CN104865186B - Portable pathogenic bacteria quick determination method - Google Patents
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Abstract
The invention discloses a kind of portable pathogenic bacteria quick determination method, including step 1, magnetic bead is added in test substance, test substance is put into heating tank heated at constant temperature is carried out by temperature control unit, nucleic acid extraction module is put into test substance, the magnetic bead for adsorbing nucleic acid is suctioned out according to the magnetic force area of nucleic acid extraction module；Step 2, test substance is heated by heater, step 3, laser transmitter projects laser, laser is radiated on test substance through laser vias, laser sensor receives the laser passed from laser vias, and after being analyzed by control board, the detection data of test substance are shown in display module.The present invention embodies economy and portability, detects precise and high efficiency.
Description
Technical Field
The invention relates to the field of automatic control, in particular to a portable pathogenic bacteria rapid detection method.
Background
In recent years, biological safety problems caused by frequent food-borne pathogenic microorganism poisoning events, animal and plant pathogen variation or cross-border transmission, safety of transgenic components and the like are more and more prominent. Detection technology and detection equipment in the field of biological safety become one of the hot spots of research. For example, in 2013, the new zealand botulinum milk powder event, Germany cucumber with green cucumber and horse meat adulterated event occur, so that the food safety situation at home and abroad is continuously worsened. Under the background, any technology which is beneficial to the rapid detection of pathogenic microorganisms and gene components thereof has huge application value and market prospect.
The portable gene amplification detection device is developed by firstly utilizing a hand-held magnetic frame to extract nucleic acid DNA in a sample, then adopting a constant-temperature incubator to amplify the nucleic acid, and utilizing a miniature photoelectric detector to analyze and interpret the result. The device developed by the invention not only has special principle, but also is light and portable, and the cost is lower than that of the similar biological gene detection equipment sold in the market. E.g. from DupontA Q7 full-automatic quick detection system for pathogenic microorganism adopts the principle of fluorescence quantitative PCR to detect the microorganism, the BAXQ 7 weight of the system is up to 40 kilograms, and the selling price is up to more than 40 ten thousand RMB. Further, as in BioLumix32 of BioLumix corporation, the number of parallel 32 detection tubes is changed according to the chemical properties of the liquid culture medium of microorganisms, and then the detection is performed by a photosensitive reagent. The weight of the Biolumix32 system was 14 kg. And not to say the sensitivity of the above-mentioned commercialization systemAnd accuracy, which is not suitable for on-site testing of food microorganisms in terms of their weight and selling price. The real-time turbidimeter of Japan Rongand research company and other common PCR instruments can also be applied to the gene rapid detection of food microorganisms, but the nucleic acid extraction component is not integrated, the automation degree is low, the equipment is heavy, and the detection is only suitable for laboratory detection.
Disclosure of Invention
The invention aims to at least solve the technical problems in the prior art, and particularly creatively provides a portable pathogenic bacteria rapid detection method.
In order to achieve the above object, the present invention provides a portable pathogen rapid detection system, which is characterized in that: a power supply unit, a first control chip, a second control chip, a constant temperature control unit, a laser detection unit, a nucleic acid extraction unit, a man-machine interaction module and a communication module,
the power supply unit power output end is connected with the nucleic acid extraction unit power input end, the power supply unit power output end is further connected with the first control chip power input end, the first control chip constant temperature signal transmission end is connected with the constant temperature control unit signal transmission end, the laser detection unit signal transmission end is connected with the first control chip laser signal transmission end, the constant temperature control unit and the laser detection unit power input end are connected with the power supply unit power output end, the first control chip signal output end is connected with the second control chip signal input end through serial port communication, the second control chip signal output end is connected with the human-computer interaction module signal input end, the power supply unit is respectively connected with the second control chip power input end and the human-computer interaction module power input end, the communication module data transmission end is connected with the upper computer data receiving end, and the communication module control chip data transmission end is connected with the first control chip and the second control chip data transmission And (4) an end.
The beneficial effect of above-mentioned scheme does: the control circuit can realize the analysis and judgment of nucleic acid, quickly detect the result and display data in real time.
The invention also discloses a portable pathogenic bacteria rapid detection device, which is characterized by comprising the following components: a nucleic acid extraction module, a heating groove 1, a heating sheet 2, a control circuit board 3, a laser sensor fixing seat 5, a laser sensor 14, a laser emitter 6, a laser emitter lower fixing seat 7, a to-be-detected substance placing hole 22, a to-be-detected substance fixing sheet 23 and a laser via hole 24,
the laser emitter 6 is fixed on the lower laser emitter fixing seat 7, the laser sensor 14 is fixed on the laser sensor fixing seat 5, the laser emitter 6, the heating groove 1 and the laser sensor 14 are sequentially arranged in parallel, the vertical surface of the heating groove 1 is provided with a laser through hole 24, laser emitted by the laser emitter 6 passes through the laser through hole 24 to reach the laser sensor 14,
the top surface of the heating groove 1 is covered with a substance fixing sheet 23 to be measured, a substance placing hole 22 to be measured is formed in the substance fixing sheet 23 to be measured, a groove is formed in the heating groove 1 below the substance placing hole 22 to be measured and used for placing a substance to be measured, and the substance to be measured penetrates through the groove during laser emission;
the heating plate 2 is arranged between the heating groove 1 and the heating groove fixing seat 13;
the nucleic acid extraction module is inserted into a test tube which is filled with a substance to be extracted to extract the nucleic acid of the substance to be detected.
The beneficial effects of the above technical scheme are: by installing the detection device in the detection system, the detection of the substance to be detected is effectively carried out by combining the detection structure of the detection system.
