CN113485478A - Temperature control method, device and system of PCR amplification instrument, computer equipment and computer readable storage medium - Google Patents
Temperature control method, device and system of PCR amplification instrument, computer equipment and computer readable storage medium Download PDFInfo
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Abstract
The invention relates to the technical field of automatic control, and discloses a temperature control method, a device, a system, computer equipment and a computer readable storage medium of a PCR amplification instrument, namely, a comparison result of a real-time target temperature difference and a preset temperature difference threshold value is used as a decision basis for switching between a fuzzy control mode and a fuzzy PI D control mode, the fuzzy control mode can be applied when the target temperature difference is larger, so as to ensure that the temperature is increased/reduced at the fastest speed, and the fuzzy PI D control mode is applied when the target temperature difference is smaller, so as to ensure low steady-state error and low overshoot, realize the aim of accurately maintaining the temperature, further ensure that a temperature control system of the PCR amplification instrument has a multi-mode temperature control mode, can ensure that the system performance achieves the unification of high temperature increase and decrease speed, low overshoot, small oscillation and high precision to the utmost extent, and improve the amplification efficiency of DNA fragments, the time required by the reaction is shortened, the PCR detection result can be rapidly obtained, and the practical application and popularization are facilitated.
Description
Technical Field
The invention belongs to the technical field of automatic control, and particularly relates to a temperature control method, a temperature control device, a temperature control system, computer equipment and a computer readable storage medium of a PCR amplification instrument.
Background
Polymerase Chain Reaction (PCR) is a method of using a DNA (deoxyribose Nucleic Acid) as a template, and amplifying the DNA to a sufficient amount in the presence of DNA Polymerase and nucleotide substrates for structural and functional analysis. The PCR detection method has very important significance in clinically and rapidly diagnosing bacterial infectious diseases and the like.
The PCR amplification instrument is a common instrument for realizing the rapid amplification of specific DNA fragments in vitro, and the basic process for carrying out PCR reaction comprises the following three steps: (1) DNA denaturation (with a working temperature of 94 ℃) is carried out, even if the double-stranded DNA template is broken by hydrogen bonds under the action of heat, single-stranded DNA is formed; (2) annealing (the working temperature is 55 ℃), and even if the system temperature is reduced, the primer is combined with the DNA template to form a local double strand; (3) extension polymerization (the working temperature is 72 ℃), i.e., under the action of Taq enzyme (DNA polymerase with thermal stability separated from Thermus Aquaticus) and with dNTP (deoxyribose-nucleotide triphosphate) as raw material, a DNA chain complementary to the template is synthesized by extension from 5 end to 3 end of the primer. The PCR amplification instrument controls the sample to reach different working temperatures, and performs denaturation, annealing, extension polymerization and the like on the amplified DNA fragments so as to achieve the aim of amplifying the quantity of the DNA fragments in multiples. It can be seen that in the PCR amplification apparatus, the accuracy of temperature control, especially the time control of each temperature value, directly affects the efficiency of DNA fragment amplification.
However, the temperature control method of the existing PCR amplification apparatus generally only adopts a fuzzy PID (proportional, Integral, and Differential english abbreviation) control method to control the temperature, so that the control time is longer in the temperature rise and fall interval with large temperature change, and the efficiency of the whole PCR amplification reaction process is not high, and needs to be optimized.
Disclosure of Invention
In order to solve the problem that the temperature control method of the existing PCR amplification instrument influences the amplification efficiency of DNA fragments due to the fact that the control time of a temperature rising and falling interval is long, the invention aims to provide a novel temperature control method, a device, a system, computer equipment and a computer readable storage medium which are applied to the PCR amplification instrument, so that the performance of a temperature control system can be unified to the greatest extent with high temperature rising and falling speed, low overshoot, small oscillation and high precision, the amplification efficiency of the DNA fragments is improved, the time required by reaction is shortened, the PCR detection result can be obtained quickly, and the practical application and popularization are facilitated.
In a first aspect, the present invention provides a method for controlling temperature of a PCR amplifier, which is performed by a controller in the PCR amplifier, and includes:
acquiring a real-time temperature value acquired by a temperature sensor in the PCR amplification instrument at a current sampling time node;
calculating to obtain a temperature difference value between the real-time temperature value and a target temperature value, wherein the target temperature value corresponds to the current sampling time node;
judging whether the absolute value of the temperature difference value is larger than a preset temperature difference threshold value, wherein the preset temperature difference threshold value is between 2 and 3 ℃;
if yes, controlling a first fuzzy controller in the PCR amplification instrument to be in a working state, controlling a second fuzzy controller and a PID (proportion integration differentiation) controller in the PCR amplification instrument to be in a non-working state, and simultaneously transmitting the temperature difference and the temperature difference variation to the first fuzzy controller so as to control the magnitude of a driving current output by a power supply in the PCR amplification instrument to a lifting temperature regulator through the first fuzzy controller, wherein the temperature difference variation is equal to the difference between the temperature difference and the temperature difference corresponding to the previous sampling time node;
if not, controlling the first fuzzy controller to be in a non-working state, controlling the second fuzzy controller and the PID controller to be in a working state, simultaneously transmitting the temperature difference and the temperature difference variation to the second fuzzy controller, and transmitting the temperature difference to the PID controller, so as to adjust the control parameter of the PID controller through the second fuzzy controller, and finally controlling the magnitude of the driving current output by the power supply to the temperature rising and falling adjuster through the PID controller.
Based on the above invention, a temperature control scheme applied to a PCR amplification instrument and adopting a multi-mode segment control strategy is provided, i.e. a comparison result of a real-time target temperature difference and a preset temperature difference threshold is used as a decision basis for switching between a fuzzy control mode and a fuzzy PID control mode, the fuzzy control mode can be applied when the target temperature difference is large, so as to ensure that the temperature is increased/decreased at the fastest speed, and the fuzzy PID control mode is applied when the target temperature difference is small, so as to ensure low steady-state error and low overshoot, so as to achieve the purpose of accurately maintaining the temperature, further enable the temperature control system of the PCR amplification instrument to have the multi-mode temperature control mode, enable the system performance to achieve high temperature increase/decrease speed, low overshoot, small oscillation and high-precision unification to the greatest extent, improve the amplification efficiency of DNA fragments, shorten the time required for reaction, and facilitate to obtain the PCR detection result quickly, is convenient for practical application and popularization.
