CN109957494B - Temperature control method for nucleic acid amplification instrument - Google Patents
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Abstract
The invention provides a temperature control method for a nucleic acid amplification instrument, which comprises an input device, a temperature sensor group, a chip, a primary refrigerating sheet, a secondary refrigerating sheet, a heat dissipation module and a PID control unit, wherein the input device is used for inputting a temperature signal; the temperature sensor group comprises a first temperature sensor, a second temperature sensor, a third temperature sensor and a fourth temperature sensor; the PID control unit comprises a temperature signal processing unit, a fuzzy operator and a PID controller, and the fuzzy operator is electrically connected with the PID controller; the temperature signal processing unit is electrically connected with the PID controller; the PID control unit makes a decision according to the results of the fuzzy operator and the PID controller, and controls the first-stage refrigerating sheet, the second-stage refrigerating sheet and the heat dissipation module to work; the temperature control method for the nucleic acid amplification instrument provided by the invention enables the temperature regulation reaction to be rapid and high in accuracy.
Description
Technical Field
The invention relates to the technical field of nucleic acid amplification instruments, in particular to a temperature control method for a nucleic acid amplification instrument.
Background
The nucleic acid amplification technology, especially the digital Polymerase Chain Reaction (PCR) technology, is a molecular biological technology for amplifying specific DNA molecular fragment, which uses DNA molecule as template and uses a pair of artificially synthesized specific oligonucleotide primers to quickly amplify the specific DNA molecular fragment by means of DNA Polymerase enzymatic Reaction, and has extremely important action in biology. The basic process of PCR reaction is divided into three steps. Firstly, DNA denaturation (94 ℃) is carried out, and a double-stranded DNA template is broken by hydrogen bonds under the action of heat to form single-stranded DNA; step two, annealing (55 ℃), reducing the temperature of the system, and combining the primer and the DNA template to form a local double strand; and step three, extending (72 ℃) to synthesize a DNA chain complementary with the template by taking dNTP as a raw material to extend from the 5 end to the 3 end of the primer under the action of Taq enzyme. The PCR instrument is used for performing denaturation, annealing and polymerization treatment on the amplified DNA fragments by controlling samples to reach different temperatures so as to achieve the purpose of amplifying the quantity of the DNA fragments by times. Therefore, the accuracy of temperature control and the speed of temperature rise and fall directly affect the efficiency of DNA fragment amplification.
Chinese patent (CN105573368A) discloses a temperature control method of a PCR instrument, which comprises the following steps: the method comprises the following steps that firstly, three temperature sensors are adopted to detect the temperature of each part of the PCR instrument respectively; step two, temperature signal processing; thirdly, the PC calculates a proportional control coefficient, an integral control coefficient and a differential control coefficient by using a genetic algorithm according to the temperature deviation value and the change rate of the deviation, and sends a temperature control command to the main control chip; step four, receiving a control command and adjusting the temperature in real time; and step five, displaying the temperature control curve in real time. Although the patent has simple steps and also uses PID adjusting parameters, the method does not provide a specific algorithm of a PID controller, and the energy consumption is large although the temperature can be adjusted.
Chinese patent (CN108192997A) discloses a temperature control method and a temperature control device for partition of a PCR instrument orifice plate, wherein the method comprises the following steps: acquiring set temperature and current temperature of each partition of the pore plate, wherein the set temperature is the temperature to be controlled and reached by the corresponding partition of the pore plate; obtaining first heating power of each subarea of the pore plate according to the set temperature and the current temperature; and controlling the heating modules of the partitions of the orifice plate to heat according to the first heating power of the partitions of the orifice plate. The first heating power of each subarea of the pore plate is obtained according to the set temperature and the current temperature, and the heating modules of each subarea of the pore plate are controlled by the first heating power corresponding to each subarea to heat the corresponding area.
Disclosure of Invention
The present invention is directed to a method for controlling the temperature of a nucleic acid amplification apparatus, which overcomes the above-mentioned shortcomings of the prior art.
