A PCR instrument self-optimization control method and system

Abstract

The present invention discloses a self-optimization control method and system for a PCR instrument, and relates to the technical field of data processing and control of a PCR instrument, including determining reaction stage target information and a control cycle corresponding to each reaction stage according to auxiliary reaction information and information of samples to be processed, thereby configuring the reaction stage target information of the event, thereby ensuring the accuracy of monitoring the reaction stage. Based on real-time reaction data and real-time auxiliary reaction data, the deviation value and the comprehensive influence coefficient of the reaction system of each sub-reaction stage are generated, the fluctuation situation is described by the deviation value and the comprehensive influence coefficient of the reaction system, and the temperature rate of the PCR instrument of the next sub-reaction stage is adjusted, thereby improving the control effect and adaptability within a single stage. The temperature rate of the PCR instrument of the next reaction stage is controlled according to the reaction stage evaluation index and type of the previous sub-reaction stage, and the temperature rate of the PCR instrument of the next reaction stage is adjusted, thereby improving the control accuracy and adaptability between multiple stages.

Description

Technical Field

[0001] The present invention relates to the technical field of data processing and control of PCR instruments, and in particular to a self-optimizing control method and system for a PCR instrument. Background Art

[0002] Traditional PCR instruments rely on manually set temperature and time parameters, but due to changes in experimental conditions and differences in operator experience, experimental results are often unstable and have poor repeatability. With the deepening of molecular biology research and the increase in clinical diagnostic needs, higher requirements are placed on the automation and intelligent control of PCR instruments. The self-optimizing control technology of PCR instruments can automatically adjust reaction conditions to adapt to different experimental needs by integrating advanced temperature control algorithms, real-time monitoring systems and data analysis software. This technology can monitor key parameters such as temperature and fluorescence signals during the PCR reaction in real time, and analyze these data through algorithms to automatically adjust key parameters such as temperature rate and annealing temperature to optimize the PCR reaction process.

[0003] In the prior art, the self-optimizing control of the PCR instrument often only sets a fixed temperature rate to cool down or heat up, and does not take into account the actual complex experimental requirements, resulting in poor control effect, low adaptability and poor accuracy of the PCR instrument.

[0004] Therefore, how to improve the effect, adaptability and accuracy of PCR instrument control is a technical problem that needs to be solved. Summary of the invention

[0005] The purpose of the present invention is to solve the problems of poor control effect, low adaptability and poor accuracy of PCR instrument in the prior art, and to propose a self-optimizing control method for PCR instrument, which includes:

[0006] Obtaining information of samples to be processed and basic information of the PCR instrument, configuring auxiliary reaction information according to the information of samples to be processed and the basic information of the PCR instrument, and determining reaction stage target information and a control cycle corresponding to each reaction stage according to the auxiliary reaction information and the information of samples to be processed;

[0007] Each reaction stage is divided into multiple sub-reaction stages according to the control cycle, and the reaction stage target information is respectively configured on each sub-reaction stage, and the real-time reaction data and real-time auxiliary reaction data of each sub-reaction stage on the PCR instrument are monitored, and the deviation value and the comprehensive influence coefficient of the reaction system of each sub-reaction stage are generated based on the real-time reaction data and the real-time auxiliary reaction data;

[0008] The temperature rate of the PCR instrument in the next sub-reaction stage is controlled by combining the deviation value of the previous sub-reaction stage and the comprehensive coefficient of the reaction system, so as to complete the optimization control of a single reaction stage;

[0009] After each reaction stage is completed, the reaction stage evaluation index and evaluation index type are defined according to the overall reaction data of the reaction stage, and the temperature rate of the PCR instrument of the next reaction stage is controlled according to the reaction stage evaluation index and type of the previous sub-reaction stage, so as to complete the continuous optimization control between multiple reaction stages;

[0010] Among them, the reaction stages involved in the PCR instrument include denaturation stage, annealing stage and extension stage.