The portable pathogenic bacteria rapid detection device is effective, and the nucleic acid extraction module comprises: handle 16, elastic fixing clip 17, contact rod 19, magnetic force area 20 and pipe sleeve 18,
handle 16 lower extreme installation fixed plate, the fixed plate lower extreme sets up elasticity fixation clamp 17, elasticity fixation clamp 17 is used for fixed pipe box 18, contact rod 19 is fixed in the fixed orifices of fixed plate, the internal thread of fixed orifices cooperatees with contact rod 19's external screw thread, fixes contact rod 19 on the fixed plate, contact rod 19 bottom is magnetized and is produced magnetic field 20, magnetic field 20 is arranged in exploring the recess of heating bath 1 and adsorbs the magnetic bead, handle 16 both sides are connected with the fixed plate by spliced pole 21.
The beneficial effects of the above technical scheme are: the nucleic acid extraction module has reasonable structural design and convenient carrying.
The portable pathogenic bacteria rapid detection device preferably further comprises: the upper fixed seat 15 of the laser emitter is provided with a fixed seat,
the upper fixing seat 15 of the laser emitter is a clamping part formed by folding, the laser emitter 6 is clamped in the middle of the upper fixing seat 15 of the laser emitter, the upper fixing seat 15 of the laser emitter formed by folding is fixed through a through-nail, and the through-nail penetrates through the upper fixing seat 15 of the laser emitter and is nailed into the lower fixing seat 7 of the laser emitter.
The beneficial effects of the above technical scheme are: the fixed seat on the laser emitter is pressed and fixed on the laser emitter, so that the laser emitter is stable and generates no deviation.
The portable pathogenic bacteria rapid detection device preferably further comprises: the heat insulation plate 12 is wrapped around the heating groove 1 and the heating groove fixing seat 13, and light transmission holes are formed in vertical faces of two sides, opposite to the laser through hole 24, of the heat insulation plate 12.
The beneficial effects of the above technical scheme are: the heat insulation plate can prevent a user from being scalded, and the temperature of the substance to be measured in the heating tank is kept constant.
The portable pathogenic bacteria rapid detection device preferably further comprises: and the display module 9 is arranged on the control circuit board 3 and used for displaying data.
The portable pathogenic bacteria rapid detection device preferably further comprises: and the fine adjustment knob 10 is arranged on the periphery below the laser sensor fixing seat 5, and is used for adjusting the height of the laser sensor 14.
The portable pathogenic bacteria rapid detection device, preferably, the control circuit board 3 includes: a first control chip, a second control chip, a constant temperature control unit, a laser detection unit and a power supply,
the laser detection device comprises a first control chip, a second control chip, a laser detection unit, a power supply, a first control chip signal transmission end, a second control chip signal transmission end, a first control chip constant temperature signal transmission end, a constant temperature control unit signal transmission end, a laser detection signal end of the first control chip, and a power supply input end of the first control chip, the second control chip, a display module, the constant temperature control unit and the laser detection unit.
The portable pathogenic bacteria rapid detection device preferably comprises the thermostatic control unit which comprises: a temperature sensor, a heater and a MOS tube,
the utility model discloses a temperature sensor, including heater, power negative pole, MOS pipe source electrode, first control chip PWM pulse modulation end is connected to heater one end connection MOS pipe drain electrode, the heater other end is connected the power anodal, MOS pipe source electrode is connected to the power negative pole, MOS pipe grid connection first control chip PWM pulse modulation end, heater temperature signal output part is connected to temperature sensor temperature signal input part, first control chip temperature signal input part is connected to temperature sensor temperature signal output part.
The beneficial effects of the above technical scheme are: the constant temperature control unit ensures that the substance to be measured in the heating tank is kept constant in temperature, and the effect is remarkable.
The invention also discloses a detection method for the portable pathogenic bacteria rapid detection device, which is characterized by comprising the following steps:
step 1, adding magnetic beads into a substance to be detected, placing the substance to be detected into a heating tank, heating at constant temperature through a constant temperature control unit, placing a nucleic acid extraction module into the substance to be detected, and sucking out the magnetic beads adsorbing nucleic acids according to a magnetic area of the nucleic acid extraction module;
step 2, heating the substance to be detected by a heater, wherein the heating control method comprises the following steps:
the temperature difference and the temperature difference change rate between the real-time temperature and the set value of the ceramic heating plate are used as input quantity of the fuzzy controller, and delta K is obtained through the fuzzy controllerP、ΔKIAnd Δ KDAccording to a self-tuning formula
KP＝KP0+ΔKP
KI＝KI0+ΔKI
KD＝KD0+ΔKD
Calculating a proportional coefficient K after settingPIntegral coefficient KIValue of the differential coefficient KDA value of (1), wherein Δ KPRepresenting the variation of the proportionality coefficient, Δ KIRepresenting the amount of change in the scaling factor; Δ KDRepresenting the variation of the integral coefficient, KP0An initial value representing an integral coefficient; kI0Representing an initial quantity of differential coefficients, KD0An initial value representing a differential coefficient; then K is putP、KIAnd KDThe adjusted value is input into a PID controller, and the differential equation of the PID controller is
The transfer function of the PID controller includes etIs the temperature difference between the real-time temperature and the set value of the ceramic heating plate, dtIs a differential of the time, and is,the differential of the temperature difference between the real-time temperature and the set value of the ceramic heating plate is obtained;
wherein,KPis a proportionality coefficient; t isIIs an integration time constant; t isDIs a differential time constant; d (S) is duty cycle; e (S) is the deviation; u (S) is the output quantity;
in the thermostat control unit, e (k) and ec(k) The temperature difference and the temperature difference change rate between the set value and the actual value are represented, the actual output value u (t) is the duty ratio of the MOS tube, and the controlled object is controlled by controlling the duty ratio;
and 3, the laser emitter emits laser, the laser penetrates through the laser via hole to irradiate on the substance to be detected, the laser sensor receives the laser penetrating out of the laser via hole, and the detection data of the substance to be detected is displayed on the display module after the laser sensor is analyzed through the control circuit board.
