CN102559482B - Gene detection device - Google Patents
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- CN102559482B CN102559482B CN 201110455939 CN201110455939A CN102559482B CN 102559482 B CN102559482 B CN 102559482B CN 201110455939 CN201110455939 CN 201110455939 CN 201110455939 A CN201110455939 A CN 201110455939A CN 102559482 B CN102559482 B CN 102559482B
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Abstract
The invention provides a simple, convenient and quick gene detection device. The gene detection device comprises a gene detector and a controller, wherein the controller controls the gene detection process of the gene detector, acquires and receives scanning data of the gene detector and performs result analysis. The detection process is controlled by a high-precision instrument, so detection speed is greatly increased, detection efficiency is greatly improved, detection cost and manpower resource are effectively saved, and DNA can be detected accurately and exactly at high speed.
Description
Technical field
The present invention relates to the technique of gene detection field, particularly relate to a kind of proofing unit for gene test.
Background technology
1998 the end of the year AAAS classify the electrochemical gene sensor technology one of as big progress in 1998 annual natural science fields ten, it serves to show its meaning in the history of science.It with its can be simultaneously, analyze the ability of thousands of genomic informations quickly and accurately and demonstrated huge power.These are used and mainly comprise genetic expression detection, sudden change detection, genome polymorphism analysis and aspects such as gene library mapping and sequencing by hybridization.
The change of adopting electrochemical gene sensor to detect genetic expression can be saved lot of manpower and material resources and financial resources, former, scientist's change situation that a plurality of genes are observed in a large amount of experiment of repetition of having to, if it almost is inconceivable using conventional methods the change of thousands of genes in the research cell, because must extraction at first extract the nucleic acid of cell, and want abundant to satisfy the needs of Northern hybridization, each probe of mark is hybridized detection more respectively then.Adopt electrochemical gene sensor that thousands of times of ground of workload are reduced, utilize electrochemical gene sensor to detect the function of 6000 genes in the yeast simultaneously, 8600 expression of gene changes among the human fibroblasts have then detected to achievement in the Patrick Brown of Stanford University leader's scientific research group.
Electrochemical gene sensor also can be used for gene sequencing, the present U.S. Human Genome Project is being greatly developed this technology and is being striven for substituting present automatic sequencing, compare with automatic sequencing with existing manual order-checking, the electrochemical gene sensor order-checking can be saved a large amount of reagent and instrument loss.People more successfully comprise that to multiple biology Arabidopis thaliana, yeast and people's genomic expression situation studies in the research that genetic expression detects.Facts have proved that the electrochemical gene sensor technology also can be used for the detection of nucleic acid mutation and the analysis of genome polymorphism, the sudden change that reaches 98% grade with conventional sequencing result consistence detects, to evaluation, mapping and the somatotype of human genome single nucleotide polymorphism, the research of human mitochondria gene group polymorphism etc.
Biosensor is combined with chip technology, by the verified single base mutation that can detect gene of strength of electric field in change probe array zone, thereby by the order of determining overlapping clone the yeast genes group is mapped.
Sequencing by hybridization is another important application of electrochemical gene sensor technology.This sequencing technologies a kind of efficient feasible sequence measurement of can yet be regarded as in theory, but need can derive by the hybridization of a large amount of overlap probes and molecules of interest the sequence of purpose nucleic acid molecule, so need to make a large amount of probes.The electrochemical gene sensor technology can be synthesized and fixing a large amount of nucleic acid molecule with comparalive ease, so its appearance provides the possibility of practice undoubtedly for sequencing by hybridization.
Adopt the electrochemical gene sensor technology, can accelerate the job schedule of the Human Genome Project greatly, for example be used for gene sequencing, genetic expression detects and new genetic marker such as SNP location etc., this is significant to seeking new functional gene, seek new drug target and developing new genomic medicine.Adopt that electrochemical gene sensor can carry out that workload out of the former imagination detects different plant species, different tissues, differently sickly plants, the genetic expression under the different treatment condition changes, thereby know exploitation have different purposes diagnostic kit.New drug must be tested by human safety in the experimental phase, just must observe medicine to the influence of human gene expression, owing to and do not know that medicine works to that a kind of gene, just must the genetic expression in known all or the certain limit all be detected, adopt electrochemical gene sensor can finish this task rapidly and exactly.