In one possible design, when the current sampling time node is in the middle of the temperature maintaining phase, the temperature control method further includes:
and when the absolute value of the temperature difference is judged to be less than or equal to the preset temperature difference threshold value, controlling the PID controller to be in a working state, controlling the first fuzzy controller and the second fuzzy controller to be in a non-working state, and simultaneously transmitting the temperature difference to the PID controller so as to control the magnitude of the driving current output by the power supply to the temperature rising and falling regulator through the PID controller.
In one possible design, acquiring a real-time temperature value acquired by a temperature sensor in the PCR amplifier at a current sampling time node includes:
acquiring temperature values continuously acquired by at least three temperature sensors at the current sampling time node for multiple times, wherein the at least three temperature sensors are respectively arranged at different sampling point positions;
and carrying out arithmetic mean filtering processing on all temperature values continuously and repeatedly acquired at the current sampling time node to obtain the real-time temperature value.
In one possible design, controlling a first fuzzy controller in the PCR amplifier to be in an operating state, and controlling a second fuzzy controller and a PID controller in the PCR amplifier to be in a non-operating state comprises:
and sending a switch-on instruction to a first switch electrically connected at the output end of the first fuzzy controller, and sending a switch-off instruction to a second switch electrically connected at the output end of the PID controller so as to set the output digital signal of the PID controller to zero.
In one possible design, controlling the first fuzzy controller to be in a non-operating state and controlling the second fuzzy controller and the PID controller to be in an operating state includes:
and sending an off instruction to a first switch electrically connected at the output end of the first fuzzy controller, and sending an on instruction to a second switch electrically connected at the output end of the PID controller so as to set the output digital signal of the first fuzzy controller to zero.
In one possible design, the control parameters include a proportional gain, an integral gain, and a differential gain of the PID controller; the membership function in the second fuzzy controller adopts a triangular membership function; all domains in the second fuzzy controller are defined as [ -X, X ], and the domain range of the temperature difference value is finely adjusted by a first quantization factor corresponding to a set value of 2, the domain range of the temperature difference value variation amount is finely adjusted by a second quantization factor corresponding to a set value of 2.5, the domain range of the proportional gain correction amount is finely adjusted by a third quantization factor corresponding to a set value of 10, the domain range of the integral gain correction amount is finely adjusted by a fourth quantization factor corresponding to a set value of 10, and the domain range of the differential gain correction amount is finely adjusted by a fifth quantization factor corresponding to a set value of 1, where X represents a positive integer equal to 6, and the set value represents a product of the corresponding quantization factor and the positive integer X.
In a second aspect, the invention provides a temperature control device of a PCR amplification instrument, which is arranged in a controller of the PCR amplification instrument and comprises a temperature acquisition unit, a temperature difference calculation unit, a comparison and judgment unit, a device control unit, a first transmission unit and a second transmission unit;
the temperature acquisition unit is used for acquiring a real-time temperature value acquired by a temperature sensor in the PCR amplification instrument at a current sampling time node;
the temperature difference calculating unit is in communication connection with the temperature acquiring unit and is used for calculating and obtaining a temperature difference value between the real-time temperature value and a target temperature value, wherein the target temperature value corresponds to the current sampling time node;
the comparison and judgment unit is in communication connection with the temperature difference calculation unit and is used for judging whether the absolute value of the temperature difference value is greater than a preset temperature difference threshold value, wherein the preset temperature difference threshold value is between 2 and 3 ℃;
the device control unit is in communication connection with the comparison and judgment unit and is used for controlling a first fuzzy controller in the PCR amplification instrument to be in a working state and controlling a second fuzzy controller and a PID controller in the PCR amplification instrument to be in a non-working state when the absolute value is judged to be larger than the preset temperature difference threshold value, and controlling the first fuzzy controller to be in the non-working state and controlling the second fuzzy controller and the PID controller to be in the working state when the absolute value is judged to be smaller than or equal to the preset temperature difference threshold value;
the first transmission unit is respectively in communication connection with the device control unit and the temperature difference calculation unit, and is used for transmitting the temperature difference value and the temperature difference value variation to the first fuzzy controller when the first fuzzy controller is controlled to be in a working state, so that the magnitude of the driving current output from the power supply in the PCR amplification instrument to the temperature rising and falling regulator is controlled through the first fuzzy controller, wherein the temperature difference value variation is equal to the difference between the temperature difference value and the temperature difference value corresponding to the previous sampling time node;
the second transmission unit is respectively in communication connection with the device control unit and the temperature difference calculation unit, and is used for transmitting the temperature difference value and the temperature difference value variation to the second fuzzy controller and transmitting the temperature difference value to the PID controller when the second fuzzy controller and the PID controller are controlled to be in working states, so that the control parameters of the PID controller are adjusted through the second fuzzy controller, and finally the driving current output by the power supply to the lifting temperature adjuster is controlled through the PID controller.
In a third aspect, the present invention provides a temperature control system for a PCR amplification apparatus, comprising a controller, a temperature sensor, a first fuzzy controller, a second fuzzy controller, a PID controller, a power supply, a current regulator, and a temperature raising/lowering regulator, wherein the controller is configured to perform the temperature control method according to the first aspect or any one of the possible designs of the first aspect;
the controller is respectively in communication connection with the temperature sensor, the first fuzzy controller, the second fuzzy controller and the PID controller, wherein the output end of the first fuzzy controller and the output end of the PID controller are respectively in communication connection with the controlled end of the current regulator, and the output end of the second fuzzy controller is in communication connection with the PID controller;
the power supply is electrically connected with the current input end of the current regulator, and the current output end of the current regulator is electrically connected with the lifting temperature regulator.
In a fourth aspect, the present invention provides a computer device, comprising a memory, a processor and a transceiver, which are sequentially connected in communication, wherein the memory is used for storing a computer program, the transceiver is used for transceiving a message, and the processor is used for reading the computer program and executing the temperature control method according to the first aspect or any one of the possible designs of the first aspect.
In a fifth aspect, the present invention provides a computer-readable storage medium having stored thereon instructions which, when executed on a computer, perform the temperature control method as described in the first aspect or any one of the possible designs of the first aspect.