[0011] In some embodiments of the present application, the reaction stage target information and the control period corresponding to each reaction stage are determined according to the auxiliary reaction information and the sample information to be processed, including:

[0012] The target information of the reaction stage includes the target time and target temperature, and the auxiliary reaction information and the information of the sample to be processed include time-related information, temperature-related information and other information;

[0013] Find similar reaction records in the historical reaction records through the information of the sample to be processed and the auxiliary reaction information, record them as similar reaction records, and determine the first target time range and the first target temperature range corresponding to the denaturation stage, the annealing stage and the extension stage respectively according to these similar reaction records;

[0014] Comprehensively determine the second target time range and the second target temperature range corresponding to the denaturation stage, the annealing stage, and the extension stage, respectively, according to the time-related information and the temperature-related information in the auxiliary reaction information and the information of the sample to be processed;

[0015] The intersection range of the first target duration range and the second target duration range is used as the target duration of each reaction stage, and the intersection range of the first target temperature range and the second target temperature range is used as the target temperature of each reaction stage;

[0016] The control cycles corresponding to the denaturation phase, the annealing phase and the extension phase are set according to the auxiliary reaction information, the sample to be processed information and the reaction phase target information.

[0017] In some embodiments of the present application, the control cycles corresponding to the denaturation phase, the annealing phase, and the extension phase are set according to the auxiliary reaction information, the sample information to be processed, and the reaction phase target information, including:

[0018] In similar reaction records, each reaction stability index in the auxiliary reaction information and the sample to be processed information is evaluated, each reaction stability index is normalized, and a target parameter change curve of the target parameter of each reaction stage in the denaturation stage, the annealing stage, and the extension stage is constructed according to the similar reaction records, the target parameter change curve is split into a plurality of time-continuous partial curves, the average value of the slope change on each partial curve is calculated, and the average value of the slope change on each partial curve is combined to determine the average value of the slope change in the reaction stage;

[0019] The stability level corresponding to the denaturation stage, annealing stage and extension stage is determined according to the target time, the average value of the slope change of the reaction stage and the reaction stability index;

[0020] ;

[0021] in, is the stability level, is the number of reaction stability indicators in the auxiliary reaction information and the sample information to be processed, The auxiliary reaction information and the sample information to be processed The weight of each reaction stability index, The auxiliary reaction information and the sample information to be processed The size of the reaction stability index, is the average value of the slope change during the reaction phase, For stable duration, It represents the stable duration mapped from the average value of the slope change in the reaction phase. is the median of the target duration, are the conversion coefficients of the stable duration and the target duration, respectively. are the first constant and the second constant respectively, [] is the rounding symbol;

[0022] The control period corresponding to each reaction stage is set by the stability level, and different stability levels correspond to different control period sizes.

[0023] In some embodiments of the present application, each reaction stage is divided into multiple sub-reaction stages according to the control cycle, and the reaction stage target information is respectively configured on each sub-reaction stage, including:

[0024] The corresponding reaction stage is divided into multiple sub-reaction stages according to the size of the control cycle, and the multiple sub-reaction stages are continuous in time. The sub-target temperature and sub-target fluorescence signal amount corresponding to each sub-reaction stage are set according to the target duration and target temperature of each reaction stage.

[0025] In some embodiments of the present application, the deviation value of each sub-reaction stage and the comprehensive influence coefficient of the reaction system are generated based on the real-time reaction data and the real-time auxiliary reaction data, including:

[0026] Compare the deviations of the real-time reaction data in each sub-reaction stage with the sub-target temperature and the sub-target fluorescence signal, generate a first deviation and a second deviation, and determine the deviation value of each sub-reaction stage by combining the first deviation and the second deviation;

[0027] The real-time reaction data and the real-time auxiliary reaction data are input into the preset reaction system evaluation model to obtain the comprehensive influence coefficient of the reaction system.