But have now by electrochemical sensor and carry out the step that gene test need be more, and for artificial operational requirement than higher, the complex steps time of Jian Ceing is longer simultaneously, simultaneously the situation that a plurality of samples detect is needed lot of manpower and material resources for needs, improved the detection cost, again and, owing to be subjected to the restriction of local and equipment, the very big restriction that obviously has for the processing of various product, particularly for places such as hospital and laboratories, be subjected to the restriction in space, too much equipment can not be installed and hold too much operator, lowered the detection efficiency for sample greatly, than and for needing in the testing process sample is shifted or operates, because a plurality of step separating treatment make sample or data be affected easily, thereby have influenced detection accuracy greatly.
Summary of the invention
The object of the present invention is to provide a kind of easy, be used for the proofing unit of gene test fast.
A kind of proofing unit for gene test, described device comprises:
Gene test device and controller, described controller are controlled the process that described gene test device carries out gene test, gather and receive the scan-data of gene test device, and carry out interpretation of result;
Preferably, described gene test device comprises: electrochemical gene sensor, junctor, the second connection management device and chip heating unit, molecule after described electrochemical gene sensor adopts electrochemical techniques to thing hybridization to be detected carries out mark, realize gene test, the described second connection management device is by the detection signal of alternating voltammetry scanning through mark, and according to the process of the instruction control gene test of described controller, and the temperature of controlling described chip heating unit; Described junctor is used for described electrochemical gene sensor and is connected with the chip heating unit with the second connection management device, and described chip heating unit is used for electrochemical gene sensor is heated.
Preferably, described controller comprises: first management link block, first processing module and second processing module, the described first management link block is connected with described gene test device, receive the scan-data of second connection management device output, and carry out data interaction with the described second connection management device; Described second processing module is carried out interpretation of result to the data of obtaining; Described first processing module is connected with the described first management link block, is used for described gene test device is controlled and managed, and the chip heating unit is controlled, and be used for external communication and connection management.
Preferably, described first processing module comprises: data processing unit and temperature control unit, described data processing unit is used for described gene test device is controlled and managed, and for external communication and connection management, described temperature control unit is used for control chip heating unit, the temperature of regulating electrochemical gene sensor in real time.
Preferably, described proofing unit comprises one or more gene test devices, is used for one or more things to be detected are carried out gene test, and described chip heating unit comprises one or more chip fire ends, is connected with described one or more gene test devices.
Preferably, described temperature control unit comprises: one or more temperature control components and one or more Temperature Feedback element, described one or more temperature control component corresponds respectively to one or more gene test devices, described one or more Temperature Feedback element corresponds respectively to one or more gene test devices, temperature for detection of the gene test device, described one or more temperature control component is followed according to the monitoring temperature of described one or more Temperature Feedback unit to the gene test device, respectively the chip heating unit on one or more gene test devices is controlled in real time.
Preferably, described one or more temperature control component comprises:
Comparing element is used for after the detected temperature simulation digital conversion in one or more Temperature Feedback unit, numerary signal is converted to temperature and target temperature compares;
Controlling elements, result based on the comparison according to parameter prediction temperature variations such as the amplitude of temperature variation, speed, adjusts temperature control work output in real time;
Power element according to the output manipulated variable of controlling elements, is controlled the temperature of described chip heating unit.
Preferably, described junctor comprises:
First coupling end is used for connecting described electrochemical gene sensor;
The environment control end is connected with described chip heating unit, regulates the temperature of control electrochemical gene sensor;
Second electric signal transmission interfaces is connected with the described second connection management device;
Described first coupling end comprises: electrochemical gene sensor fixed bit and first electric signal transmission interfaces that is connected with described electrochemical gene sensor, described electrochemical gene sensor fixed bit is used for fixedly electrochemical gene sensor, and described first electric signal transmission interfaces is used for carrying out data transmission with electrochemical gene sensor;
Described second electric signal transmission interfaces and described first electric signal transmission interfaces interconnect, and are used for transmission of electric signals between the second connection management device and the described electrochemical gene sensor.
