as benchmark group
Value is usually zero or minimum)；Then, covering heat lid, energising starts PCR instrument；In control circuit, MCU is according to each rank of program setting
Section reaction temperature value is transferred heater or heat sink and reaction platform is heated or lowered the temperature, temperature sensor the real time measure
Reaction temperature, this temperature value can be sent to receiving device by blue-tooth device and monitor in real time, need to feed back to MCU simultaneously,
MCU amasss according to the difference of measurement temperature value with set temperature value, application percentage (P) regulation or proportional integral (PI) regulation or ratio
Divide differential (PID) regulation, control heating or temperature-fall period to reach design temperature；When temperature sensor measurement temperature value with set
When constant temperature angle value is consistent, MCU, according to the duration value of program setting, controls heating or heat sink makes the maintenance of reaction platform be somebody's turn to do
The temperature corresponding time；Judge that the current stage of reaction is degeneration, annealing or renaturation, if last stage of reaction completes, then
Think that this is taken turns reaction and terminates, otherwise continue next stage reaction, until epicycle reaction terminates；Each wheel reacts end to reaction wheel
Secondary count, and compare with setting round, if not up to setting round, maintain or update each elementary reaction time and
Temperature (time initial, each phase sets temperature is that fixed value then maintains, and then updates for thermograde), proceeds next round reaction,
If reaching, react stopping；It is again started up photoelectric detection system, launches specific wavelength light wave and specific product content is detected
Tentatively to judge reaction result, if product assay as expected, is thought and is reacted successfully, otherwise, then may need again to test to test
Card result.
Accompanying drawing 2 provides a kind of overall structure figure of the present invention, the i.e. front view of this PCR instrument centre section, whole PCR instrument
Structurally being broadly divided into casing, top heat lid and three parts of sub-body, its middle and upper part heat lid and sub-body contain
All functional parts.Specifically include: a metal reaction platform 1 placing PCR reaction tube；One based on PID/feedback regulation
Control circuit 2；One or more heater 3；One or more heat sink 4；One or more general temperature
Sensor 5；One or more photoelectric detection system 6；And a blue-tooth device 7.
The bosom parts of whole reaction instrument are metal reaction platform 1, are divided into upper and lower two by top heat lid and sub-body
Individual can the part of tight split, wherein reactive tank 101 be divided into similarly upper and lower two can the part of tight split, in reaction
Groove 101 top have PCR reaction lid to fix PCR reaction tube that spring 102 fixes in reactive tank 101 is to ensure that it is anti-with metal
Platform 1 is answered to contact tightly；Reaction platform side is provided with photoemitter 601 (laser diode), solid to reaction Platform center
Surely launch specific wavelength light wave, then have a photelectric receiver 602 (photoconductive resistance) at reaction Platform center, be used for detecting wearing
The intensity of this light wave of PCR reaction tube and reactant liquor injection is tentatively to judge the result that PCR reacts thoroughly.Temperature sensor 5 points
Not being placed in the diverse location of metal reaction platform 1, the temperature data measured applies related algorithm to calculate reaction after passing MCU back
Temperature in pipe, and feeding back to realize being precisely controlled of reaction temperature as the regulation of control system PID.Heater 3, at this
For the folding heating electric stopband uniformly coiled in metal reaction platform 1 upper and lower surface in example, directly connect with metal reaction platform 1
Touching, by MCU control, at degeneration and the renaturation stepwise heating metal reaction platform 1 of reaction, upper and lower surface is symmetrically placed be can ensure that
Heat up evenly.Heat sink 4, is the fan outside metal reaction platform 1 and heater 3, equally by MCU in this example
Controlling, mainly give reaction platform cooling in the annealing steps of reaction, symmetrical above and below being positioned to makes cooling evenly, bottom
Fan can also give simultaneously and control control circuit 2 and lower the temperature.
PCR instrument of the present invention controls heater 3 by control circuit 2 and metal reaction platform 1 is added by heat sink 4
Heat or cooling, measured reaction platform temperature simultaneously in real time, data transmitted in real time by blue-tooth device 7 by temperature sensor 5
To receiving terminal so that user monitors reaction temperature in real time, and feed back to control circuit 2 so that it controls heater 3 and cooling dress
Put 4 pairs of reaction temperatures precisely to regulate.After having reacted, before carrying out next step specificity analyses, right with photoelectric detection system 6
Reaction result carries out preliminary judgement to avoid subsequent step unnecessary time and cost consumption.