In a sixth aspect, the present invention provides a computer program product containing instructions which, when run on a computer, cause the computer to perform the temperature control method as described above in the first aspect or any one of the possible designs of the first aspect.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly described below, it is obvious that the drawings in the following description are only some embodiments of the present invention, and for those skilled in the art, other drawings can be obtained according to the drawings without creative efforts.
FIG. 1 is a schematic structural diagram of a temperature control system of a PCR amplification apparatus provided by the present invention.
FIG. 2 is a schematic flow chart of the temperature control method of the PCR amplification instrument provided by the present invention.
FIG. 3 is a schematic diagram of the PID control algorithm provided by the present invention.
FIG. 4 is a diagram of an example of a simulation structure for implementing a PID control algorithm in MATLAB software according to the present invention.
FIG. 5 is a diagram of an example of a simulation structure for implementing a fuzzy PID control algorithm in MATLAB software according to the present invention.
FIG. 6 is a diagram of an example of a simulation structure for implementing the temperature control method in MATLAB software.
FIG. 7 is an exemplary graph of temperature change obtained by simulation in MATLAB software using the temperature control method provided by the present invention.
FIG. 8 is an exemplary graph of temperature change at a temperature of about 58 ℃ after the temperature control method is adopted in MATLAB software.
FIG. 9 is an exemplary graph of temperature change at a temperature of about 98 ℃ after the temperature control method is adopted in MATLAB software.
FIG. 10 is an exemplary graph of temperature change at a temperature of about 58 ℃ after simulation in MATLAB software using the temperature control method provided by the present invention.
FIG. 11 is a schematic structural diagram of a temperature control device of the PCR amplification apparatus provided by the present invention.
Fig. 12 is a schematic structural diagram of a computer device provided by the present invention.
Detailed Description
The invention is further described with reference to the following figures and specific embodiments. It should be noted that the description of the embodiments is provided to help understanding of the present invention, but the present invention is not limited thereto. Specific structural and functional details disclosed herein are merely representative of exemplary embodiments of the invention. This invention may, however, be embodied in many alternate forms and should not be construed as limited to the embodiments set forth herein.
It will be understood that, although the terms first, second, etc. may be used herein to describe various objects, these objects should not be limited by these terms. These terms are only used to distinguish one object from another. For example, a first object may be referred to as a second object, and similarly, a second object may be referred to as a first object, without departing from the scope of example embodiments of the present invention.
It should be understood that, for the term "and/or" as may appear herein, it is merely an associative relationship that describes an associated object, meaning that three relationships may exist, e.g., a and/or B may mean: a exists alone, B exists alone or A and B exist at the same time; for the term "/and" as may appear herein, which describes another associative object relationship, it means that two relationships may exist, e.g., a/and B, may mean: a exists singly or A and B exist simultaneously; in addition, for the character "/" that may appear herein, it generally means that the former and latter associated objects are in an "or" relationship.
As shown in fig. 1 to 3, the temperature control method for a PCR amplification instrument provided in the first aspect of this embodiment is executed by a controller in the PCR amplification instrument, so that a temperature control system of the PCR amplification instrument has a multi-modal temperature control mode, that is, the temperature control system can be flexibly switched between a fuzzy control mode and a fuzzy PID control mode, thereby ensuring that temperature rise/fall can be performed at the fastest speed when a target temperature difference is large, and also ensuring that a temperature can be accurately maintained when the target temperature difference is small, thereby improving amplification efficiency of DNA fragments, shortening time required for a reaction, and facilitating to quickly obtain a PCR detection result.
As shown in fig. 1, the temperature control system of the PCR amplification apparatus includes, but is not limited to, a controller, a temperature sensor, a first fuzzy controller, a second fuzzy controller, a PID controller, a power supply, a current regulator, and a temperature regulator, wherein the controller is used for executing a specific temperature control method; the controller is respectively in communication connection with the temperature sensor, the first fuzzy controller, the second fuzzy controller and the PID controller, wherein the output end of the first fuzzy controller and the output end of the PID controller are respectively in communication connection with the controlled end of the current regulator, and the output end of the second fuzzy controller is in communication connection with the PID controller; the power supply is electrically connected with the current input end of the current regulator, and the current output end of the current regulator is electrically connected with the lifting temperature regulator.
In a specific structure of the temperature control system, the controller is a core component for temperature control, which can be implemented by, but not limited to, a microprocessor chip based on the STM32F103 series and its peripheral circuits. The temperature sensor is used for collecting sample temperature values, the number of the temperature sensors can be multiple, so that the purpose of simultaneously measuring the temperature at multiple different sampling points can be realized, and the temperature sensor can be realized by adopting a conventional temperature sensor. The first fuzzy controller and the second fuzzy controller are respectively conventional algorithm modules for realizing a fuzzy control algorithm (i.e. a control method utilizing basic ideas and theories of fuzzy mathematics), and can be respectively a hardware module or a software module with an independent corresponding function, wherein the first fuzzy controller is used for outputting a digital signal capable of generating a control action to the current regulator through the fuzzy control algorithm so as to control the magnitude of the driving current output by the power supply to the lifting temperature regulator and realize the regulation purpose of heating or cooling; the second fuzzy controller is used for adjusting and correcting the control parameters of the PID controller through a fuzzy control algorithm so as to control the magnitude of the driving current output by the power supply to the lifting temperature regulator through the PID controller. The PID controller is a conventional algorithm module for realizing a PID control algorithm (namely, a control algorithm which combines three links of proportion, integral and differentiation into a whole, is the most mature control algorithm with the most extensive application in the technology in a continuous system, appears in 30 to 40 years of the 20 th century and is suitable for occasions with unclear knowledge on a controlled object model), and can also be a hardware module or a software module with an independent corresponding function, and the PID controller is used for outputting a digital signal capable of generating a control action to the current regulator through the PID control algorithm so as to control the magnitude of the driving current output by the power supply to the lifting temperature regulator and realize the regulation purpose of heating or cooling. The power supply is used for providing electric energy support for the lifting temperature regulator through the current regulator, and is preferably a direct current power supply. The current regulator is used for realizing the purpose of dynamically regulating the magnitude of the passed current under the control of the first fuzzy controller or the PID controller, and can be realized by adopting a numerical control programmable resistor without limitation. The temperature rising and falling adjuster is used for achieving the purpose of heating or radiating and cooling a sample, and can be but not limited to an electric heater (namely used for heating), a fan (namely used for radiating and cooling), a semiconductor refrigeration piece (namely used for heating and cooling by changing the current direction, and can be used for heating and cooling as well as radiating and cooling) and the like.