Preferably, the second connection management device comprises: digital circuit, modular converter, low-voltage driving module and voltage acquisition reaction circuit;
The work output of the energizing voltage that described Digital Circuit Control low-voltage driving module produces, the voltage acquisition reaction circuit is gathered energizing voltage, and modular converter becomes numerary signal with analog signal conversion, the controller collection comprise voltage acquisition and through the numerary signal that analog signal conversion becomes, detect, control and interpretation of result.
Compared with prior art, the present invention has the following advantages:
Technical scheme of the present invention since intend to adopt each chip independently temperature be controlled to be DNA hybridization highly stable independently environment be provided, and adopt high precision instrument that checkout procedure is controlled, speed and the efficient of check have been improved greatly, and effectively saved cost and the manpower of check, reached at a high speed, accurately and accurately checked DNA.Be used for gene sequencing, compare with automatic sequencing with existing manual order-checking, can save a large amount of reagent and instrument loss.As long as whole detection hybridization and testing process be 30 minutes, and fast and easy, cheapness is compared with order-checking and to have been reduced cost greatly, realizes low-cost detection to the specific gene segment.Can hybridize detection to 24 chips simultaneously, high-throughout examination and the detection of carrying out disease.Whole hybridization and testing process automatization have realized unmanned guard, have alleviated operator's burden, have improved efficient.
Description of drawings
Fig. 1 is the structural representation of proofing unit among the present invention;
Fig. 2 is the structural representation of first processing module among the present invention;
Fig. 3 is the syndeton of junctor among the present invention;
Fig. 4 is temperature controlled synoptic diagram among the present invention.
Embodiment
The present invention is further detailed explanation below in conjunction with the drawings and specific embodiments.Should be appreciated that specific embodiment described herein only is used for explaining the utility model, and be not used in restriction the utility model.
Shown in Fig. 1～4, technical solution of the present invention provides a kind of proofing unit for gene test, and described device comprises:
Gene test device and controller, described controller are controlled the process that described gene test device carries out gene test, gather and receive the scan-data of gene test device, and carry out interpretation of result;
Preferably, described gene test device comprises: electrochemical gene sensor, junctor, the second connection management device, with the chip heating unit, molecule after described electrochemical gene sensor adopts electrochemical techniques to thing hybridization to be detected carries out mark, realize gene test, the described second connection management device is by the alternating voltammetry scanning detection signal through mark, and according to the process of the instruction control gene test of described controller, and the temperature of controlling described chip heating unit; Described junctor is used for described electrochemical gene sensor and is connected with the chip heating unit with the second connection management device.
Wherein, described electrochemical gene sensor comprises: detection zone and electrical signal transmission ends, described detection zone is used for the sample before and after the thing hybridization to be detected is adopted electrochemical techniques, and by the detection signal of alternating voltammetry scanning through mark, be transferred in the described second connection management device via the electrical signal transmission ends, and then make further analyzing and processing, described detection zone can be DNA electrochemical sensor or electronics electrochemical gene sensor etc., can realize the gene detection tool of electrochemical measuring technique.
Described controller comprises: first management link block, first processing module and second processing module;
The described first management link block is connected with described gene test device, receives the scan-data of second connection management device output, and carries out data interaction with the described second connection management module, and described second processing module is carried out interpretation of result to the data of obtaining; Described first processing module is connected with the described first management link block, is used for described gene test device is controlled and managed, and the chip heating unit is controlled, and be used for external communication and connection management.
Preferably, described first processing module comprises: data processing unit and temperature control unit, described data processing unit is used for described gene test device is controlled and managed, and for external communication and connection management, described temperature control unit is used for control chip heating unit, the temperature of regulating electrochemical gene sensor in real time.
Gene test device and controller exist lot of data exchange and numerous control lines, and for reaching high performance hardware connection and being connected configuration flexibly, the first management link block and the two or two connection management device all adopt FPGA as linkage interface among both.FPGA (Field Programmable Gate Array) is field programmable gate array, and it is the product that further develops on the basis of programming devices such as PAL, GAL, EPLD.It occurs as a kind of semi-custom circuit in application specific integrated circuit (ASIC) field, has both solved the deficiency of custom circuit, has overcome the limited shortcoming of original programming device gating circuit number again.The use of FPGA is very flexible, can produce different circuit functions with a slice FPGA by different programming datas.FPGA provides the highest logic density, the abundantest characteristic and the highest performance.Detection module intends adopting the FPGA device of 10-50 ten thousand gate leves, and the tower switchboard adopts the FPGA device of 1,000,000 gate leves.