Accompanying drawing 3 is the sectional view of a kind of heat lid of the present invention, and hot lid main body is metal reaction platform 1 top, is close to reaction
The heater of platform 3 layers and the fan of more top, in metal reaction platform 1, there is PCR reaction tube the top of reactive tank 101
The fixing spring 102 of lid is in order to be tightly fixed to PCR reaction tube in reactive tank 101.
Accompanying drawing 4 is the sectional view of a kind of PCR instrument main body of the present invention, and metal reaction platform 1 lower base next-door neighbour be placed in
On heater 3 (directly contacting with reaction platform) layer, there is multiple spacing identical in the middle of reaction platform and in reaction platform
The heart is apart from identical reactive tank 101, and the diverse location of reaction platform is also equipped with temperature sensor 5 and reacts platform with real-time measurement
Temperature, heater 3 then has non-close fan and control circuit 2 downwards.During owing to carrying out Photoelectric Detection after having reacted
Need successively the reactant liquor in multiple PCR reaction tubes to be detected, and photoelectric detection system is integrally fixed at cylinder reaction and puts down
The two ends of one radius of platform, therefore need metal reaction platform can rotate around axle center so that each reactive tank position turns to light successively
In the light path of electric detection means (even if each reactive tank place radius overlaps with photoelectric detection system place radius successively), and anti-
Answer platform side equipped with buckle to 103 just can each corresponding inspection positions fix metal platform prevent Photoelectric Detection time
Platform rotates and produces skew, can change to down by hand rotation reaction platform after often having detected a PCR reaction tube and having received data
One PCR reaction tube position and with buckle to 103 fixing reaction platforms.
Accompanying drawing 5 is the circuit diagram of control system of the present invention, and the center of this control system is MCU, and control program is write
Entering MCU and perform, electro-heat equipment, fan, temperature sensor, photoemitter, photelectric receiver, bluetooth module be not respectively by
Being connected and execution corresponding function under it is dispatched with MCU with interface, MCU can be communicated by USB module with peripheral hardware, host computer etc.,
And the data such as temperature also can be sent by bluetooth module in real time, can check equipped with receiving on the equipment of Bluetooth Receiver,
And control circuit employs power management module input voltage is changed, supply for MCU, USB interface and other modules
Electricity.
Accompanying drawing 6 provides the schematic diagram of the PID temperature control regulation that the present invention uses, and accompanying drawing 7 provides the signal of its execution unit
Figure, PID control system is a kind of closed-loop feedback control system, and system initial input is that one or several relatively suitably temperature sets
Holding time of definite value (thermograde) and corresponding temperature, Kp、Ki、KdRepresent that ratio (P), integration (I), differential (D) are made respectively
Coefficient, three jointly acts on and obtains control system transfer function H (s), input quantity transmission function effect under obtain and defeated
Going out controlled quentity controlled variable u, act on object, i.e. react platform, result y is real time temperature measured value, after being fed back to control system,
PID controller recalculates and transfers actuator according to the difference of desired temperature with measured value and adjusts object temperature
Joint, until input value r and output valve y difference are zero.In the present invention, MCU assume responsibility for the work of PID controller, heater and fall
Temperature device to object (reaction platform) to carry out heating or lowering the temperature as the actuator of control system under PID controller control,
Temperature sensor assume responsibility for sending back object (reaction platform) real time temperature the work of MCU as feedback mechanism, and channel radio
The data such as temperature then can be sent to receiving device to realize monitoring in real time by letter module (blue-tooth device) in real time.
Accompanying drawing 8 provides a kind of heater and the circuit diagram thereof that the present invention uses, and electro-heat equipment main body is uniform
The folding heating electric stopband of coiling, as shown in Figure 9, it is evenly distributed on metal reaction platform upper and lower surface (directly contact),
Every resistor is connected in parallel with a capacitor, a termination power, and one end is by complementary Darlington transistor ground connection, and Darlington transistor is opened as power
Closing, its base stage connects MCU pin, controls resistor work under MCU dispatches.