As shown in FIG. 2, the temperature control method of the PCR amplification apparatus may include, but is not limited to, the following steps S1-S5.
S1, acquiring a real-time temperature value acquired by a temperature sensor in the PCR amplification instrument at a current sampling time node.
In step S1, in order to improve the accuracy of temperature sampling, it is necessary to perform digital filtering processing on the signal after temperature sampling. Therefore, it is preferable, but not limited to, that the following steps S11 to S12 are included: s11, temperature values continuously and repeatedly acquired by at least three temperature sensors at the current sampling time node are acquired, wherein the at least three temperature sensors are respectively arranged at different sampling point positions; and S12, carrying out arithmetic mean filtering processing on all temperature values continuously and repeatedly acquired at the current sampling time node to obtain the real-time temperature value. In the foregoing step S11, if the temperature sampling frequency is 100Hz according to the requirement of the temperature control system, 0.1 second may be used as the time interval between two adjacent sampling time nodes, M temperature sensors are applied when the sampling time nodes arrive, N temperature values are obtained by continuously collecting N times based on the temperature sampling frequency (i.e., collecting once every 0.01 second), and then an arithmetic mean filtering process is performed on M × N temperature values to obtain the final real-time temperature value from which the random interference noise in the temperature sampling is eliminated, where M and N respectively represent positive integers greater than 2 and less than 10, that is, M sampling points. In addition, the specific manner of the arithmetic mean filtering processing is the conventional digital filtering processing manner, which means that a signal is continuously sampled for a plurality of times at a certain time (i.e. a sampling time node), and then the sampled values are arithmetically averaged to be used as the signal value at the time, wherein the number of times of continuous sampling is determined as the specific case may be.
And S2, calculating to obtain a temperature difference value between the real-time temperature value and a target temperature value, wherein the target temperature value corresponds to the current sampling time node.
In step S2, the target temperature value needs to be dynamically determined according to the time period, and in a typical PCR amplification reaction cycle, the target temperature value is mainly divided into three time periods of 58 ℃, 78 ℃ and 98 ℃, i.e. the temperature is raised from room temperature to 58 ℃ and is kept for 30 seconds; raising the temperature from 58 ℃ to 98 ℃, and keeping the temperature for 30 seconds; reducing the temperature from 98 ℃ to 58 ℃ and keeping the temperature for 30 seconds; raising the temperature from 58 ℃ to 78 ℃ and keeping the temperature for 30 seconds; raising the temperature from 78 ℃ to 98 ℃, and keeping for 30 seconds; the temperature is then cycled sequentially from 98 deg.C to 58 deg.C to 78 deg.C. Thus, in the PCR amplification apparatus, the target temperature value may be 58 ℃, 78 ℃ or 98 ℃ in the temperature increasing process period, or 58 ℃ in the temperature decreasing process period, or 58 ℃, 78 ℃ or 98 ℃ in the temperature maintaining period, or the like.
S3, judging whether the absolute value of the temperature difference value is larger than a preset temperature difference threshold value, wherein the preset temperature difference threshold value is between 2 and 3 ℃.
In the step S3, the preset temperature difference threshold is used as a decision basis for switching between the fuzzy control mode and the fuzzy PID control mode, and the specific value thereof is preferably 2.5 ℃.
And S4, if so, controlling a first fuzzy controller in the PCR amplification instrument to be in a working state, controlling a second fuzzy controller and a PID (proportion integration differentiation) controller in the PCR amplification instrument to be in a non-working state, and simultaneously transmitting the temperature difference and the temperature difference variation to the first fuzzy controller so as to control the magnitude of a driving current output by a power supply in the PCR amplification instrument to a lifting temperature regulator through the first fuzzy controller, wherein the temperature difference variation is equal to the difference between the temperature difference and the temperature difference corresponding to the previous sampling time node.
In step S4, if it is determined that the absolute value of the temperature difference is greater than the preset temperature difference threshold, it indicates that the actual temperature value is a certain distance away from the target temperature value at the initial stage of the temperature increasing/decreasing process, and considering that the temperature control system does not have any special requirement for the static performance of the temperature at this time, but needs the temperature increasing/decreasing speed as fast as possible, so that the output digital signal can be made as large as possible by the first fuzzy controller at this time, and thus, the power supply outputs a large driving current to the temperature increasing/decreasing regulator through the adjustment and control of the current regulator, so as to increase the heating/heat dissipation speed, and achieve the purpose of fast temperature increasing/decreasing by applying the fuzzy control mode. Further, the temperature difference value and the temperature difference value change amount, etc. are input parameters necessary in the fuzzy control algorithm for temperature control, and after the step S4 is performed, it will return to the step S1 to be performed when the next sampling time node arrives, so as to continuously perform temperature control.
In step S4, in order to avoid the uncertain digital signals output by the PID controller causing control interference to the current regulator, it is preferable to control the first fuzzy controller in the PCR amplifier to be in an operating state, and control the second fuzzy controller and the PID controller in the PCR amplifier to be in a non-operating state, including but not limited to: and sending a switch-on instruction to a first switch electrically connected at the output end of the first fuzzy controller, and sending a switch-off instruction to a second switch electrically connected at the output end of the PID controller so as to set the output digital signal of the PID controller to zero. The first switch and the second switch may be physical electrically-controlled switches (e.g., relay switches), or virtual switches implementing a switching function, which are connected in series between the corresponding controller and the current regulator, and communicatively connect the controlled end to the controller, so as to receive a command, and enable a digital signal output by the corresponding controller to be transmitted to the current regulator when the switch is turned on, thereby performing an action of dynamically adjusting the magnitude of the driving current, and disable the digital signal output by the corresponding controller when the switch is turned off, thereby performing an action of setting the magnitude of the driving current to zero, and not performing the action of adjusting the magnitude of the driving current. Therefore, the aim that the whole temperature control system only works in the fuzzy control mode can be fulfilled by sending an on command to the first switch and sending an off command to the second switch.