In summary, due to the adoption of the technical scheme, the invention has the beneficial effects that:
the invention relates to a portable food-borne pathogenic bacteria rapid detection device, which combines biotechnology and automation technology, adopts the currently international advanced detection method, namely laser detection, judges the negative and positive of a sample by detecting the turbidity of a solution, has relatively simple hardware structure, adopts C language with higher portability as software, can simultaneously detect four groups of samples each time, and improves the detection efficiency. The detection device meets the requirement of outdoor detection, and compared with similar products on the current market, the detection device disclosed by the invention has the following characteristics:
portability
The size of the whole detection device is 255mm 150mm 53mm, compared with the similar detection device in foreign countries, the detection device is small in size and convenient to carry, the detection device in foreign countries must provide a 220V alternating current power supply, and most detection devices are heavy, so that the detection device can only be used in a laboratory and cannot complete outdoor detection work. The power supply used by the invention is a 5V direct current power supply, one lithium battery can meet the working requirement of the lithium battery, and the weight is about 5kg, so that the lithium battery can smoothly complete outdoor detection work.
Economy of use
The superiority of the device can be clearly obtained by comparing the device with the devices of the same type abroad from the economic point of view, and the superiority of the device is provided by Dupont company in the products of the same typeThe selling price of Q7 in China is as high as 40 ten thousand yuan RMB, the cost of the device of the invention is less than one tenth of the cost, and the maintenance of the foreign device is quite troublesome, if the device goes wrong, the device must be sent back to the foreign manufacturer for maintenance, the cycle is longer, the invention uses the domestic technology, the after-sale maintenance is relatively convenient.
Additional aspects and advantages of the invention will be set forth in part in the description which follows and, in part, will be obvious from the description, or may be learned by practice of the invention.
Drawings
The above and/or additional aspects and advantages of the present invention will become apparent and readily appreciated from the following description of the embodiments, taken in conjunction with the accompanying drawings of which:
FIG. 1 is a schematic diagram of a portable pathogen rapid detection system of the present invention;
FIG. 2 is a perspective view of the portable pathogen rapid detection system of the present invention;
FIG. 3 is a schematic diagram of a nucleic acid extraction module of the portable pathogenic bacteria rapid detection system of the present invention;
FIG. 4 is a schematic view of the portable pathogen rapid detection device of the present invention;
FIG. 5 is a circuit diagram of a thermostatic control unit driving circuit of the portable pathogenic bacteria rapid detection device of the present invention;
FIG. 6 is a flow chart of the portable pathogen rapid detection device of the present invention;
FIG. 7 is a circuit diagram of a laser transmitter of the portable pathogen rapid detection device of the present invention;
FIG. 8 is a circuit diagram of a laser sensor of the portable pathogen rapid detection device of the present invention;
FIG. 9 is a flow chart of the method for rapid detection of a portable pathogen according to the present invention;
FIG. 10 is a graph of experimental data for a rapid portable pathogen detection method of the present invention;
FIG. 11 is a graph of experimental data for a rapid portable pathogen detection method of the present invention;
FIG. 12 is a schematic view of a communication module of the portable pathogen rapid detection device according to the present invention;
FIG. 13 is a schematic view of a communication module of the portable pathogen rapid detection device of the present invention.
Detailed Description
Reference will now be made in detail to embodiments of the present invention, examples of which are illustrated in the accompanying drawings, wherein like or similar reference numerals refer to the same or similar elements or elements having the same or similar function throughout. The embodiments described below with reference to the accompanying drawings are illustrative only for the purpose of explaining the present invention, and are not to be construed as limiting the present invention.
In the description of the present invention, it is to be understood that the terms "longitudinal", "lateral", "upper", "lower", "front", "rear", "left", "right", "vertical", "horizontal", "top", "bottom", "inner", "outer", and the like, indicate orientations or positional relationships based on those shown in the drawings, and are used merely for convenience of description and for simplicity of description, and do not indicate or imply that the referenced devices or elements must have a particular orientation, be constructed in a particular orientation, and be operated, and thus, are not to be construed as limiting the present invention.
In the description of the present invention, unless otherwise specified and limited, it is to be noted that the terms "mounted," "connected," and "connected" are to be interpreted broadly, and may be, for example, a mechanical connection or an electrical connection, a communication between two elements, a direct connection, or an indirect connection via an intermediate medium, and specific meanings of the terms may be understood by those skilled in the art according to specific situations.