The FPGA of gene test device connects the generation of this module of control energizing voltage, and energizing voltage comprises choppy sea and sine-wave superimposed, and waveform is produced by DAC by numerary signal; Chip electrode obtains digital quantity by switch arrays with after amplifying through ADC, and ADC is connected control with switch arrays by FPGA.Be connected with FPGA for sensor provides the hybridization environment to get temperature controlling system, realize temperature control and flow rate control.With the manipulated variable of upper module FPGA and the upper processing terminal by the gene test device, the FPGA of controller connects, and realizes the instruction and data communication.
Controller can connect 8 gene test devices, requirement can be simultaneously to 8 sensors scan, data gathering and analyzing and processing.There is a large amount of instruction and data exchanges between controller and gene test device, for realizing data communication at a high speed, adopts FPGA to carry out two kinds of module hardwares and connect.The scan-data of each electrode of the essential analysis of controller simultaneously, the data of each electrode must be carried out data conversion and analysis, count handling of task fast and are finished by the second processing module DSP.DSP obtains data at a high speed by the first management link block (FPGA), and analytical results connects data processing unit (CPU) common data storage area by FPGA equally, and PC directly obtains analytical results from this zone by the CPU notice.
The transaction management of controller is by the CPU(ARM9 treater) finish, CUP need carry out control and management to each module and DSP data analysis are initial.The hardware connection is carried out in being connected by the first management link block (FPGA) of CPU and gene test device, DSP, realizes that CPU carries out control and management to each control object, and the Status Flag of each control object also feeds back to CPU by FPGA.
The data upload PC of controller realizes by network.The embedded ICP/IP protocol stack of CPU software is connected with the network interface card controller on the hardware.System comprises 3 detection towers, and each detection tower is connected with the route of inside, and route is connected with embedded industry PC again, thereby realizes detecting the network connection of tower and PC.Local area network by inside and ICP/IP protocol realize detecting the data exchange of tower and embedded industry PC.
Preferably, described proofing unit comprises one or more gene test devices, is used for one or more things to be detected are carried out gene test, and described chip heating unit comprises one or more chip fire ends, is connected with described one or more gene test devices.
Preferably, described temperature control unit comprises: one or more temperature control components and one or more Temperature Feedback element, described one or more temperature control component corresponds respectively to one or more gene test devices, described one or more Temperature Feedback element corresponds respectively to one or more gene test devices, temperature for detection of the gene test device, described one or more temperature control component is followed according to the monitoring temperature of described one or more Temperature Feedback unit to the gene test device, respectively the chip heating unit on one or more gene test devices is controlled in real time.
Preferably, described one or more temperature control component comprises:
Comparing element is used for after the detected temperature simulation digital conversion in one or more Temperature Feedback unit, numerary signal is converted to temperature and target temperature compares;
Controlling elements, result based on the comparison according to parameter prediction temperature variations such as the amplitude of temperature variation, speed, adjusts temperature control work output in real time;
Power element according to the output manipulated variable of controlling elements, is controlled the temperature of described chip heating unit.
Each sensor of the present invention requires with arriving with inspection, therefore instrument must possess 24 independent detection modules, each module can both be carried out independent temperature control, hybridize the temperature requirements of detecting stages and different test items to satisfy difference, and the control of the temperature of each module must be stablized accurately.In order to reach this requirement, each module adopts independent closed loop thermal control unit, system comprises based on the comparing element of ARM9 treater, based on the controlling elements of PWM output control, based on the power element of heating power pipe, amplifies and the feedback element of ADC based on the heating target (being the chip heating unit) of high thermal conductivity copper base with based on high-precision sensing, as shown in Figure 4;
Test item based on each element of ARM9 Treatment Analysis, and obtain the target temperature of each current module by this project chip agreement and crossing phase, obtain the copper base Current Temperatures by Temperature Feedback element (comprising temperature sensors of high precision, high precision amplification, accurate ADC) simultaneously, after relatively calculating, obtain temperature deviation and manipulated variable, power element control power outputs to heating tube, copper base is carried out temperature control, make it obtain accurately stable temperature.Copper base fully contacts with the chip that inserts detection module, and temperature is transmitted on the chip, for chip hybridization provides suitable temperature environment.