And S5, if not, controlling the first fuzzy controller to be in a non-working state, controlling the second fuzzy controller and the PID controller to be in a working state, simultaneously transmitting the temperature difference and the temperature difference variation to the second fuzzy controller, and transmitting the temperature difference to the PID controller, so that the control parameters of the PID controller are adjusted through the second fuzzy controller, and finally controlling the driving current output by the power supply to the lifting temperature adjuster through the PID controller.
In the step S5, if it is determined that the absolute value of the temperature difference is less than or equal to the preset temperature difference threshold, it indicates that the actual temperature value is close to the target temperature value at the end of the temperature rising/lowering process or during the temperature maintaining process, and in order to meet the static performance requirement of the temperature control system, the control parameters of the PID controller may be properly adjusted by using the second fuzzy controller according to the change of the input parameters (i.e., the temperature difference variation, etc.), which not only can make up the disadvantage of the PID controller in poor dynamic performance, but also can effectively control the overshoot of the system, thereby achieving the purpose of accurately maintaining the temperature by applying the fuzzy PID control mode. Further, the temperature difference value variation amount, and the like are input parameters necessary in the fuzzy control algorithm for temperature control, the temperature difference value is input parameter necessary in the PID control algorithm for temperature control, and after the step S5 is performed, it will return to the step S1 to be performed when the next sampling time node arrives, so as to continuously perform temperature control.
In step S5, the operating principle of the PID controller is as shown in fig. 3, and the transfer function gc (S) thereof can be expressed as follows:
in the formula, s represents a variable, kpDenotes the proportional gain, kiRepresenting the integral gain, kdThe differential gain is expressed, i.e. by adjusting the control parameter: proportional gain kpIntegral gain kiAnd a differential gain kdThe controlled object has better dynamic response and static response, thereby meeting the requirements of the control system. The specific working process of the PID controller comprises a proportion link, an integral link and a differential link, wherein the proportion link is used for adjusting system control quantity in proportion according to deviation quantity so as to generate a control effect and reduce the deviation, namely the proportion gain has the effect of increasing the system response speed, the system response speed is higher when the proportion gain is larger, but the system is easy to generate overshoot, the system adjustment precision is influenced when the proportion gain is too small, and the system response time is prolonged, so that the dynamic response of the system is poor; the integral link is used for eliminating static difference and improving the zero-difference degree of the system, the integral time constant (namely integral gain) of the integral link determines the action strength of the integral link, but the stability of the system is influenced if the integral action is too strong; the differential link is used for adjusting the system control quantity according to the variation trend of the deviation quantity and is introduced in advance before the deviation signal is greatly changedA correction signal is used to increase the system action speed and reduce the adjustment time, and the adjustment of the differential parameter (i.e. the differential gain) needs to be paid attention to that the system oscillation is caused by too strong differential action. In this embodiment, for convenience of debugging in the temperature control system, the control parameters of the PID controller are: proportional gain kpIntegral gain kiAnd a differential gain kdRespectively defined as external adjustable input parameters, namely a simulation structure for realizing a PID control algorithm as shown in FIG. 4, wherein ET represents a temperature difference value corresponding to the current sampling time node; the icon 3-kp1 represents the proportional gain kp(ii) a The icon 4-ki1 represents the integral gain ki(ii) a The icon 5-kd1 represents the differential gain kd(ii) a Icons fcn _2, fcn _3 and fcn _4 represent the magnitudes of the output contribution values of the proportional element, the integral element and the differential element after the PID control algorithm is solved.
In step S5, the output u (t) of the second fuzzy controller can be expressed by the following formula:
wherein t represents a time variable, KPIndicating an initial proportional gain, K, set by the PID controllerIIndicating the initial integral gain, K, set by the PID controllerDRepresenting the initial derivative gain set by the PID controller. Δ KP、ΔKIAnd Δ KDOutput quantities which are respectively output by the second fuzzy controller and correspond to three control parameters of the PID controller one by one are used for respectively correcting proportional gain, integral gain and differential gain of the PID controller, e (t) represents a temperature difference value corresponding to a time variable t, de (t) represents a temperature difference value variation corresponding to the time variable t, and dt represents a differential form of the time variable t. The second fuzzy controller is mainly used for finding out a fuzzy relation corresponding to correction quantities of three control parameters of the PID controller, a temperature difference value e (t) and a temperature difference value change de (t), and further self-tuning a parameter delta KP、ΔKIAnd Δ KDAnd continuously detecting the temperature difference e (t) and the temperature difference change de (t) during operation, and performing online correction on the three control parameters of the PID controller to meet different requirements on parameters of the PID controller at different stages of temperature change, so that a controlled object (namely the lifting temperature regulator) has good dynamic and static performances.
Specifically, the fuzzification input by the temperature control system is fuzzification of a temperature difference value and a temperature difference value variation, and in the second fuzzy controller, the membership functions are all triangular membership functions, that is, the membership of the fuzzification of the system variable is calculated according to the triangular membership functions. Since in the PID controller, the proportional gain kpRelated to the response speed of the system, kpThe larger the response speed of the system, the higher the system accuracy, but the larger the kpThe system is easy to generate overshoot and even unstable; the integral gain kiFor eliminating steady-state error of system, kiThe larger the steady state temperature difference of the system, the faster it is eliminated, but the too large kiIntegral saturation can occur, causing large overshoot and excessive oscillation, so that the proportional gain k needs to be reasonably balanced through continuous operation and trial in the temperature control systempIntegral gain kiAnd a differential gain kdThe relationship (2) can obtain a better control effect.
The simulation structure for implementing the fuzzy PID control algorithm, as shown in FIG. 5, has the proportional gain kpIntegral gain kiAnd a differential gain kdThe fuzzy control tables are respectively placed in icons tip-p2, tip-i2 and tip-d2 to facilitate quick calling of a program, e1 represents the temperature difference, edt1 represents the temperature difference variation, and the two quantities are input quantities; for ease of tuning, all domains of discourse in the second fuzzy controller may be defined as [ -X, X]And the universe of discourse of the temperature difference is finely adjusted by a first quantization factor corresponding to the set value of 2, the universe of discourse of the variation of the temperature difference is finely adjusted by a second quantization factor corresponding to the set value of 2.5, and the scale-up is finely adjusted by a third quantization factor corresponding to the set value of 10A domain range of the gain correction amount, wherein X represents a positive integer equal to 6, a domain range of the integral gain correction amount is fine-tuned by a fourth quantization factor corresponding to a set value of 10, and a domain range of the differential gain correction amount is fine-tuned by a fifth quantization factor corresponding to a set value of 1, wherein the set value represents a product of the corresponding quantization factor and the positive integer X; the icons 6-kp2, 7-ki2, and 8-kd2 refer to the initial values of proportional gain, integral gain, and derivative gain, respectively, in the PID controller; and the system functions MKP, MKI and MKD respectively represent the contribution values of a proportional link, an integral link and a differential link to output after the fuzzy control algorithm is solved.