A kind of detailed description of the invention of the present invention is described as follows:
First reactant liquor is quantitatively adding multiple PCR reaction tube, reactant liquor comprise target masterplate DNA, primer, Taq DNA gather
Synthase, 4 kinds of dNTP mixture, amplification buffer, positive divalent magnesium ion and distilled water.PCR reaction tube is put into metal reaction put down
In reactive tank 101 in platform 1, switching on power, first start photoelectric detection system, photoemitter (laser diode) 601 is launched
The light wave of 490nm wavelength, rotational response platform makes light wave sequentially pass through each PCR reaction tube, photelectric receiver (photoconductive resistance)
602 perception projection light intensity substantially to determine the content of pyrophosphoric acid (PPi) in PCR reaction tube, this value will as with reaction result
The benchmark (under normal circumstances, this value is usually zero or minimum) compared；Then, closing heat lid makes PCR reaction lid fix
PCR reaction tube can be tightly fixed in reactive tank 101 by spring 102；According to the control program of write MCU, first start and add hot charging
Put 3 to preheat reaction platform with activating reaction thing, then continue to be heated to about 95 DEG C and maintain certain time, make reaction
Masterplate DNA double chain degeneration in liquid is untwisted becomes two DNA strands separated；In annealing (renaturation) stage, MCU controls to add hot charging
Put 3 quit work, heat sink 4 (i.e. fan) work, reactant liquor is cooled to suitable temperature and maintains a period of time, should
Temperature is generally about 55-60 DEG C, but different according to differential responses thing, at this moment primer and the complementary sequence of masterplate DNA strand
Row combine pairing；In the renaturation stage, heat sink 4 quits work, heater 3 restarts in MCU control, carries out reactant liquor
Being again heated to about 72 DEG C and maintain a period of time, at this moment archaeal dna polymerase reaches most preferably to replicate temperature, thus at archaeal dna polymerase
With 4 kinds of dNTP as reaction raw materials under effect, by base pair complementarity and semiconservative replication principle, close with product on last stage
The DNA double chain of Cheng Xin.During whole, reaction temperature detected by temperature sensor 5 and Real-time Feedback to MCU to carry out PID tune
Joint is also sent out by blue-tooth device, and the actual temp in each stage may be different, all because reactant is different with the persistent period
Given by control program.Three processes of repetitive cycling " degeneration-annealing-extension " repeatedly, can be by required DNA cloning to being easy to detection
Concentration, this period is also given by program, and determine PCR amplification yield, general period is 25-30 time, works as circulation
After number is more than 30, DNA polymerase activity can progressively reach saturated, and the amount of product the most no longer increases with period and increases,
I.e. " plateau " occurs, therefore general period is less than 30.After reaction terminates, opening heat lid, MCU controls photoemitter 601
Launching the light wave of 490nm wavelength, rotational response platform makes light wave sequentially pass through each PCR reaction tube, anti-in detection PCR reaction tube
Answering the concentration of by-product pyrophosphoric acid (PPi), if reacting successfully and fully, enough PPi should be produced, owing to wavelength is by PPi
The light of 490nm has absorbability, and the light intensity through PCR reaction tube directive photelectric receiver 602 can die down, if reacting unsuccessful,
Will not produce or only produce a small amount of PPi, 490nm light not had the strongest trap, then connects through PCR reaction tube directive photoelectricity
The light intensity receiving device 602 is the strongest.According to the characteristic of photoconductive resistance, it can show different resistance under varying strength illumination.By
This can be with preliminary judgement reaction result, if reacting successfully, can carry out the subsequent steps such as electrophoresis, to carry out special to reaction result
Property analyze, if reacting unsuccessful, need not to expend again the time and cost carry out electrophoresis, and may need to re-start PCR reaction
With compound nucleus reaction result.
Claims (10)
1. the hand-held bluetooth PCR instrument being integrated with photoelectric detection system, it is characterised in that including:
One metal reaction platform (1) placing PCR reaction tube, including lower base and top Re Gai；
One control circuit (2) based on PID/feedback regulation；
One or more heater (3)；
One or more heat sink (4)；
One or more general temperature sensor (5)；
One or more photoelectric detection system (6)；
One blue-tooth device (7), for sending the relevant informations such as temperature in real time to receiving device；
It is characterized in that: described control circuit (2) utilizes PID regulation to control heater (3) and heat sink (4) to being placed with
The metal reaction platform (1) of PCR reaction tube carries out heating or lowering the temperature, and utilizes the feedback of temperature sensor (5) to carry out real-time temperature
Degree is revised, and real time temperature information is sent to receiving device so that monitoring in real time by blue-tooth device (7), and before the reaction
After utilize photoelectric detection system (6) detection reactant liquor in content of predetermined substance change with Preliminary Identification reaction result.