The portable food-borne pathogenic bacteria rapid detection device takes food-borne pathogenic bacteria as a detection object, detects the change of the turbidity of a sample solution in real time through a laser sensor, transmits a detection signal to a control chip 1, the control chip 1 transmits data to a control chip 2 after calculation processing, the control chip 2 displays the result on a human-computer interaction module, and the working principle of the system is shown in figure 1;
in the figure: 1. a heating groove, 2, a ceramic heating sheet, 3, a control circuit board, 4, a battery, 5, a laser sensor fixing seat, 6, a laser emitter, 7, a laser emitter fixing seat, 8, a fan, 9, a display, 10, a fine adjustment knob, 11, a radiator, 12, a heat insulation board, 13, a heater base, 14, a laser sensor, 15 and a laser emitter fixing seat,
the laser emitter 6 is fixed on the lower laser emitter fixing seat 7, the laser sensor 14 is fixed on the laser sensor fixing seat 5, the laser emitter 6, the heating groove 1 and the laser sensor 14 are sequentially placed in parallel, the laser emitter 6 is fixed on the lower laser emitter fixing seat 7 through a fixing bolt, a first extension end of the control circuit board 3 is connected with the side edge of the lower laser emitter fixing seat 7, the heating groove 1 is placed in parallel with the laser emitter, a laser through hole 24 is formed in the vertical face of the heating groove 1, the laser through hole 24 is used for allowing the laser emitter 6 to emit laser to penetrate through the laser through hole 24 to reach the laser sensor 14, a second extension end of the control circuit board 3 is connected with the side edge of the heating groove base 13, the laser sensor 14 is placed in parallel with the heating groove 1, the laser sensor 14 is fixed on the laser sensor fixing seat 5, and, the emitting end of the laser emitter 6 is in the same straight line with the laser through hole 24 of the vertical surface of the heating groove 1 and the laser sensor 14, the laser emitted by the laser emitter 6 passes through the laser through hole 24 to reach the laser sensor 14,
as shown in fig. 4, the present invention provides a portable pathogen rapid detection system, which is characterized in that the system comprises: a power supply unit, a first control chip, a second control chip, a constant temperature control unit, a laser detection unit, a nucleic acid extraction unit and a man-machine interaction module,
the power supply unit power output end is connected with the nucleic acid extraction unit power input end, the power supply unit power output end is further connected with the first control chip power input end, the first control chip constant temperature signal transmission end is connected with the constant temperature control unit signal transmission end, the laser detection unit signal transmission end is connected with the first control chip laser signal transmission end, the constant temperature control unit and the laser detection unit power input end are connected with the power supply unit power output end, the first control chip signal output end is connected with the second control chip signal input end through serial port communication, the second control chip signal output end is connected with the human-computer interaction module signal input end, and the power supply unit is respectively connected with the second control chip power input end and the human-computer interaction module power input end.
The beneficial effect of above-mentioned scheme does: the control circuit can realize the analysis and judgment of nucleic acid, quickly detect the result and display data in real time.
The human-computer interaction module comprises:
interface design of touch screen
The portable food-borne pathogenic bacteria detection system is a detection system in a touch screen mode. The touch screen must have a touch function in addition to a display function. All instructions of the whole detection process can be realized in a touch mode, and the functions comprise system initialization, parameter setting, starting operation, image processing and the like. Through comparative analysis, a liquid crystal touch screen with the size of 3.5 inches and the resolution of 480 x 320, which is produced by the Delphinai company, is finally selected as the display of the whole device.
Design of press key module
The keyboard can be divided into a matrix keyboard and a non-matrix keyboard according to different connection modes. The matrix keyboard and the non-matrix keyboard have different advantages and disadvantages, and for the matrix keyboard, the matrix keyboard occupies less I/O interface resources of a system, can effectively save the I/O resources of the system, and is suitable for a control circuit with relatively tense I/O interfaces; the non-matrix keyboard occupies a large amount of I/O interface resources, is convenient to use and simple in programming, and is suitable for a control circuit with relatively loose I/O interfaces. For example, a 3 x3 matrix keyboard needs to occupy only 6I/O interfaces, but a non-matrix keyboard with 9 keys also needs to occupy 9I/O interfaces. The controller adopted by the portable food-borne pathogenic bacteria detection system designed by the subject is a 64-pin dsPIC33FJ64GS606 control chip, and an I/O interface is relatively abundant, so that a non-matrix keyboard is selected in the design.
According to the actual requirement of the portable food-borne pathogenic bacteria detection system, the designed non-matrix keyboard is provided with 8 keys, wherein two keys are respectively used for restarting the controller 1 and the controller 2, one key is used for setting the temperature of the constant temperature system, one key is used for setting other parameters of the experiment, one key is used for starting and running the experiment, and the remaining three keys are used as digital input buttons.
Design of SD card function module
In the design, a spiral DNA structure diagram is used as a starting background interface of the whole detection system, but the double-helix DNA structure diagram with the resolution of 480 × 320 and the size of 300K cannot be accommodated because the RAM of the controller is only 64KB, so that an external memory is required to be used, the SD card function is added in the design, and a 2GB SD card is used as the memory of the spiral DNA structure diagram. The SD card has two working modes, an SD mode and an SPI mode.
As shown in fig. 1 and 2, the invention also discloses a portable pathogen rapid detection device, which is characterized by comprising: a nucleic acid extraction module, a heating groove 1, a heating sheet 2, a control circuit board 3, a laser sensor fixing seat 5, a laser sensor 14, a laser emitter 6, a laser emitter lower fixing seat 7, a to-be-detected substance placing hole 22, a to-be-detected substance fixing sheet 23 and a laser via hole 24,
the laser emitter 6 is fixed on the lower laser emitter fixing seat 7, the laser sensor 14 is fixed on the laser sensor fixing seat 5, the laser emitter 6, the heating groove 1 and the laser sensor 14 are sequentially arranged in parallel, the vertical surface of the heating groove 1 is provided with a laser through hole 24, laser emitted by the laser emitter 6 passes through the laser through hole 24 to reach the laser sensor 14,
the top surface of the heating groove 1 is covered with a substance fixing sheet 23 to be measured, a substance placing hole 22 to be measured is formed in the substance fixing sheet 23 to be measured, a groove is formed in the heating groove 1 below the substance placing hole 22 to be measured and used for placing a substance to be measured, and the substance to be measured penetrates through the groove during laser emission;
the heating plate 2 is arranged between the heating groove 1 and the heating groove fixing seat 13;
the nucleic acid extraction module is inserted into a test tube which is filled with a substance to be extracted to extract the nucleic acid of the substance to be detected.
The beneficial effects of the above technical scheme are: by installing the detection device in the detection system, the detection of the substance to be detected is effectively carried out by combining the detection structure of the detection system.