[0028] In some embodiments of the present application, the temperature rate of the PCR instrument in the next sub-reaction stage is controlled by combining the deviation value of the previous sub-reaction stage and the comprehensive coefficient of the reaction system, including:

[0029] Detect whether there is non-specific amplification in the previous sub-reaction stage. If there is non-specific amplification in the previous sub-reaction stage, identify the properties of the non-specific amplification, and control the temperature rate of the PCR instrument in the next sub-reaction stage according to the properties of the non-specific amplification, the deviation value and the comprehensive coefficient of the reaction system;

[0030] ;

[0031] in, For the The target value of the PCR instrument temperature rate for each sub-reaction stage, For the The initial value of the PCR instrument temperature rate for each sub-reaction stage, For the The comprehensive coefficient of the reaction system in each sub-reaction stage is It is The deviation value of each sub-reaction stage, are the conversion factor of the bias value and nonspecific amplification, is the number of attribute features of nonspecific amplification, For the The weight of the attribute features, For the The non-specific amplification of the sub-reaction stage The size of the attribute feature, is the third constant.

[0032] In some embodiments of the present application, after each reaction stage is completed, the reaction stage evaluation index and the evaluation index type are defined according to the overall reaction data of the reaction stage, including:

[0033] Determine the reaction stage evaluation index based on the temperature and fluorescence signal in the overall reaction data of the reaction stage;

[0034] Evaluation indicator types include non-cyclic evaluation indicators and cyclic evaluation indicators;

[0035] The reaction stage evaluation indexes of the denaturation stage and the annealing stage are defined as non-cyclic evaluation indexes, and the evaluation index of the extension stage is defined as cyclic evaluation index.

[0036] In some embodiments of the present application, the temperature rate of the PCR instrument in the next reaction stage is controlled according to the reaction stage evaluation index and type of the previous sub-reaction stage, including:

[0037] When the previous sub-reaction stage is a denaturation stage or an annealing stage, the temperature rate of the PCR instrument in the next reaction stage is directly controlled according to the non-circulation evaluation index corresponding to each stage;

[0038] When the previous sub-reaction stage is the extension stage, the non-cyclic evaluation index and the cyclic evaluation index are integrated to obtain a comprehensive evaluation index, and the temperature rate of the PCR instrument in the next reaction stage is controlled by the comprehensive evaluation index.

[0039] Correspondingly, the present application also provides a PCR instrument self-optimization control system, comprising:

[0040] A setting module is used to obtain the information of the sample to be processed and the basic information of the PCR instrument, configure the auxiliary reaction information according to the information of the sample to be processed and the basic information of the PCR instrument, and determine the reaction stage target information and the control cycle corresponding to each reaction stage according to the auxiliary reaction information and the information of the sample to be processed;

[0041] An analysis module is used to split each reaction stage into multiple sub-reaction stages according to the control cycle, configure the reaction stage target information on each sub-reaction stage, monitor the real-time reaction data and real-time auxiliary reaction data of each sub-reaction stage on the PCR instrument, and generate a deviation value and a comprehensive influence coefficient of the reaction system for each sub-reaction stage based on the real-time reaction data and the real-time auxiliary reaction data;

[0042] The first optimization module is used to control the temperature rate of the PCR instrument in the next sub-reaction stage by combining the deviation value of the previous sub-reaction stage and the comprehensive coefficient of the reaction system, so as to complete the optimization control of a single reaction stage;

[0043] The second optimization module is used to define the reaction stage evaluation index and the evaluation index type according to the overall reaction data of the reaction stage after each reaction stage is completed, and control the temperature rate of the PCR instrument of the next reaction stage according to the reaction stage evaluation index and type of the previous sub-reaction stage, so as to complete the continuous optimization control between multiple reaction stages;

[0044] Among them, the reaction stages involved in the PCR instrument include denaturation stage, annealing stage and extension stage.

[0045] Compared with the prior art, the present invention has the following beneficial effects:

[0046] 1. Determine the reaction stage target information and the control cycle corresponding to each reaction stage based on the auxiliary reaction information and the information of the sample to be processed, so as to configure the reaction stage target information to ensure the accuracy of the reaction stage monitoring, and set the control cycle to carry out detailed control of the subsequent stages.

[0047] 2. Generate the deviation value and comprehensive influence coefficient of the reaction system for each sub-reaction stage based on real-time reaction data and real-time auxiliary reaction data. Use the deviation value and comprehensive influence coefficient of the reaction system to describe the fluctuation situation, and adjust the temperature rate of the PCR instrument in the next sub-reaction stage to improve the control effect and adaptability within a single stage.