The technical scheme adopted by the invention for solving the technical problem is as follows: in one aspect, a temperature control method for a nucleic acid amplification apparatus is provided, comprising the steps of
S1: inputting a preset temperature T0 into an input device, and respectively acquiring temperature signals of a chip, a primary refrigerating sheet, a secondary refrigerating sheet and a heat dissipation module by a first temperature sensor, a second temperature sensor, a third temperature sensor and a fourth temperature sensor to respectively obtain a first signal temperature, a second signal temperature, a third signal temperature and a fourth signal temperature;
s2: converting the first temperature signal, the second temperature signal, the third temperature signal and the fourth temperature signal acquired in the step S1 into digital quantities T1, T2, T3 and T4 respectively through a temperature signal processing unit, and transmitting the digital quantities to a PID controller for processing;
s3: acquiring a preset temperature T0, acquiring a chip temperature T1 acquired by a first temperature sensor, acquiring temperatures T2 and T3 of a first-stage refrigeration piece and a second-stage refrigeration piece acquired by a second temperature sensor and a third temperature sensor, and acquiring a temperature T4 of a heat dissipation module acquired by a fourth temperature sensor;
s4: averaging the temperatures T2 and T3 of the first refrigerating sheet and the second refrigerating sheet; to obtain T5;
s5: calculating the temperature error e-T1-T0 and the temperature error change rate ec-de/dt;
s6: the fuzzy arithmetic unit takes the temperature error e and the temperature error change rate ec as fuzzy input, and calculates to obtain an output variable proportional coefficient correction value Kp, an integral coefficient correction value Ki and a differential coefficient correction value Kd;
s7: calculating a final proportional correction parameter Kp ' ═ Kp (T1-T5) (T4-T1), a final integral correction parameter Ki ' ═ Ki (T1-T5) (T4-T1), and a final differential correction parameter Kd ' ═ Kd (T1-T5) (T4-T1);
s8: the PID controller controls the execution of the first-stage refrigerating sheet, the second-stage refrigerating sheet and the heat dissipation module according to the final value Kp ' of the proportional coefficient, the final value Ki ' of the integral coefficient and the final value Kd ' of the differential coefficient.
Preferably, step S8 further comprises the steps of k ', Ki ', Kd 'Respectively calculate the proportional output UpKp'. e; integral output Ui＝Ui-1+ Ki' (e-ec); differential output Ud＝Ud-1+ Kd' (e-ea), where ea is the error of the previous cycle; u shaped-1For differential output of the previous time, Ui-1Outputting the integral of the previous time; calculating the output of PID controller as Up+Ui+Ud+ M, M is a correction constant.
Preferably, the step S8 further includes determining a size relationship between output and 0 by the determining unit, and if output is greater than 0, controlling the first-stage refrigeration piece and the second-stage refrigeration piece to continue heating, and controlling and adjusting output powers of the first-stage refrigeration piece and the second-stage refrigeration piece; and if the output is less than 0, controlling the heat dissipation module to continue heat dissipation and controlling and adjusting the output power of the heat dissipation module.
Preferably, the step of controlling and adjusting the output power of the first-stage refrigeration piece and the second-stage refrigeration piece includes comparing the acquired chip temperature T1 acquired by the first temperature sensor with a preset temperature value T0, and if T0-T1>0, increasing the output power of the first-stage refrigeration piece and the second-stage refrigeration piece.
Preferably, the heating determination value is set to Δ T1, the first-stage refrigerant plate and the second-stage refrigerant plate are operated in a 100% power mode if T0-T1> Δ T1, and the output powers of the first-stage refrigerant plate and the second-stage refrigerant plate are operated at α power if 0< T0-T1< Δ T1, and α is T0-T1/Δ T1.
Preferably, the controlling and adjusting the output power of the heat dissipation module includes comparing the acquired chip temperature T1 acquired by the first acquired temperature sensor with a preset temperature value T0, and if T1-T0>0, increasing the output power of the heat dissipation module.
Preferably, the cooling determination value is set to Δ T2, and if T1-T0> Δ T2, the output power of the heat dissipation module is performed in the 100% power mode, and if 0< T1-T0< Δ T2, the output power of the heat dissipation module is performed in the β power mode, and β is T1- Δ T2/T0.