As shown in fig. 3, in the portable pathogen rapid detection device, the nucleic acid extraction module includes: handle 16, elastic fixing clip 17, contact rod 19, magnetic force area 20 and pipe sleeve 18,
handle 16 lower extreme installation fixed plate, the fixed plate lower extreme sets up elasticity fixation clamp 17, elasticity fixation clamp 17 is used for fixed pipe box 18, contact rod 19 is fixed in the fixed orifices of fixed plate, the internal thread of fixed orifices cooperatees with contact rod 19's external screw thread, fixes contact rod 19 on the fixed plate, contact rod 19 bottom is magnetized and is produced magnetic field 20, magnetic field 20 is arranged in exploring the recess of heating bath 1 and adsorbs the magnetic bead, handle 16 both sides are connected with the fixed plate by spliced pole 21.
The beneficial effects of the above technical scheme are: the nucleic acid extraction module has reasonable structural design and convenient carrying.
Nucleic acid extraction module
The nucleic acid extraction module is executed firstly in the whole detection process and is a pretreatment module, and the success or failure of nucleic acid extraction directly influences the final detection result. The nucleic acid is adsorbed by the nucleic acid extraction module mainly by using a magnetic field, so that the purpose of extraction is achieved.
Before the detection is started, a certain amount of magnetic beads are put into a sample to be extracted, then the sample is put into a constant temperature module to be heated, the constant temperature is set to be about 80 ℃, then the nucleic acid extraction device shown in FIG. 3 is put into the sample to be extracted and is driven to move up and down, the biological structure of cells is damaged, DNA is released, the magnetic beads can be combined with the DNA, and under the action of a magnetic field, the magnetic beads are sucked away by a magnetic rod on the nucleic acid extraction device, so that the purpose of extracting the nucleic acid is achieved.
The portable pathogenic bacteria rapid detection device preferably further comprises: the upper fixed seat 15 of the laser emitter is provided with a fixed seat,
the upper fixing seat 15 of the laser emitter is a clamping part formed by folding, the laser emitter 6 is clamped in the middle of the upper fixing seat 15 of the laser emitter, the upper fixing seat 15 of the laser emitter formed by folding is fixed through a through-nail, and the through-nail penetrates through the upper fixing seat 15 of the laser emitter and is nailed into the lower fixing seat 7 of the laser emitter.
The beneficial effects of the above technical scheme are: the fixed seat on the laser emitter is pressed and fixed on the laser emitter, so that the laser emitter is stable and generates no deviation.
The portable pathogenic bacteria rapid detection device preferably further comprises: the heat insulation plate 12 is wrapped around the heating groove 1 and the heating groove fixing seat 13, and light transmission holes are formed in vertical faces of two sides, opposite to the laser through hole 24, of the heat insulation plate 12.
The beneficial effects of the above technical scheme are: the heat insulation plate can prevent a user from being scalded, and the temperature of the substance to be measured in the heating tank is kept constant.
The portable pathogenic bacteria rapid detection device preferably further comprises: and the display module 9 is arranged on the control circuit board 3 and used for displaying data.
The portable pathogenic bacteria rapid detection device preferably further comprises: and the fine adjustment knob 10 is arranged on the periphery below the laser sensor fixing seat 5, and is used for adjusting the height of the laser sensor 14.
The portable pathogenic bacteria rapid detection device, preferably, the control circuit board 3 includes: a first control chip, a second control chip, a constant temperature control unit, a laser detection unit and a power supply,
the laser detection device comprises a first control chip, a second control chip, a laser detection unit, a power supply, a first control chip signal transmission end, a second control chip signal transmission end, a first control chip constant temperature signal transmission end, a constant temperature control unit signal transmission end, a laser detection signal end of the first control chip, and a power supply input end of the first control chip, the second control chip, a display module, the constant temperature control unit and the laser detection unit.
The portable pathogenic bacteria rapid detection device preferably comprises the thermostatic control unit which comprises: a temperature sensor, a heater and a MOS tube,
the utility model discloses a temperature sensor, including heater, power negative pole, MOS pipe source electrode, first control chip PWM pulse modulation end is connected to heater one end connection MOS pipe drain electrode, the heater other end is connected the power anodal, MOS pipe source electrode is connected to the power negative pole, MOS pipe grid connection first control chip PWM pulse modulation end, heater temperature signal output part is connected to temperature sensor temperature signal input part, first control chip temperature signal input part is connected to temperature sensor temperature signal output part.
The beneficial effects of the above technical scheme are: the constant temperature control unit ensures that the substance to be measured in the heating tank is kept constant in temperature, and the effect is remarkable.
Constant temperature control module
The constant temperature control module is an important part of the device, and the food-borne pathogenic bacteria need proper environmental temperature in the rapid value-added process, so that a constant temperature module needs to be designed to meet the requirement. The main circuit schematic diagram of the thermostatic module is shown in fig. 5:
the controller selects dsPIC33FJ64GS606 in Microchip series, and an independent PWM generator and a 10-bit A/D sampler are arranged in the controller, so that an external A/D chip is avoided, and the whole circuit structure is simplified; the heater adopts a 5V ceramic heating plate, the heating groove is formed by processing an aluminum block, and the temperature sensor adopts DS18B20 of Dallas semiconductor company. The constant temperature setting value of the constant temperature module is generally over 60 ℃, and in order to avoid the great influence of the constant temperature module on other modules, the heat insulation plates are respectively added on the periphery of the constant temperature module, so that the influence on other modules can be reduced, and the performance of the constant temperature module can be improved. According to the parameter characteristics of the circuit, the MOSFET driving circuit is selected as follows:
the isolation chip adopts a double-channel optical coupling isolation HCPL _2631, the drive chip adopts a double-channel MAX17600, the whole detection device is designed into four-channel sampling, a multi-channel drive circuit is needed, and the double-channel isolation and drive chip is favorable for simplifying the circuit.