[0048] 3. Control the temperature rate of the PCR instrument in the next reaction stage according to the reaction stage evaluation index and type of the previous sub-reaction stage. Describe the reaction effect of each reaction stage through the reaction stage evaluation index and type, and adjust the temperature rate of the PCR instrument in the next reaction stage to improve the control accuracy and adaptability between multiple stages. BRIEF DESCRIPTION OF THE DRAWINGS

[0049] Figure 1 A schematic diagram of a process flow of a PCR instrument self-optimization control method proposed by the present invention;

[0050] Figure 2 This is a schematic diagram of the structure of a self-optimizing control system for a PCR instrument proposed by the present invention. DETAILED DESCRIPTION

[0051] The technical solutions in the embodiments of the present invention will be described clearly and completely below in conjunction with the drawings in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, rather than all the embodiments.

[0052] Reference Figure 1 , a PCR instrument self-optimization control method, comprising the following steps:

[0053] Step S101, obtain the sample information to be processed and the basic information of the PCR instrument, configure the auxiliary reaction information according to the sample information to be processed and the basic information of the PCR instrument, and determine the reaction stage target information and the control cycle corresponding to each reaction stage according to the auxiliary reaction information and the sample information to be processed.

[0054] In this embodiment, the information of the sample to be processed: the type, volume, concentration, etc. of the sample are obtained. Basic information of the PCR instrument: the model of the PCR instrument, the maximum / minimum temperature range, the heating / cooling rate, the settable reaction time and other parameters are obtained. Auxiliary reaction information includes buffer type, enzyme type and amount, primer concentration, etc. The reaction stages involved in the PCR instrument include denaturation stage, annealing stage and extension stage. A complete process of denaturation stage-annealing stage-extension stage is a single cycle in sequence, and dozens of such cycles will be performed later. The target information of the reaction stage is the target temperature, time, etc. of each stage. For example, denaturation stage: the target temperature is 94-98°C, and the duration is determined according to the characteristics of the enzyme, usually 30 seconds to 1 minute. Annealing stage: the target temperature is set according to the Tm value of the primer, usually between 50-65°C, and the duration is about 30 seconds to 1 minute. Extension stage: the target temperature is 72°C (for most Taq enzymes), and the duration is determined according to the fragment length and enzyme efficiency, usually 1-2 minutes.

[0055] In some embodiments of the present application, the reaction stage target information and the control period corresponding to each reaction stage are determined according to the auxiliary reaction information and the sample information to be processed, including:

[0056] The target information of the reaction stage includes the target time and target temperature, and the auxiliary reaction information and the information of the sample to be processed include time-related information, temperature-related information and other information;

[0057] Find similar reaction records in the historical reaction records through the information of the sample to be processed and the auxiliary reaction information, record them as similar reaction records, and determine the first target time range and the first target temperature range corresponding to the denaturation stage, the annealing stage and the extension stage respectively according to these similar reaction records;

[0058] Comprehensively determine the second target time range and the second target temperature range corresponding to the denaturation stage, the annealing stage, and the extension stage, respectively, according to the time-related information and the temperature-related information in the auxiliary reaction information and the information of the sample to be processed;

[0059] The intersection range of the first target duration range and the second target duration range is used as the target duration of each reaction stage, and the intersection range of the first target temperature range and the second target temperature range is used as the target temperature of each reaction stage;

[0060] The control cycles corresponding to the denaturation phase, the annealing phase and the extension phase are set according to the auxiliary reaction information, the sample to be processed information and the reaction phase target information.

[0061] In this embodiment, similar reaction records are searched in the past historical reaction records, for example, records with the same sample and similar auxiliary reaction information, and the first target time range and the first target temperature range are screened out from the similar reaction records.