On the other hand, the temperature control system for the nucleic acid amplification instrument comprises an input device, a temperature sensor group, a chip, a primary refrigerating sheet, a secondary refrigerating sheet, a heat dissipation module and a PID control unit; the temperature sensor group comprises a first temperature sensor, a second temperature sensor, a third temperature sensor and a fourth temperature sensor; the PID control unit comprises a temperature signal processing unit, a fuzzy operator and a PID controller, and the fuzzy operator is electrically connected with the PID controller; the temperature signal processing unit is electrically connected with the PID controller; and the PID control unit makes a decision according to the results of the fuzzy arithmetic unit and the PID controller and controls the first-stage refrigerating sheet, the second-stage refrigerating sheet and the heat dissipation module to work.
Preferably, the temperature signal processing unit includes a temperature signal amplifying circuit, a temperature signal filtering circuit, an a/D conversion circuit, and a temperature signal protection circuit; the temperature signal amplifying circuit, the temperature signal filtering circuit, the A/D conversion circuit and the temperature signal protection circuit are sequentially connected with the PID controller; the temperature signal filtering circuit comprises a low-frequency filtering subunit and a high-frequency filtering subunit which are connected in parallel.
Compared with the prior art, the invention has the following beneficial effects:
(1) the fuzzy operation algorithm is added into the traditional PID temperature control algorithm, the corresponding proportional coefficient, integral coefficient and differential coefficient are obtained by carrying out fuzzy operation on the temperature difference and the change rate of the temperature difference, the proportional coefficient, the integral coefficient and the differential coefficient of the temperature control device are further corrected according to the actual condition of the temperature control device, and meanwhile, the decision is made on the first-stage refrigerating sheet, the second-stage refrigerating sheet and the heat dissipation module according to the proportional output, the integral output and the differential output, so that the temperature control accuracy is greatly improved, and the temperature control precision is improved.
(2) The PID control unit provided by the temperature control system for the nucleic acid amplification instrument makes a decision according to the results of the fuzzy operator and the PID controller, and controls the primary refrigerating sheet, the secondary refrigerating sheet and the heat dissipation module to work.
(3) The invention also adjusts the power of the primary refrigeration piece, the secondary refrigeration piece and the heat dissipation module in real time according to the difference value between the preset temperature value and the real-time chip temperature, thereby reducing the energy consumption to the maximum extent, and simultaneously improving the performance of the control system to realize energy conservation and emission reduction.
(4) The PID parameter adjusting method provided by the invention has a simple calculation method, and can scientifically and simply adjust the output power of the primary refrigerating sheet, the secondary refrigerating sheet and the heat dissipation module, so that the temperature control system of the nucleic acid amplification instrument can be well adjusted.
(5) The invention adopts fuzzy PID control, the device has simple structure, good parameter regulation stability and accurate temperature control.
Drawings
FIG. 1 is a flow chart of a method for controlling a nucleic acid amplification apparatus according to the present invention;
Detailed Description
The following detailed description of embodiments of the invention refers to the accompanying drawings.
The invention designs an automatic temperature control system on the basis of the traditional temperature control method, which comprises an input device, a temperature sensor group, a chip, a primary refrigerating sheet, a secondary refrigerating sheet, a heat dissipation module and a PID control unit, wherein the input device is connected with the input device; the temperature sensor group comprises a first temperature sensor, a second temperature sensor, a third temperature sensor and a fourth temperature sensor; the PID control unit comprises a temperature signal processing unit, a fuzzy operator and a PID controller, and the fuzzy operator is electrically connected with the PID controller; the temperature signal processing unit is electrically connected with the PID controller; and the PID control unit makes a decision according to the results of the fuzzy arithmetic unit and the PID controller and controls the first-stage refrigerating sheet, the second-stage refrigerating sheet and the heat dissipation module to work.
The temperature signal processing unit provided by the invention comprises a temperature signal amplifying circuit, a temperature signal filtering circuit, an A/D conversion circuit and a temperature signal protection circuit; the temperature signal amplifying circuit, the temperature signal filtering circuit, the A/D conversion circuit and the temperature signal protection circuit are sequentially connected with the PID controller; the temperature signal filtering circuit comprises a low-frequency filtering subunit and a high-frequency filtering subunit which are connected in parallel.