In step S5, in order to avoid the uncertain digital signal output by the first fuzzy controller causing control interference to the current regulator, it is preferable to control the first fuzzy controller to be in a non-operating state, and control the second fuzzy controller and the PID controller to be in an operating state, including but not limited to: and sending an off instruction to a first switch electrically connected at the output end of the first fuzzy controller, and sending an on instruction to a second switch electrically connected at the output end of the PID controller so as to set the output digital signal of the first fuzzy controller to zero. For specific description of the first switch and the second switch, refer to the foregoing step S4, which is not described herein again.
As shown in fig. 6 to 10, based on the simulation structure for implementing the temperature control method as shown in fig. 6, the temperature change curves obtained by simulation as shown in fig. 7 to 10 can be obtained, and it can be seen that, in the temperature raising process from room temperature to 58 ℃, from 58 ℃ to 98 ℃, from 58 ℃ to 78 ℃ and from 78 ℃ to 98 ℃ and in the temperature lowering process from 98 ℃ to 58 ℃, rapid temperature raising or temperature lowering can be achieved, and after the temperature raising to 58 ℃ is stabilized, the steady-state error can be made to be less than 0.1 ℃, the overshoot can be made to be within 2 ℃, and after the temperature raising to 98 ℃ is stabilized, the steady-state error can be made to be less than 0.1 ℃, and after the temperature lowering to 58 ℃ is stabilized, the steady-state error can be made to be less than 0.1 ℃, and the error occasionally is between 0.1 ℃ and 0.15 ℃ after many cycles. In summary, the experimental data results show that the temperature control algorithm provided by the embodiment can realize the effects that the temperature rise steady-state error is less than 0.1 ℃ and the temperature drop error is less than 0.15 ℃ in precision, so that the amplification efficiency of the DNA fragments can be improved, the time required by the reaction can be shortened, and the PCR detection result can be obtained quickly.
Therefore, based on the temperature control method described in the foregoing steps S1-S5, a temperature control scheme applied to a PCR amplification apparatus and adopting a multi-mode segment control strategy is provided, that is, by using the comparison result of the real-time target temperature difference and the preset temperature difference threshold as the decision basis for switching between the fuzzy control mode and the fuzzy PID control mode, the fuzzy control mode can be applied when the target temperature difference is large, so as to ensure the fastest temperature rise/fall, and the fuzzy PID control mode is applied when the target temperature difference is small, so as to ensure the low steady-state error and the low overshoot, so as to achieve the purpose of accurately maintaining the temperature, and further, the temperature control system of the PCR amplification apparatus has the multi-mode temperature control mode, so that the system performance can achieve the unification of fast temperature rise and fall, low overshoot, small oscillation and high precision to the greatest extent, thereby improving the amplification efficiency of DNA fragments, shortening the time required for reaction, is beneficial to quickly obtaining the PCR detection result and is convenient for practical application and popularization.
On the basis of the technical solution of the first aspect, this embodiment further specifically provides a possible design that further reduces steady-state error and overshoot in the temperature maintenance process, that is, when the current sampling time node is in the middle stage of the temperature maintenance phase, the temperature control method further includes, but is not limited to: and when the absolute value of the temperature difference is judged to be less than or equal to the preset temperature difference threshold value, controlling the PID controller to be in a working state, controlling the first fuzzy controller and the second fuzzy controller to be in a non-working state, and simultaneously transmitting the temperature difference to the PID controller so as to control the magnitude of the driving current output by the power supply to the temperature rising and falling regulator through the PID controller. Because the range of temperature fluctuation caused by the residual heat quantity brought in the temperature rising process or the residual cold quantity brought in the temperature lowering process is very small when the current sampling time node is in the middle stage of the temperature maintaining stage, the temperature control is carried out by directly applying the PID control algorithm, the effects of smaller steady-state error and overshoot can be realized, and the aim of accurately maintaining the temperature is further fulfilled. The middle period may be, for example, 10 th to 20 th seconds out of 30 seconds.
Therefore, based on the possible design one described in detail above, the effect of smaller steady-state error and overshoot can be achieved in the middle stage of the temperature maintaining stage, and the purpose of accurately maintaining the temperature can be further achieved.
As shown in fig. 11, a second aspect of the present embodiment provides a virtual device for implementing the temperature control method according to any one of the first aspect or the first aspect, wherein the virtual device is disposed in a controller of the PCR amplification apparatus, and includes a temperature obtaining unit, a temperature difference calculating unit, a comparison and determination unit, a device control unit, a first transmission unit, and a second transmission unit;
the temperature acquisition unit is used for acquiring a real-time temperature value acquired by a temperature sensor in the PCR amplification instrument at a current sampling time node;
the temperature difference calculating unit is in communication connection with the temperature acquiring unit and is used for calculating and obtaining a temperature difference value between the real-time temperature value and a target temperature value, wherein the target temperature value corresponds to the current sampling time node;
the comparison and judgment unit is in communication connection with the temperature difference calculation unit and is used for judging whether the absolute value of the temperature difference value is greater than a preset temperature difference threshold value, wherein the preset temperature difference threshold value is between 2 and 3 ℃;
the device control unit is in communication connection with the comparison and judgment unit and is used for controlling a first fuzzy controller in the PCR amplification instrument to be in a working state and controlling a second fuzzy controller and a PID controller in the PCR amplification instrument to be in a non-working state when the absolute value is judged to be larger than the preset temperature difference threshold value, and controlling the first fuzzy controller to be in the non-working state and controlling the second fuzzy controller and the PID controller to be in the working state when the absolute value is judged to be smaller than or equal to the preset temperature difference threshold value;
the first transmission unit is respectively in communication connection with the device control unit and the temperature difference calculation unit, and is used for transmitting the temperature difference value and the temperature difference value variation to the first fuzzy controller when the first fuzzy controller is controlled to be in a working state, so that the magnitude of the driving current output from the power supply in the PCR amplification instrument to the temperature rising and falling regulator is controlled through the first fuzzy controller, wherein the temperature difference value variation is equal to the difference between the temperature difference value and the temperature difference value corresponding to the previous sampling time node;
the second transmission unit is respectively in communication connection with the device control unit and the temperature difference calculation unit, and is used for transmitting the temperature difference value and the temperature difference value variation to the second fuzzy controller and transmitting the temperature difference value to the PID controller when the second fuzzy controller and the PID controller are controlled to be in working states, so that the control parameters of the PID controller are adjusted through the second fuzzy controller, and finally the driving current output by the power supply to the lifting temperature adjuster is controlled through the PID controller.