[0062] In this embodiment, the auxiliary reaction information and the information of the sample to be processed include time-related information, temperature-related information and other information. Time-related information refers to information that affects the duration of the reaction phase, such as the concentration and purity of the template DNA (a high concentration of template DNA may shorten the time required for the extension phase, while a low purity may affect the reaction efficiency), the length of the target fragment (the amplification of a longer fragment requires a longer extension time), etc. Temperature-related information refers to information that affects the target temperature of the reaction phase, such as enzyme properties (understanding the optimal temperature range of the DNA polymerase used (such as Taq enzyme) and its tolerance to high and low temperatures), the Tm value of the primer (determine the target temperature of the annealing phase based on the melting temperature (Tm value) of the primer. The Tm value is usually related to the length, sequence and GC content of the primer) and the composition of the reaction system (considering the concentration and properties of components such as buffer, dNTPs (deoxyribonucleoside triphosphates), and Mg²⁺ ions in the reaction system, which may affect the activity and reaction rate of the enzyme), etc.

[0063] In this embodiment, the second target duration range and the second target temperature range are determined by comprehensively considering the duration-related information and the temperature-related information, and the target duration and the target temperature are determined by combining the first target duration range, the first target temperature range, the second target duration range and the second target temperature range.

[0064] In some embodiments of the present application, the control cycles corresponding to the denaturation phase, the annealing phase, and the extension phase are set according to the auxiliary reaction information, the sample information to be processed, and the reaction phase target information, including:

[0065] In similar reaction records, each reaction stability index in the auxiliary reaction information and the sample to be processed information is evaluated, each reaction stability index is normalized, and a target parameter change curve of the target parameter of each reaction stage in the denaturation stage, the annealing stage, and the extension stage is constructed according to the similar reaction records, the target parameter change curve is split into a plurality of time-continuous partial curves, the average value of the slope change on each partial curve is calculated, and the average value of the slope change on each partial curve is combined to determine the average value of the slope change in the reaction stage;

[0066] The stability level corresponding to the denaturation stage, annealing stage and extension stage is determined according to the target time, the average value of the slope change of the reaction stage and the reaction stability index;

[0067] ;

[0068] in, is the stability level, is the number of reaction stability indicators in the auxiliary reaction information and the sample information to be processed, The auxiliary reaction information and the sample information to be processed The weight of each reaction stability index, The auxiliary reaction information and the sample information to be processed The size of the reaction stability index, is the average value of the slope change during the reaction phase, For stable duration, It represents the stable duration mapped from the average value of the slope change in the reaction phase. is the median of the target duration, are the conversion coefficients of the stable duration and the target duration, respectively. are the first constant and the second constant respectively, [] is the rounding symbol;

[0069] The control period corresponding to each reaction stage is set by the stability level, and different stability levels correspond to different control period sizes.

[0070] In this embodiment, each reaction stability index in the auxiliary reaction information and the sample information to be processed is evaluated in the similar reaction record. The stability index refers to an index that affects the change of the system, such as enzyme activity, primer quality, sample concentration, etc. These conditions will affect the change of the overall reaction.

[0071] In this embodiment, the target parameters in the reaction stage may not be directly obtained or monitored, and other parameters are needed to indirectly reflect them. The target parameters of each reaction stage in the denaturation stage, annealing stage and extension stage are as follows:

[0072] Transformation stage

[0073] Target parameter: DNA single-strand degree

[0074] Definition: Describes the degree of completion of the conversion of a DNA template from a double-stranded state to a single-stranded state.

[0075] Evaluation method: Real-time evaluation is performed by monitoring temperature changes and the unwinding of DNA chains. When the DNA chains are observed to be completely unwound and the temperature reaches the preset denaturation temperature (target temperature), it can be considered that the degree of DNA single-stranding has reached the highest level.

[0076] Annealing stage

[0077] Target parameter: Primer binding efficiency

[0078] Definition: Describes the ratio or degree of correct binding of primers to template DNA.

[0079] Evaluation method: Real-time evaluation is performed by monitoring the changes in fluorescence signals. When the fluorescence signal reaches the preset threshold, the primer binding efficiency is considered to be optimal.

[0080] Extension Phase

[0081] Target parameter: rate of new DNA chain synthesis

[0082] Definition: Describes the rate at which new DNA chains are synthesized during the elongation phase.