The first temperature sensor, the second temperature sensor, the third temperature sensor and the fourth temperature sensor provided in the embodiment are all high-sensitivity digital temperature sensors;
the heat dissipation module provided in the present embodiment uses a fan.
As shown in FIG. 1, the present invention also provides a temperature control method for a nucleic acid amplification apparatus, comprising the steps of:
s1: inputting a preset temperature T0 into an input device, and respectively acquiring temperature signals of a chip, a primary refrigerating sheet, a secondary refrigerating sheet and a heat dissipation module by a first temperature sensor, a second temperature sensor, a third temperature sensor and a fourth temperature sensor to respectively obtain a first signal temperature, a second signal temperature, a third signal temperature and a fourth signal temperature;
s2: converting the first temperature signal, the second temperature signal, the third temperature signal and the fourth temperature signal acquired in the step S1 into digital quantities T1, T2, T3 and T4 respectively through a temperature signal processing unit, and transmitting the digital quantities to a PID controller for processing;
s3: acquiring a preset temperature T0, acquiring a chip temperature T1 acquired by a first temperature sensor, acquiring temperatures T2 and T3 of a first-stage refrigeration piece and a second-stage refrigeration piece acquired by a second temperature sensor and a third temperature sensor, and acquiring a temperature T4 of a heat dissipation module acquired by a fourth temperature sensor;
s4: averaging the temperatures T2 and T3 of the first refrigerating sheet and the second refrigerating sheet; to obtain T5;
s5: calculating the temperature error e-T1-T0 and the temperature error change rate ec-de/dt;
s6: the fuzzy arithmetic unit takes the temperature error e and the temperature error change rate ec as fuzzy input, and calculates to obtain an output variable proportional coefficient correction value Kp, an integral coefficient correction value Ki and a differential coefficient correction value Kd;
s7: calculating a final proportional correction parameter Kp ' ═ Kp (T1-T5) (T4-T1), a final integral correction parameter Ki ' ═ Ki (T1-T5) (T4-T1), and a final differential correction parameter Kd ' ═ Kd (T1-T5) (T4-T1);
s8: the PID controller controls the execution of the first-stage refrigerating sheet, the second-stage refrigerating sheet and the heat dissipation module according to the final value Kp ' of the proportional coefficient, the final value Ki ' of the integral coefficient and the final value Kd ' of the differential coefficient.
Wherein, step S8 further comprises calculating the proportional output U according to Kp ', Ki' and KdpKp'. e; integral output Ui＝Ui-1+ Ki' (e-ec); differential output Ud＝Ud-1+ Kd' (e-ea), where ea is the error of the previous cycle; u shaped-1For differential output of the previous time, Ui-1Outputting the integral of the previous time; calculating the output of PID controller as Up+Ui+Ud+ M, M is a correction constant.
Step S8 also includes judging the size relationship between output and 0 through the judging unit, if output is greater than 0, controlling the first-stage refrigeration piece and the second-stage refrigeration piece to continue heating, and controlling and adjusting the output power of the first-stage refrigeration piece and the second-stage refrigeration piece; and if the output is less than 0, controlling the heat dissipation module to continue heat dissipation and controlling and adjusting the output power of the heat dissipation module.
The step of controlling and adjusting the output power of the first-stage refrigerating piece and the second-stage refrigerating piece comprises the step of comparing the chip temperature T1 acquired by the acquired first temperature sensor with a preset temperature value T0, and if the T0-T1 is greater than 0, the output power of the first-stage refrigerating piece and the output power of the second-stage refrigerating piece are increased.
And setting the heating judgment value to be delta T1, if T0-T1 is greater than delta T1, operating the first-stage refrigerating sheet and the second-stage refrigerating sheet in a 100% power mode, and if 0< T0-T1< delta T1, operating the output power of the first-stage refrigerating sheet and the second-stage refrigerating sheet in α power, wherein α is T0-T1/delta T1.
The step of controlling and adjusting the output power of the heat dissipation module comprises the step of comparing the acquired chip temperature T1 acquired by the first acquired temperature sensor with a preset temperature value T0, and if T1-T0>0, the output power of the heat dissipation module is increased.