It is integrated with the hand-held bluetooth PCR instrument of photoelectric detection system the most according to claim 1, it is characterised in that described gold
Belong to reaction platform (1) to be made up of the metal of good heat conductivity, be arranged in lower bottom for placing the reactive tank (101) of PCR reaction tube
Seat and the delivery position of top heat lid, for cannelure, be provided with a PCR reaction lid and fix bullet in each reactive tank (101)
Spring (102), to ensure that PCR reaction tube can be fixed on closely in reactive tank (101) and be heated evenly.
It is integrated with the hand-held bluetooth PCR instrument of photoelectric detection system the most according to claim 1, it is characterised in that described control
Circuit processed (2) is with MCU as control centre, to heater (3), heat sink (4), temperature sensor (5), photoelectric detection system
(6) and blue-tooth device (7) is scheduling controlling, described MCU is directly communicated with peripheral hardware or host computer by USB interface；
When described control circuit (2) regulation controls reaction temperature, the mode that temperature is arranged includes setting specific amplification object fixing
Degeneration, annealing and renaturation temperature, or set a series of thermograde and find optimal reactive temperature in once amplification, in order to
Follow-up more effectively expand.
It is integrated with the hand-held bluetooth PCR instrument of photoelectric detection system the most according to claim 3, it is characterised in that described indigo plant
Tooth device (7) is connected with described MCU, gives setting equipped with Bluetooth receptions device by communication by real-time data transmission
Standby, it is simple to monitor response situation in real time in device clients.
It is integrated with the hand-held bluetooth PCR instrument of photoelectric detection system the most according to claim 1, it is characterised in that described control
Circuit processed (2) uses PID control principle to be precisely controlled reaction temperature, when the dut temperature that temperature sensor (5) gathers
During set-point desired by deviation, measure signal and the deviation of set-point that PID controls according to feeding back to carry out ratio (P), long-pending
Point (I), differential (D) computing, thus export certain suitable control signal to heater (3) or heat sink (4), promote to survey
Value returns to set-point, reaches the effect automatically controlled.
Be integrated with the hand-held bluetooth PCR instrument of photoelectric detection system the most according to claim 1, it is characterised in that described in add
Thermal (3) is heating electric stopband or film heating resistance, and described heat sink (4) is the cold system of fan, thermoelectric refrigerator or liquid
System, described temperature sensor (5) is temperature-sensitive device or digital temperature sensor.
It is integrated with the hand-held bluetooth PCR instrument of photoelectric detection system the most according to claim 1, it is characterised in that described PCR
Reaction tube is in the upper center arrangement around metal reaction platform (1) of metal reaction platform (1), and heater (3) is arranged in metal
Two side position up and down of reaction platform (1), lays respectively at upside and the downside of lower base of top heat lid；Described heat sink
(4) be arranged in metal reaction platform (1) and the outside of heater (3), lay respectively at corresponding heater (3) top or
Below person, described temperature sensor (5) is arranged in edge and the center of metal reaction platform (1).
It is integrated with the hand-held bluetooth PCR instrument of photoelectric detection system the most according to claim 1, it is characterised in that described light
Electric detection means (6) carries out Preliminary Identification to product content, to judge whether the reaction result terminated meets expection and want
Ask, if product is quantitatively positioned at zone of reasonableness, think that reaction is successfully completed, product is carried out next step special
Specific analysis, if product quantity seldom even without; think reaction unsuccessful, need to again carry out reacting with the result.
It is integrated with the hand-held bluetooth PCR instrument of photoelectric detection system the most according to claim 1, it is characterised in that described light
Electric detection means (6) includes being arranged in the photoemitter at metal reaction platform (1) lower base and top hot lid interface
(601) and photelectric receiver (602), wherein photoemitter (601) is positioned at edge, and photelectric receiver (602) is positioned at center, light
Electricity emitter (601) towards the light wave of photelectric receiver (602) fixed transmission specific wavelength, light wave through PCR reaction tube and
Middle reactant liquor, judges the reactant liquor absorbing state to this light with the light intensity received, and then judges the content of reflection product.
It is integrated with the hand-held bluetooth PCR instrument of photoelectric detection system the most according to claim 9, it is characterised in that described light
Electricity emitter (601) is laser diode or light emitting diode, and described photelectric receiver (602) is light sensitive diode, photosensitive electricity
Resistance or photomultiplier tube, the light wave of photoemitter (601) fixed transmission 490nm, byproduct of reaction Jiao's phosphorus in detection reactant liquor
The amount of acid (PPi), to judge reaction result.
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