[0083] Evaluation method: Real-time evaluation is performed by monitoring the growth rate of the fluorescence signal. When the growth rate of the fluorescence signal reaches a stable and high level, it can be considered that the synthesis rate of the new DNA chain has reached the optimal level.

[0084] In this embodiment, the average value of the slope change in the reaction stage is determined by combining the average value of the slope change on each partial curve, and the change of the reaction in this stage is described by the average value of the slope change. It means that the stability level is determined by correcting the sum of the stability indicators by the fluctuation duration and the target duration.

[0085] It can be understood that the control cycle length determined here is less than the target duration.

[0086] Step S102, split each reaction stage into multiple sub-reaction stages according to the control cycle, configure the reaction stage target information on each sub-reaction stage respectively, monitor the real-time reaction data and real-time auxiliary reaction data of each sub-reaction stage on the PCR instrument, and generate the deviation value and the comprehensive influence coefficient of the reaction system of each sub-reaction stage based on the real-time reaction data and real-time auxiliary reaction data.

[0087] In this embodiment, each reaction stage is split, and the fluctuation of the sub-reaction stage is described by the deviation value and the comprehensive influence coefficient of the reaction system.

[0088] In some embodiments of the present application, each reaction stage is divided into multiple sub-reaction stages according to the control cycle, and the reaction stage target information is respectively configured on each sub-reaction stage, including:

[0089] The corresponding reaction stage is divided into multiple sub-reaction stages according to the size of the control cycle, and the multiple sub-reaction stages are continuous in time. The sub-target temperature and sub-target fluorescence signal amount corresponding to each sub-reaction stage are set according to the target duration and target temperature of each reaction stage.

[0090] In this embodiment, the sub-target temperature and sub-target fluorescence signal amount (determined according to the duration) of each sub-reaction stage are set according to the target duration and target temperature of the overall reaction stage. The fluorescence signal is usually used to monitor the progress of the reaction or the formation of the product. By setting the target fluorescence signal amount, the reaction process can be monitored in real time.

[0091] In some embodiments of the present application, the deviation value of each sub-reaction stage and the comprehensive influence coefficient of the reaction system are generated based on the real-time reaction data and the real-time auxiliary reaction data, including:

[0092] Compare the deviations of the real-time reaction data in each sub-reaction stage with the sub-target temperature and the sub-target fluorescence signal, generate a first deviation and a second deviation, and determine the deviation value of each sub-reaction stage by combining the first deviation and the second deviation;

[0093] The real-time reaction data and the real-time auxiliary reaction data are input into the preset reaction system evaluation model to obtain the comprehensive influence coefficient of the reaction system.

[0094] In this embodiment, the preset reaction system evaluation model may be a mechanism model based on physical and chemical principles, which reflects the stability and efficiency of the reaction system and mainly integrates real-time auxiliary reaction data to determine a comprehensive influence coefficient of the reaction system.

[0095] Step S103, combining the deviation value of the previous sub-reaction stage and the comprehensive coefficient of the reaction system to control the temperature rate of the PCR instrument in the next sub-reaction stage, so as to complete the optimization control of a single reaction stage.

[0096] In this embodiment, by controlling the temperature rate (speed of heating and cooling) of the PCR instrument, for a single reaction stage, the temperature rate of the PCR instrument in the next reaction stage is adjusted according to the deviation value of the previous sub-reaction stage and the comprehensive coefficient of the reaction system.

[0097] In some embodiments of the present application, the temperature rate of the PCR instrument in the next sub-reaction stage is controlled by combining the deviation value of the previous sub-reaction stage and the comprehensive coefficient of the reaction system, including:

[0098] Detect whether there is non-specific amplification in the previous sub-reaction stage. If there is non-specific amplification in the previous sub-reaction stage, identify the properties of the non-specific amplification, and control the temperature rate of the PCR instrument in the next sub-reaction stage according to the properties of the non-specific amplification, the deviation value and the comprehensive coefficient of the reaction system;

[0099] ;

[0100] in, For the The target value of the PCR instrument temperature rate for each sub-reaction stage, For the The initial value of the PCR instrument temperature rate for each sub-reaction stage, For the The comprehensive coefficient of the reaction system in each sub-reaction stage is It is The deviation value of each sub-reaction stage, are the conversion factor of the bias value and nonspecific amplification, is the number of attribute features of nonspecific amplification, For the The weight of the attribute features, For the The non-specific amplification of the sub-reaction stage The size of the attribute feature, is the third constant.