Temperature control algorithm adopts ratio, integration, the differential control based on ARM9, is called for short PID control, claims PID to regulate again.The PID controller algorithm has good stability, reliable operation, advantage easy to adjust, is one of major technique of Industry Control, is used for hybridization temperature control expection of the present invention and can reaches good effect.The structure of controlled plant copper base, parameter and disturbance at random can not be on top of, can not get precise math model, other algorithm of control theory is difficult to adopt, therefore the temperature controlling system structure and parameter must rely on the debugging of experience and development to determine, the thereof using PID control techniques is the most convenient, the PID controller is exactly the error according to system, and proportion of utilization, integration, difference gauge are calculated manipulated variable and controlled.Ratio (P) control is the simplest a kind of control mode, the output of its controller and the proportional relation of error originated from input signal, and system exports and has steady-state error when proportional control only.The output of integration (I) control and error originated from input signal be integrated into proportional relation, temperature controlling system to this instrument, if after entering stable state, there is the steady temperature error, this temperature controlling system has the steady temperature error, in order to eliminate steady-state error, in controller, introduce " integral ", integral depends on the integration of time to error, increase along with the time, integral can increase, even if error is very little like this, integral also can strengthen along with the increase of time, the output increase that it promotes controller further reduces the steady temperature error, up to equalling zero.Therefore, adopt proportional+integral (PI) controller algorithm, can make system's no steady temperature error after entering stable state.The output of differential (D) control and the differential (being the error change rate) of error originated from input signal are proportional, temperature controlling system vibration even unstability can occur in overcoming the regulate process of error, its reason is owing to have big inertia assembly or lag module (lagging behind may appear in copper base heat conduction) is arranged, have the effect that suppresses error, it changes the variation that always lags behind temperature error.For addressing this problem, make the variation " in advance " of the effect that suppresses error, make that when temperature error approaches zero that suppresses error act as zero, what need to increase is " differential term ", the trend that its can predicated error variation.Like this, have stability controller and the algorithm of PID, the control action kou that suppresses temperature error is equalled zero, even be negative value, thereby avoid the serious overshoot of controlled volume.
Preferably, described junctor comprises:
First coupling end is used for connecting described electrochemical gene sensor;
The environment control end is connected with described chip heating unit, regulates the temperature of control electrochemical gene sensor;
Second electric signal transmission interfaces is connected with the described second connection management device;
Described first coupling end comprises: electrochemical gene sensor fixed bit and first electric signal transmission interfaces that is connected with described electrochemical gene sensor, described electrochemical gene sensor fixed bit is used for fixedly electrochemical gene sensor, and described first electric signal transmission interfaces is used for carrying out data transmission with electrochemical gene sensor;
Described second electric signal transmission interfaces and described first electric signal transmission interfaces interconnect, and are used for transmission of electric signals between the second connection management device and the described electrochemical gene sensor.
Preferably, the second connection management device comprises: digital circuit, modular converter, low-voltage driving module and voltage acquisition reaction circuit;
The work output of the energizing voltage that described Digital Circuit Control low-voltage driving module produces, the voltage acquisition reaction circuit is gathered energizing voltage, and modular converter becomes numerary signal with analog signal conversion, the controller collection comprise voltage acquisition and through the numerary signal that analog signal conversion becomes, detect, control and interpretation of result.
Claims (7)
1. proofing unit that is used for gene test is characterized in that described device comprises:
Gene test device and controller, described controller are controlled the process that described gene test device carries out gene test, gather and receive the scan-data of gene test device, and carry out interpretation of result;
Described gene test device comprises: electrochemical gene sensor, junctor, the second connection management device and chip heating unit, molecule after described electrochemical gene sensor adopts electrochemical techniques to thing hybridization to be detected carries out mark, realize gene test, the described second connection management device is by the detection signal of alternating voltammetry scanning through mark, and according to the process of the instruction control gene test of described controller, and the output temperature of controlling described chip heating unit; Described junctor is used for described electrochemical gene sensor and is connected with the chip heating unit with the second connection management device respectively;
Described controller comprises: first management link block, first processing module and second processing module;
The described first management link block is connected with described gene test device, receives the scan-data of second connection management device output, and carries out data interaction with the described second connection management device, and described second processing module is carried out interpretation of result to the data of obtaining; Described first processing module is connected with the described first management link block, is used for described gene test device is controlled and managed, and the chip heating unit is controlled, and be used for external communication and connection management.