For the working process, working details and technical effects of the foregoing device provided in the second aspect of this embodiment, reference may be made to the temperature control method in any one of the first aspect and the first aspect, which is not described herein again.
As shown in fig. 1, a third aspect of the present embodiment provides a temperature control system adopting any one of the possible designs of the first aspect or the first aspect, including a controller, a temperature sensor, a first fuzzy controller, a second fuzzy controller, a PID controller, a power supply, a current regulator, and a lift temperature regulator, wherein the controller is configured to execute the temperature control method according to any one of the possible designs of the first aspect or the first aspect;
the controller is respectively in communication connection with the temperature sensor, the first fuzzy controller, the second fuzzy controller and the PID controller, wherein the output end of the first fuzzy controller and the output end of the PID controller are respectively in communication connection with the controlled end of the current regulator, and the output end of the second fuzzy controller is in communication connection with the PID controller;
the power supply is electrically connected with the current input end of the current regulator, and the current output end of the current regulator is electrically connected with the lifting temperature regulator.
For the working process, working details and technical effects of the foregoing system provided in the third aspect of this embodiment, reference may be made to the first aspect or any one of the temperature control methods that may be designed in the first aspect, which is not described herein again.
As shown in fig. 12, a fourth aspect of this embodiment provides a computer device for executing the temperature control method according to any one of the first aspect or the possible designs of the first aspect, and includes a memory, a processor, and a transceiver, which are sequentially connected in a communication manner, where the memory is used for storing a computer program, the transceiver is used for transceiving a message, and the processor is used for reading the computer program to execute the temperature control method according to any one of the first aspect or the possible designs of the first aspect. For example, the Memory may include, but is not limited to, a Random-Access Memory (RAM), a Read-Only Memory (ROM), a Flash Memory (Flash Memory), a First-in First-out (FIFO), and/or a First-in Last-out (FILO), and the like; the transceiver may be, but is not limited to, a WiFi (wireless fidelity) wireless transceiver, a bluetooth wireless transceiver, a GPRS (General Packet Radio Service) wireless transceiver, and/or a ZigBee (ZigBee protocol, low power consumption local area network protocol based on ieee802.15.4 standard) wireless transceiver, etc.; the processor may be, but is not limited to, a microprocessor of the model number STM32F105 family. In addition, the computer device may also include, but is not limited to, a power module, a display screen, and other necessary components.
For the working process, working details, and technical effects of the foregoing computer device provided in the fourth aspect of this embodiment, reference may be made to the first aspect or any one of the possible designs of the temperature control method in the first aspect, which is not described herein again.
A fifth aspect of the present embodiment provides a computer-readable storage medium storing instructions including any one of the first aspect or any one of the possible designs of the temperature control method of the first aspect, that is, the computer-readable storage medium has instructions stored thereon, and when the instructions are executed on a computer, the temperature control method of any one of the possible designs of the first aspect or the first aspect is executed. The computer-readable storage medium refers to a carrier for storing data, and may include, but is not limited to, floppy disks, optical disks, hard disks, flash memories, flash disks and/or Memory sticks (Memory sticks), etc., and the computer may be a general-purpose computer, a special-purpose computer, a computer network, or other programmable devices.
For the working process, the working details and the technical effects of the foregoing computer-readable storage medium provided in the fifth aspect of this embodiment, reference may be made to the first aspect or any one of the possible designs of the temperature control method in the first aspect, which is not described herein again.
A sixth aspect of the present embodiment provides a computer program product containing instructions which, when executed on a computer, cause the computer to perform the temperature control method according to the first aspect or any one of the possible designs of the first aspect. The computer may be a general purpose computer, a special purpose computer, a network of computers, or other programmable devices.
Finally, it should be noted that the present invention is not limited to the above alternative embodiments, and that various other forms of products can be obtained by anyone in light of the present invention. The above detailed description should not be taken as limiting the scope of the invention, which is defined in the claims, and which the description is intended to be interpreted accordingly.
Claims (10)
1. A temperature control method of a PCR amplification instrument, which is executed by a controller in the PCR amplification instrument, is characterized by comprising the following steps:
acquiring a real-time temperature value acquired by a temperature sensor in the PCR amplification instrument at a current sampling time node;
calculating to obtain a temperature difference value between the real-time temperature value and a target temperature value, wherein the target temperature value corresponds to the current sampling time node;
judging whether the absolute value of the temperature difference value is larger than a preset temperature difference threshold value, wherein the preset temperature difference threshold value is between 2 and 3 ℃;
if yes, controlling a first fuzzy controller in the PCR amplification instrument to be in a working state, controlling a second fuzzy controller and a PID (proportion integration differentiation) controller in the PCR amplification instrument to be in a non-working state, and simultaneously transmitting the temperature difference and the temperature difference variation to the first fuzzy controller so as to control the magnitude of a driving current output by a power supply in the PCR amplification instrument to a lifting temperature regulator through the first fuzzy controller, wherein the temperature difference variation is equal to the difference between the temperature difference and the temperature difference corresponding to the previous sampling time node;
if not, controlling the first fuzzy controller to be in a non-working state, controlling the second fuzzy controller and the PID controller to be in a working state, simultaneously transmitting the temperature difference and the temperature difference variation to the second fuzzy controller, and transmitting the temperature difference to the PID controller, so as to adjust the control parameter of the PID controller through the second fuzzy controller, and finally controlling the magnitude of the driving current output by the power supply to the temperature rising and falling adjuster through the PID controller.