[0101] In this example, nonspecific amplification is a common problem during the PCR reaction, which may result in the appearance of amplified bands that are inconsistent with the target band size in the experimental results. The degree of nonspecific amplification can be evaluated from the following attribute characteristics:

[0102] Number of amplified bands: Nonspecific amplification usually results in multiple amplified bands rather than a single target band. The more amplified bands there are, the more serious the nonspecific amplification may be.

[0103] Brightness of amplification bands: The brightness of bands produced by nonspecific amplification can also reflect its extent. Brighter bands indicate a larger amount of nonspecific amplification products, which may indicate a higher degree of nonspecific amplification.

[0104] Growth rate: The growth rate of nonspecific amplification products is also an important evaluation indicator. If the amount of nonspecific amplification products increases rapidly in a short period of time, it means that the degree of nonspecific amplification may be serious.

[0105] In this embodiment, It indicates the correction of the initial value of the PCR instrument temperature rate by the deviation value and non-specific amplification.

[0106] Step S104, after each reaction stage is completed, the reaction stage evaluation index and the evaluation index type are defined according to the overall reaction data of the reaction stage, and the temperature rate of the PCR instrument of the next reaction stage is controlled according to the reaction stage evaluation index and the type of the previous sub-reaction stage, so as to complete the continuous optimization control between multiple reaction stages;

[0107] Among them, the reaction stages involved in the PCR instrument include denaturation stage, annealing stage and extension stage.

[0108] In some embodiments of the present application, after each reaction stage is completed, the reaction stage evaluation index and the evaluation index type are defined according to the overall reaction data of the reaction stage, including:

[0109] Determine the reaction stage evaluation index based on the temperature and fluorescence signal in the overall reaction data of the reaction stage;

[0110] Evaluation indicator types include non-cyclic evaluation indicators and cyclic evaluation indicators;

[0111] The reaction stage evaluation indexes of the denaturation stage and the annealing stage are defined as non-cyclic evaluation indexes, and the evaluation index of the extension stage is defined as cyclic evaluation index.

[0112] In this embodiment, the reaction stage evaluation index is determined based on the temperature and fluorescence signal in the overall reaction data of the reaction stage. For the evaluation index, for the denaturation stage, it is the time when the temperature reaches the preset denaturation temperature; for the annealing stage, it is the time when the fluorescence signal reaches the preset threshold value; for the extension stage, it is the time when the fluorescence signal growth rate reaches the preset threshold value.

[0113] It can be understood that the above-mentioned preset threshold is the target temperature, etc., and the evaluation index is obtained by combining the target duration and the time when the time reaches the preset threshold.

[0114] In this embodiment, the non-cyclic evaluation index means that the denaturation stage and annealing stage adjust the control of the next stage within a single cycle. The cyclic evaluation index is the extension stage that adjusts the denaturation stage within the next cycle, involving the next cycle.

[0115] In some embodiments of the present application, the temperature rate of the PCR instrument in the next reaction stage is controlled according to the reaction stage evaluation index and type of the previous sub-reaction stage, including:

[0116] When the previous sub-reaction stage is a denaturation stage or an annealing stage, the temperature rate of the PCR instrument in the next reaction stage is directly controlled according to the non-circulation evaluation index corresponding to each stage;

[0117] When the previous sub-reaction stage is the extension stage, the non-cyclic evaluation index and the cyclic evaluation index are integrated to obtain a comprehensive evaluation index, and the temperature rate of the PCR instrument in the next reaction stage is controlled by the comprehensive evaluation index.

[0118] In this embodiment, because the last sub-reaction stage is an extension stage, which involves the adjustment of the next cycle, the evaluation indicators of the three reaction stages in this cycle are integrated, and the temperature rate of the PCR instrument in the next reaction stage is controlled by the comprehensive evaluation indicator. Different comprehensive evaluation indicators correspond to different correction factors, which are controlled by the correction factor * the initial value of the temperature rate of the PCR instrument in the next reaction stage.