2. the proofing unit for gene test according to claim 1, it is characterized in that, first processing module comprises: data processing unit and temperature control unit, described data processing unit is used for described gene test device is controlled and managed, and for external communication and connection management, described temperature control unit is used for control chip heating unit, the temperature of regulating electrochemical gene sensor in real time.
3. the proofing unit for gene test according to claim 2, it is characterized in that, described proofing unit comprises one or more gene test devices, be used for one or more things to be detected are carried out gene test, described chip heating unit comprises one or more chip fire ends, is connected with described one or more gene test devices.
4. the proofing unit for gene test according to claim 3, it is characterized in that, described temperature control unit comprises: one or more temperature control components and one or more Temperature Feedback element, described one or more temperature control component corresponds respectively to one or more gene test devices, described one or more Temperature Feedback element corresponds respectively to one or more gene test devices, temperature for detection of the gene test device, described one or more temperature control component is controlled the chip heating unit on one or more gene test devices respectively in real time according to the monitoring temperature of described one or more Temperature Feedback unit to the gene test device.
5. the proofing unit for gene test according to claim 4 is characterized in that, described one or more temperature control components comprise:
Comparing element is used for after the detected temperature simulation digital conversion in one or more Temperature Feedback unit, numerary signal is converted to temperature and target temperature compares;
Controlling elements, the result according to amplitude, the rate parameter predicted temperature changing conditions of temperature variation, adjusts temperature control work output in real time based on the comparison;
Power element according to the output manipulated variable of controlling elements, is controlled the temperature of described chip heating unit.
6. the proofing unit for gene test according to claim 5 is characterized in that, described junctor comprises:
First coupling end is used for connecting described electrochemical gene sensor;
The environment control end is connected with described chip heating unit, regulates the temperature of control electrochemical gene sensor;
Second electric signal transmission interfaces is connected with the described second connection management device;
Described first coupling end comprises: electrochemical gene sensor fixed bit and first electric signal transmission interfaces that is connected with described electrochemical gene sensor, described electrochemical gene sensor fixed bit is used for fixedly electrochemical gene sensor, and described first electric signal transmission interfaces is used for carrying out data transmission with electrochemical gene sensor;
Described second electric signal transmission interfaces and described first electric signal transmission interfaces interconnect, and are used for transmission of electric signals between the second connection management device and the described electrochemical gene sensor.
7. according to the arbitrary described proofing unit for gene test of claim 1～6, it is characterized in that the second connection management device comprises: digital circuit, modular converter, low-voltage driving module and voltage acquisition reaction circuit;
The work output of the energizing voltage that described Digital Circuit Control low-voltage driving module produces, the voltage acquisition reaction circuit is gathered energizing voltage, and modular converter becomes numerary signal with analog signal conversion, the controller collection comprise voltage acquisition and through the numerary signal that analog signal conversion becomes, detect, control and interpretation of result.
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| CN104914045B (en) * | 2015-05-29 | 2017-09-26 | 重庆大学 | Portable pathogenic bacteria rapid detection system and device |
| CN105331531A (en) * | 2015-10-20 | 2016-02-17 | 上海交通大学 | Portable virus detection device and virus detection method by polymerase chain reaction |
| CN106554920A (en) * | 2016-12-11 | 2017-04-05 | 天津福德信泰生物科技有限公司 | Electrochemical gene detection system |
| CN108441420A (en) * | 2018-05-11 | 2018-08-24 | 南京思达捷信息科技有限公司 | A kind of gene detection system under controller and its method |
| CN112229989A (en) * | 2020-10-19 | 2021-01-15 | 广州吉源生物科技有限公司 | Biological sample identification equipment of GPU (graphics processing Unit) technology |
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