The cooling determination value is set to Δ T2, and if T1-T0> Δ T2, the output power of the heat dissipation module is performed in the 100% power mode, and if 0< T1-T0< Δ T2, the output power of the heat dissipation module is performed in the β power mode, and β is T1- Δ T2/T0.
The fuzzy operation algorithm is added into the traditional PID temperature control algorithm, the corresponding proportional coefficient, integral coefficient and differential coefficient are obtained by carrying out fuzzy operation on the temperature difference and the change rate of the temperature difference, the proportional coefficient, the integral coefficient and the differential coefficient of the temperature control device are further corrected according to the actual condition of the temperature control device, and meanwhile, the decision is made on the first-stage refrigerating sheet, the second-stage refrigerating sheet and the heat dissipation module according to the proportional output, the integral output and the differential output, so that the temperature control accuracy is greatly improved, and the temperature control precision is improved. Meanwhile, the PID parameter adjusting method provided by the invention has good parameter adjusting stability and simple calculation method, and can scientifically adjust the output power of the primary refrigerating sheet, the secondary refrigerating sheet and the heat dissipation module, thereby realizing good adjustment of the temperature control system of the nucleic acid amplification instrument. And the energy consumption can be reduced to the maximum extent.
Although the preferred embodiments of the present invention have been described in detail with reference to the accompanying drawings, the present invention is not limited to the above embodiments, and various changes can be made within the knowledge of those skilled in the art without departing from the spirit of the present invention.
Many other changes and modifications can be made without departing from the spirit and scope of the invention. It is to be understood that the invention is not to be limited to the specific embodiments, but only by the scope of the appended claims.
Claims (5)
1. A temperature control method for a nucleic acid amplification instrument, comprising the steps of:
s1: inputting a preset temperature T0 into an input device, and respectively acquiring temperature signals of a chip, a primary refrigerating sheet, a secondary refrigerating sheet and a heat dissipation module by a first temperature sensor, a second temperature sensor, a third temperature sensor and a fourth temperature sensor to respectively obtain a first signal temperature, a second signal temperature, a third signal temperature and a fourth signal temperature;
s2: converting the first temperature signal, the second temperature signal, the third temperature signal and the fourth temperature signal acquired in the step S1 into digital quantities T1, T2, T3 and T4 respectively through a temperature signal processing unit, and transmitting the digital quantities to a PID controller for processing;
s3: acquiring a preset temperature T0, acquiring a chip temperature T1 acquired by a first temperature sensor, acquiring temperatures T2 and T3 of a first-stage refrigeration piece and a second-stage refrigeration piece acquired by a second temperature sensor and a third temperature sensor, and acquiring a temperature T4 of a heat dissipation module acquired by a fourth temperature sensor;
s4: averaging the temperatures T2 and T3 of the first refrigerating sheet and the second refrigerating sheet; to obtain T5;
s5: calculating the temperature error e-T1-T0 and the temperature error change rate ec-de/dt;
s6: the fuzzy arithmetic unit takes the temperature error e and the temperature error change rate ec as fuzzy input, and calculates to obtain an output variable proportional coefficient correction value Kp, an integral coefficient correction value Ki and a differential coefficient correction value Kd;
s7: calculating a final proportional correction parameter Kp ' ═ Kp (T1-T5) (T4-T1), a final integral correction parameter Ki ' ═ Ki (T1-T5) (T4-T1), and a final differential correction parameter Kd ' ═ Kd (T1-T5) (T4-T1);
s8: the PID controller controls the execution of the primary refrigeration piece, the secondary refrigeration piece and the heat dissipation module according to the final value Kp ' of the proportional coefficient, the final value Ki ' of the integral coefficient and the final value Kd ' of the differential coefficient;
step S8 further includes calculating proportional outputs Up to Kp 'e according to Kp', Ki ', KD'; integral output Ui＝Ui-1+ Ki' (e-ec); differential output Ud＝Ud-1+ Kd' (e-ea), where ea is the error of the previous cycle; calculating the output of PID controller as Up+Ui+Ud+ M, M is a correction constant;
step S8 also includes judging the size relationship between output and 0 through the judging unit, if output is greater than 0, controlling the first-stage refrigeration piece and the second-stage refrigeration piece to continue heating, and controlling and adjusting the output power of the first-stage refrigeration piece and the second-stage refrigeration piece; if output is less than 0, controlling the heat dissipation module to continue heat dissipation, and controlling and adjusting the output power of the heat dissipation module;
controlling and adjusting the output power of the first-stage refrigerating piece and the second-stage refrigerating piece comprises comparing the acquired chip temperature T1 acquired by the first temperature sensor with a preset temperature value T0, and if the T0-T1 is greater than 0, increasing the output power of the first-stage refrigerating piece and the second-stage refrigerating piece;
and setting the heating judgment value to be delta T1, if T0-T1 is greater than delta T1, operating the first-stage refrigerating sheet and the second-stage refrigerating sheet in a 100% power mode, and if 0< T0-T1< delta T1, operating the output power of the first-stage refrigerating sheet and the second-stage refrigerating sheet in α power, wherein α is T0-T1/delta T1.