As shown in fig. 6, the controller controls the output power of the ceramic heating plate through the duty ratio of the MOSFET, thereby achieving the purpose of controlling the temperature. The larger the duty ratio is, the larger the output power of the ceramic heating sheet is, when heating is carried out with a certain duty ratio, the temperature sensor transmits the temperature of the heating groove to the controller in real time, the value of the duty ratio is changed after the controller calculates and judges, the output power of the heating sheet is changed through continuously correcting the duty ratio, and then the temperature of the heating groove is constant. The device adopts fuzzy PID control, the fuzzy PID controller can not depend on the mathematical model of the controlled object, and is suitable for the controlled object of the device, and the fuzzy PID controller is easier to realize.
The laser detection technology is one of the most advanced technologies in the present generation, and has been applied in more and more fields, and compared with the traditional chemical detection method, the detection precision is higher, and the use is more convenient. The device adopts laser detection, and the detection result is estimated by detecting the change of the turbidity of the solution through the laser. In order to improve the working efficiency of detection, the device designs four-path simultaneous detection function, and consists of four-path laser emission circuits and four-path laser detection circuits, wherein each path of laser emission circuit is shown in figure 7:
because laser emitter's temperature can obviously rise when working for a long time to lead to laser emitter's output to diminish, and then make the testing result great deviation appear, in order to solve this problem, designed a temperature control system, be used for guaranteeing that laser emitter's temperature is stable in certain extent. Fix laser emitter on metal base, metal base puts in refrigeration piece top, and the working method of refrigeration piece is controlled by outside control circuit, and when laser emitter's high temperature, the refrigeration piece begins work, and the temperature that makes the transmitter through heat-conduction effect reduces, and when the temperature of transmitter is less than the setting value, the refrigeration piece stop work makes laser emitter's temperature stabilization in certain extent through this kind of mode to improve the precision of testing result.
When laser emitted by the laser emitter penetrates through the sample solution and then irradiates on the laser receiver, the laser receiver converts an optical signal into an electric signal, the stronger the light intensity of the received laser is, the larger the generated photocurrent is, the photocurrent is converted into voltage through the resistor RL, and the sampling voltage is measured by the A/D sampler. If the turbidity of the solution changes, the light intensity of the solution passing through the solution changes, the photocurrent generated by the receiver also changes, and the sampling voltage also changes, so that the detection result can be judged by adopting the change of the voltage. R1 and C1 constitute a filter circuit, the power supply adopts 3.3V direct current power supply, as shown in figure 8, FDS100 is selected as the laser sensor, and a 10-bit A/D module in the controller is selected as the A/D sampler. The sampling time can be adjusted according to the working time of the whole detection device.
Theoretical analysis shows that when the sample to be detected is negative, the sample to be detected does not contain food-borne pathogenic bacteria, the quantity of the food-borne pathogenic bacteria in the sample cannot be amplified after the sample is heated at constant temperature, and the solution or turbidity basically keeps unchanged in the whole detection process, so that the obtained sampling voltage value also basically keeps unchanged in the whole detection process, and the obtained sampling curve is basically a parallel straight line and is lower than a threshold line. The threshold line is a standard line derived by experts in the biological field and used for judging whether the sample is positive or negative, when a sampling curve has a sampling value higher than the standard line, the detected sample is positive, and when the sampling curve is always lower than the standard line, the detected sample is negative.
Because the constant temperature control module, the nucleic acid extraction module and the laser detection module are all arranged in the same device, the working temperature of each component is fully considered to dissipate heat. The temperature of the constant temperature module is generally higher than 60 ℃, while the temperature of the nucleic acid extraction module is higher when the nucleic acid extraction module works, even exceeds 80 ℃, the high temperature of the two modules can affect other modules, especially the temperature stability of the laser emitter has a fatal influence, if the laser emitter can not work stably, the detection circuit can change along with the change of the emitter power, and the detection accuracy can be greatly affected. Four heat insulation plates are arranged on the periphery of the constant temperature module and the periphery of the nucleic acid extraction module to slow down the heat exchange effect of the constant temperature module and the nucleic acid extraction module with other modules, an aluminum radiator with a larger volume is arranged at the lower end of the laser emitter, and 4 paths of fans are arranged beside the radiator to enhance air circulation and enhance the heat dissipation capability.
The portable device can be realized without the support of a human-computer interaction software technology, and the software meets the requirements of functional integrity, practicability, easy operability, attractiveness and the like. The software system adopts the design concept of modularization and integration. The instrument parameter setting module is used for setting the working time of the detection device, the constant temperature value of the constant temperature module and the on/off of the working state of the four-path laser detection; the A/D sampling module is used for sampling the detection result. And transmitting the result to the controller; the PID control module is used for constant temperature regulation to stabilize the temperature at a set value; the serial communication is used for data transmission between the two controllers, data of the controller 1 needs to be transmitted to the controller 2 first when the data needs to be displayed on the display, and the data needing to be displayed is displayed on the human-computer interaction module through the controller 2; and the detection result display module is used for displaying the detected experimental data and the temperature value.
Communication module
The communication module of the portable food-borne pathogenic bacteria detection system is mainly divided into two parts, wherein the first part is communication between the portable food-borne pathogenic bacteria detection system and an upper computer; the second part is the mutual communication between the two main control chips, and the structure of the communication module is shown in figure 12;
the communication module mainly comprises a communication module between the portable food-borne pathogenic bacteria detection system and the upper computer and a communication module between the controllers, and each communication module comprises a hardware design part and a software design part.
The communication types are various, and for convenience of connection, a USB interface is adopted, and USB communication is converted into serial communication through a chip CH340, so that the use of a complex USB communication protocol is avoided, and a specific circuit diagram is shown in FIG. 13.