2. The temperature control method according to claim 1, wherein when the current sampling time node is in a middle stage of a temperature maintenance phase, the temperature control method further comprises:
and when the absolute value of the temperature difference is judged to be less than or equal to the preset temperature difference threshold value, controlling the PID controller to be in a working state, controlling the first fuzzy controller and the second fuzzy controller to be in a non-working state, and simultaneously transmitting the temperature difference to the PID controller so as to control the magnitude of the driving current output by the power supply to the temperature rising and falling regulator through the PID controller.
3. The temperature control method of claim 1, wherein obtaining the real-time temperature value collected by the temperature sensor in the PCR amplifier at the current sampling time node comprises:
acquiring temperature values continuously acquired by at least three temperature sensors at the current sampling time node for multiple times, wherein the at least three temperature sensors are respectively arranged at different sampling point positions;
and carrying out arithmetic mean filtering processing on all temperature values continuously and repeatedly acquired at the current sampling time node to obtain the real-time temperature value.
4. The temperature control method of claim 1, wherein controlling a first fuzzy controller in the PCR amplifier to be in an active state and controlling a second fuzzy controller and a PID controller in the PCR amplifier to be in an inactive state comprises:
and sending a switch-on instruction to a first switch electrically connected at the output end of the first fuzzy controller, and sending a switch-off instruction to a second switch electrically connected at the output end of the PID controller so as to set the output digital signal of the PID controller to zero.
5. The method of temperature control according to claim 1, wherein controlling the first fuzzy controller to be in a non-operational state and controlling the second fuzzy controller and the PID controller to be in an operational state comprises:
and sending an off instruction to a first switch electrically connected at the output end of the first fuzzy controller, and sending an on instruction to a second switch electrically connected at the output end of the PID controller so as to set the output digital signal of the first fuzzy controller to zero.
6. The temperature control method according to claim 1, wherein the control parameters include a proportional gain, an integral gain, and a differential gain of the PID controller; the membership function in the second fuzzy controller adopts a triangular membership function; all domains in the second fuzzy controller are defined as [ -X, X ], and the domain range of the temperature difference value is finely adjusted by a first quantization factor corresponding to a set value of 2, the domain range of the temperature difference value variation amount is finely adjusted by a second quantization factor corresponding to a set value of 2.5, the domain range of the proportional gain correction amount is finely adjusted by a third quantization factor corresponding to a set value of 10, the domain range of the integral gain correction amount is finely adjusted by a fourth quantization factor corresponding to a set value of 10, and the domain range of the differential gain correction amount is finely adjusted by a fifth quantization factor corresponding to a set value of 1, where X represents a positive integer equal to 6, and the set value represents a product of the corresponding quantization factor and the positive integer X.
7. A temperature control device of a PCR amplification instrument is arranged in a controller of the PCR amplification instrument and is characterized by comprising a temperature acquisition unit, a temperature difference calculation unit, a comparison and judgment unit, a device control unit, a first transmission unit and a second transmission unit;
the temperature acquisition unit is used for acquiring a real-time temperature value acquired by a temperature sensor in the PCR amplification instrument at a current sampling time node;
the temperature difference calculating unit is in communication connection with the temperature acquiring unit and is used for calculating and obtaining a temperature difference value between the real-time temperature value and a target temperature value, wherein the target temperature value corresponds to the current sampling time node;
the comparison and judgment unit is in communication connection with the temperature difference calculation unit and is used for judging whether the absolute value of the temperature difference value is greater than a preset temperature difference threshold value, wherein the preset temperature difference threshold value is between 2 and 3 ℃;
the device control unit is in communication connection with the comparison and judgment unit and is used for controlling a first fuzzy controller in the PCR amplification instrument to be in a working state and controlling a second fuzzy controller and a PID controller in the PCR amplification instrument to be in a non-working state when the absolute value is judged to be larger than the preset temperature difference threshold value, and controlling the first fuzzy controller to be in the non-working state and controlling the second fuzzy controller and the PID controller to be in the working state when the absolute value is judged to be smaller than or equal to the preset temperature difference threshold value;
the first transmission unit is respectively in communication connection with the device control unit and the temperature difference calculation unit, and is used for transmitting the temperature difference value and the temperature difference value variation to the first fuzzy controller when the first fuzzy controller is controlled to be in a working state, so that the magnitude of the driving current output from the power supply in the PCR amplification instrument to the temperature rising and falling regulator is controlled through the first fuzzy controller, wherein the temperature difference value variation is equal to the difference between the temperature difference value and the temperature difference value corresponding to the previous sampling time node;
the second transmission unit is respectively in communication connection with the device control unit and the temperature difference calculation unit, and is used for transmitting the temperature difference value and the temperature difference value variation to the second fuzzy controller and transmitting the temperature difference value to the PID controller when the second fuzzy controller and the PID controller are controlled to be in working states, so that the control parameters of the PID controller are adjusted through the second fuzzy controller, and finally the driving current output by the power supply to the lifting temperature adjuster is controlled through the PID controller.
8. A temperature control system of a PCR amplification instrument is characterized by comprising a controller, a temperature sensor, a first fuzzy controller, a second fuzzy controller, a PID controller, a power supply, a current regulator and a lifting temperature regulator, wherein the controller is used for executing the temperature control method according to any one of claims 1-6;
the controller is respectively in communication connection with the temperature sensor, the first fuzzy controller, the second fuzzy controller and the PID controller, wherein the output end of the first fuzzy controller and the output end of the PID controller are respectively in communication connection with the controlled end of the current regulator, and the output end of the second fuzzy controller is in communication connection with the PID controller;
the power supply is electrically connected with the current input end of the current regulator, and the current output end of the current regulator is electrically connected with the lifting temperature regulator.
9. A computer device, comprising a memory, a processor and a transceiver, which are connected in sequence in communication, wherein the memory is used for storing a computer program, the transceiver is used for transmitting and receiving messages, and the processor is used for reading the computer program and executing the temperature control method according to any one of claims 1-6.
10. A computer-readable storage medium having stored thereon instructions for performing the temperature control method according to any one of claims 1 to 6 when the instructions are run on a computer.
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Application publication date: 20211008 |