2. The temperature control method for a nucleic acid amplification apparatus according to claim 1, wherein the controlling and adjusting the output power of the heat sink module comprises comparing the obtained chip temperature T1 collected by the first temperature sensor with a preset temperature value T0, and if T1-T0>0, increasing the output power of the heat sink module.
3. The temperature control method for a nucleic acid amplification apparatus according to claim 2, wherein the cooling determination value is set to Δ T2, and when T1-T0> Δ T2, the output power of the heat dissipation module is performed in a 100% power mode, and when 0< T1-T0< Δ T2, the output power of the heat dissipation module is performed in a β power mode, and β is T1- Δ T2/T0.
4. The control system for the temperature control method of the nucleic acid amplification instrument according to any one of claims 1 to 3, comprising an input device, a temperature sensor group, a chip, a primary refrigeration piece, a secondary refrigeration piece, a heat dissipation module, and a PID control unit; the temperature sensor group comprises a first temperature sensor, a second temperature sensor, a third temperature sensor and a fourth temperature sensor; the PID control unit comprises a temperature signal processing unit, a fuzzy operator and a PID controller, and the fuzzy operator is electrically connected with the PID controller; the temperature signal processing unit is electrically connected with the PID controller; and the PID control unit makes a decision according to the results of the fuzzy arithmetic unit and the PID controller and controls the first-stage refrigerating sheet, the second-stage refrigerating sheet and the heat dissipation module to work.
5. The control system for the temperature control method of a nucleic acid amplification apparatus according to claim 4, wherein the temperature signal processing unit comprises a temperature signal amplifying circuit, a temperature signal filtering circuit, an A/D converting circuit, and a temperature signal protecting circuit; the temperature signal amplifying circuit, the temperature signal filtering circuit, the A/D conversion circuit and the temperature signal protection circuit are sequentially connected with the PID controller; the temperature signal filtering circuit comprises a low-frequency filtering subunit and a high-frequency filtering subunit which are connected in parallel.
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| CN110684643B (en) * | 2019-10-30 | 2021-03-19 | 宁波胤瑞生物医学仪器有限责任公司 | Temperature conduction system for nucleic acid amplification instrument |
| CN113377140B (en) * | 2021-06-09 | 2022-10-04 | 厦门大学 | Temperature control method and device for nucleic acid detection device |
| CN113485478A (en) * | 2021-07-07 | 2021-10-08 | 维柯基科技(上海)有限公司 | Temperature control method, device and system of PCR amplification instrument, computer equipment and computer readable storage medium |
| US11938485B2 (en) | 2021-12-07 | 2024-03-26 | Industrial Technology Research Institute | Heating device for convective polymerase chain reaction |
| CN114812005B (en) * | 2022-06-28 | 2022-10-11 | 合肥中科国腾生物科技有限公司 | Active rapid heat dissipation method and device of thermal cycler based on thermoelectric refrigeration piece |
| CN115197835B (en) * | 2022-09-15 | 2023-07-11 | 浙江正合谷生物科技有限公司 | Nucleic acid amplification fluorescence quantitative temperature control system based on self-learning |
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| CN104808708A (en) * | 2015-04-22 | 2015-07-29 | 重庆工商职业学院 | Method and system for self-adjusting fuzzy PID (Proportion Integration Differentiation) parameters in furnace temperature control system |
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