Control algorithm
As shown in fig. 9, when designing the constant temperature module, a control method needs to be applied to the controlled object, and after considering the performance parameters of the constant temperature module, the fuzzy PID control is selected, because the mathematical model of the controlled object of the device is uncertain, and can satisfy each performance index of the constant temperature module, and the fuzzy PID is easy to implement.
The proportional-integral controller is the most widely used one at present, and is mainly used for liquid level, pressure, flow and other control systems in industrial production. The integration effect is introduced to eliminate residual difference, make up the defect of pure proportion control and obtain better control quality. However, the introduction of the integration effect deteriorates the system stability. For control systems with large inertial lags, use is to be avoided as much as possible.
The invention has two fuzzy self-tuning PID controllers in total, wherein one is a constant temperature system of a laser transmitter, the other is a constant temperature system of a heating tank, and actuators of the two constant temperature systems are the same and are the duty ratio of an MOSFET (metal-oxide-semiconductor field effect transistor); the controlled objects are different, the controlled object of the laser emitter constant temperature system is a refrigerating sheet, and the controlled object of the heating groove constant temperature system is a ceramic heating sheet.
The controlled object in the constant temperature system is a ceramic heating plate or a refrigerating plate, therefore, the input quantity of the fuzzy PID controller of the constant temperature system is the real-time temperature of the ceramic heating plate or the deviation value e between the real-time temperature of the refrigerating plate and the corresponding set temperature1(k) Amount of change e from deviationc(k) The output quantity is three parameters of the PID controller, namely delta KP、ΔKIAnd Δ KD。
E is to be1、ec、ΔKP、ΔKIAnd Δ KDThe fuzzy sets of (a) are set as: { NB, NM, NS, ZE, PS, PM, PB }, wherein NB, NM, NS, ZE, PS, PM, PB respectively represent negative big, negative middle, negative small, zero, positive small, middle, positive big; the fuzzy set domain is { -3, -2, -1, 0, 1, 2, 3 }.
Input quantity e1、ecAnd an output Δ KP、ΔKIAnd Δ KDThe actual value range of (a) is the basic universe of discourse of the system. Quantities in the fundamental domain are continuously valued analog quantities. Let deviation e1Has a basic discourse field of [ -x, x]If there is a deviation e1The domain of fuzzy set argument of (a) is { -n, - (n-1), …, n-1, n }, then the precise quantity e is defined1Has a fuzzy quantization factor of KeComprises the following steps:
similarly, the deviation variance e can be definedcQuantization factor of
2) Determination of membership functions
Common membership functions are triangular, trapezoidal and gaussian. Since the triangular membership function is the simplest, the triangular function is selected as the membership function of the deviation and the deviation variation, as shown in fig. 10, and the triangular function is selected as the membership function of the correction quantity of the three parameters of the PID controller, as shown in fig. 11.
Setting of controller rules
Three control parameters K of the PID controller are comprehensively considered from the aspects of stability, response speed, overshoot, steady-state precision and the like of the systemP、KI、KDThe fuzzy control rule can be obtained as follows:
first, in PID control, the response speed of the system is represented by KPAnd (6) determining. In order to obtain good performance of the system, the larger K should be selected appropriately when in the initial stage of regulationPValues to increase system response speed; when in the middle of regulation, KPA smaller value should be selected so as to ensure that the system has a smaller overshoot while ensuring a certain response speed; at the end of the regulation, KPA larger value should be chosen in order to reduce the steady state error of the system and improve the control accuracy.
Second, the integration action can eliminate the steady state error of the system. When in the initial stage of the regulation, a phenomenon of integral saturation may occur due to some factors such as saturation nonlinearity, resulting in a large overshoot amount. Therefore, it is made ofAvoiding integral saturation, and weakening the integral part in the initial stage of regulationIMay even be zero; in order to avoid influencing the system stability when in the middle of regulation, KIA moderate value should be selected; when the adjustment is at the end stage, the integral action needs to be properly increased so as to reduce the steady-state error of the system and improve the control precision.
Thirdly, the differential loop can predict the trend of error change in energy saving mode, so that the control action of the suppression error is equal to zero in advance, even is a negative value, thereby avoiding the serious overshoot of the controlled quantity and improving the dynamic characteristic of the system in the adjusting process. According to practical experience, combined with theoretical analysis, in the initial stage of regulation, the differential action should be increased, i.e. a greater K is takenDA value such that there is little or no overshoot; in the middle of regulation, KDSelecting a value with a proper size; at the end of the adjustment, K should be decreasedDIn order to impair the braking action of the controlled process.
From the above analysis, Δ K is derived from the "IF A AND B THEN C" fuzzy statementP、ΔKIAnd Δ KDThe fuzzy control rules of (1) are shown in table 1, table 2 and table 3, respectively.
TABLE 1. DELTA.KPFuzzy control rule table
TABLE 2. DELTA.KIFuzzy control rule table
TABLE 3. DELTA.KDFuzzy control rule table
4) Defuzzification
The process of extracting the accurate output from the output fuzzy set is called defuzzification, which is also called sharpening, defuzzification and defuzzification. The common methods for defuzzification of the fuzzy quantity include a maximum membership method, a median method and a gravity center method. The centroid method includes and utilizes all information of the fuzzy set, and has a certain emphasis according to different membership degrees, so that the centroid method is widely applied.
The gravity center method is suitable for realizing defuzzification, because the output value of the inference of the gravity center method changes when the input signal has small change. The centroid method is essentially a weighted average method and can be expressed by the following equation:
wherein u is*Is a precise amount, xiIs the value of the corresponding element in the output argument domain,denotes xiDegree of membership.
Defuzzification to obtain u*It also needs to be converted into an exact quantity u in the fundamental domain. Let the basic argument of the output u be [ -y, y]Where the ambiguity field is { -n, - (n-1), …, n-1, n }, and the scaling factor K is defined as y/n, then this is available
5) P, I, D self-tuning
Deriving Δ K from a fuzzy controllerP、ΔKIAnd Δ KDAfter the output value is obtained, P, I, D parameters are corrected by utilizing the notations 5.1, 5.2 and 5.3 to obtain a new P, I, D parameterThe value is obtained.
KP＝KP0+ΔKP(5.1)
KI＝KI0+ΔKI(5.2)
KD＝KD0+ΔKD(5.3)
The controller can only process digital signals, PID control is required to be discretized, P, I, D is used as an output value, the duty ratio of the MOSFET is used as an output value, the output power of the refrigerating sheet or the output power of the ceramic heating sheet is controlled through the duty ratio, and the purpose of constant temperature is finally achieved.
In the description herein, references to the description of the term "one embodiment," "some embodiments," "an example," "a specific example," or "some examples," etc., mean that a particular feature, structure, material, or characteristic described in connection with the embodiment or example is included in at least one embodiment or example of the invention. In this specification, the schematic representations of the terms used above do not necessarily refer to the same embodiment or example. Furthermore, the particular features, structures, materials, or characteristics described may be combined in any suitable manner in any one or more embodiments or examples.
While embodiments of the invention have been shown and described, it will be understood by those of ordinary skill in the art that: various changes, modifications, substitutions and alterations can be made to the embodiments without departing from the principles and spirit of the invention, the scope of which is defined by the claims and their equivalents.
Claims (2)
1. A portable pathogenic bacteria rapid detection method is characterized by comprising the following steps:
step 1, adding magnetic beads into a substance to be detected, placing the substance to be detected into a heating tank, heating at constant temperature through a constant temperature control unit, placing a nucleic acid extraction module into the substance to be detected, and sucking out the magnetic beads adsorbing nucleic acids according to a magnetic area of the nucleic acid extraction module;
step 2, heating the substance to be detected by a heater, wherein the heating control method comprises the following steps:
in the fuzzy PID controller, the performance of the system is determined by fuzzy control and PID control together, the fuzzy control can set P, I, D parameter values, and the response speed of the system is determined by the integral action; the integral action eliminates the steady-state error of the system; the differential loop can predict the trend of error change in an energy-saving manner, and the control action of the suppression error is equal to zero or is a negative value in advance;
in a constant temperature system, a temperature difference e (k) and a temperature difference change rate e between a set value and an actual value are definedc(k) The actual output value u (t) is the duty ratio of the MOS tube, and the ceramic heating sheet is controlled by controlling the duty ratio to achieve the purpose of heating the substance to be detected;
and 3, the laser emitter emits laser, the laser penetrates through the laser via hole to irradiate on the substance to be detected, the laser sensor receives the laser penetrating out of the laser via hole, and the detection data of the substance to be detected is displayed on the display module after the laser sensor is analyzed through the control circuit board.
2. The method for rapidly detecting a portable pathogen according to claim 1 wherein the step 2 comprises:
the temperature difference and the temperature difference change rate between the real-time temperature and the set value of the ceramic heating plate are used as input quantity of the fuzzy controller, and delta K is obtained through the fuzzy controllerP、ΔKIAnd Δ KDAccording to a self-tuning formula
KP＝KP0+ΔKP
KI＝KI0+ΔKI
KD＝KD0+ΔKD
Calculating a proportional coefficient K after settingPIntegral coefficient KIValue of the differential coefficient KDA value of (1), wherein Δ KPRepresenting the variation of the proportionality coefficient, Δ KIRepresenting the amount of change in the scaling factor; Δ KDRepresenting the variation of the integral coefficient, KP0An initial value representing an integral coefficient; kI0Representing an initial quantity of differential coefficients, KD0An initial value representing a differential coefficient; then K is putP、KIAnd KDThe adjusted value is input into a PID controller, and the differential of the PID controllerThe equation is
<mrow> <mi>u</mi> <mrow> <mo>(</mo> <mi>t</mi> <mo>)</mo> </mrow> <mo>=</mo> <msub> <mi>K</mi> <mi>P</mi> </msub> <mo>&lsqb;</mo> <msub> <mi>e</mi> <mi>t</mi> </msub> <mo>+</mo> <mfrac> <mn>1</mn> <msub> <mi>T</mi> <mi>I</mi> </msub> </mfrac> <munderover> <mo>&Integral;</mo> <mn>0</mn> <mi>t</mi> </munderover> <msub> <mi>e</mi> <mi>t</mi> </msub> <msub> <mi>d</mi> <mi>t</mi> </msub> <mo>+</mo> <msub> <mi>T</mi> <mi>D</mi> </msub> <mfrac> <msub> <mi>d</mi> <msub> <mi>e</mi> <mi>t</mi> </msub> </msub> <msub> <mi>d</mi> <mi>t</mi> </msub> </mfrac> <mo>&rsqb;</mo> </mrow>
The transfer function of the PID controller includes etIs the temperature difference between the real-time temperature and the set value of the ceramic heating plate, dtIs a differential of the time, and is,the differential of the temperature difference between the real-time temperature and the set value of the ceramic heating plate is obtained;
wherein, KPIs a proportionality coefficient; t isIIs an integration time constant; t isDIs a differential time constant; d (S) is duty cycle; e (S) is the deviation; u (S) is the output quantity;
in the thermostat control unit, e (k) and ec(k) The temperature difference and the temperature difference change rate between the set value and the actual value are represented, the actual output value u (t) is the duty ratio of the MOS tube, and the controlled object is controlled by controlling the duty ratio.
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| CN113189899A (en) * | 2021-03-23 | 2021-07-30 | 广州万孚倍特生物技术有限公司 | Hardware control device and system of medical instrument |
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