[0119] Through the description of the above implementation methods, those skilled in the art can clearly understand that the present invention can be implemented by hardware, or by software plus a necessary general hardware platform. Based on such an understanding, the technical solution of the present invention can be embodied in the form of a software product, which can be stored in a non-volatile storage medium (which can be a CD-ROM, a USB flash drive, a mobile hard disk, etc.), including a number of instructions for a computer device (which can be a personal computer, a server, or a network device, etc.) to execute the methods described in each implementation scenario of the present invention.

[0120] Compared with the prior art, the present invention has the following beneficial effects:

[0121] 1. Determine the reaction stage target information and the control cycle corresponding to each reaction stage based on the auxiliary reaction information and the information of the sample to be processed, so as to configure the reaction stage target information to ensure the accuracy of the reaction stage monitoring, and set the control cycle to carry out detailed control of the subsequent stages.

[0122] 2. Generate the deviation value and comprehensive influence coefficient of the reaction system for each sub-reaction stage based on real-time reaction data and real-time auxiliary reaction data. Use the deviation value and comprehensive influence coefficient of the reaction system to describe the fluctuation situation, and adjust the temperature rate of the PCR instrument in the next sub-reaction stage to improve the control effect and adaptability within a single stage.

[0123] 3. Control the temperature rate of the PCR instrument in the next reaction stage according to the reaction stage evaluation index and type of the previous sub-reaction stage. Describe the reaction effect of each reaction stage through the reaction stage evaluation index and type, and adjust the temperature rate of the PCR instrument in the next reaction stage to improve the control accuracy and adaptability between multiple stages.

[0124] Correspondingly, the present application also provides a PCR instrument self-optimizing control system, such as Figure 2 As shown, including,

[0125] A setting module is used to obtain the information of the sample to be processed and the basic information of the PCR instrument, configure the auxiliary reaction information according to the information of the sample to be processed and the basic information of the PCR instrument, and determine the reaction stage target information and the control cycle corresponding to each reaction stage according to the auxiliary reaction information and the information of the sample to be processed;

[0126] An analysis module is used to split each reaction stage into multiple sub-reaction stages according to the control cycle, configure the reaction stage target information on each sub-reaction stage, monitor the real-time reaction data and real-time auxiliary reaction data of each sub-reaction stage on the PCR instrument, and generate a deviation value and a comprehensive influence coefficient of the reaction system for each sub-reaction stage based on the real-time reaction data and the real-time auxiliary reaction data;

[0127] The first optimization module is used to control the temperature rate of the PCR instrument in the next sub-reaction stage by combining the deviation value of the previous sub-reaction stage and the comprehensive coefficient of the reaction system, so as to complete the optimization control of a single reaction stage;

[0128] The second optimization module is used to define the reaction stage evaluation index and the evaluation index type according to the overall reaction data of the reaction stage after each reaction stage is completed, and control the temperature rate of the PCR instrument of the next reaction stage according to the reaction stage evaluation index and type of the previous sub-reaction stage, so as to complete the continuous optimization control between multiple reaction stages;

[0129] Among them, the reaction stages involved in the PCR instrument include denaturation stage, annealing stage and extension stage.

[0130] Those skilled in the art will appreciate that the accompanying drawings are merely schematic diagrams of a preferred implementation scenario, and the modules or processes in the accompanying drawings are not necessarily required for implementing the present invention.

[0131] Those skilled in the art will appreciate that the modules in the device in the implementation scenario can be distributed in the device in the implementation scenario according to the implementation scenario description, or can be changed accordingly and located in one or more devices different from the implementation scenario. The modules in the above implementation scenario can be combined into one module, or can be further split into multiple sub-modules.

[0132] The above description is only a preferred specific implementation manner of the present invention, but the protection scope of the present invention is not limited thereto. Any technician familiar with the technical field can make equivalent replacements or changes according to the technical scheme and inventive concept of the present invention within the technical scope disclosed by the present invention, which should be covered by the protection scope of the present invention.

Claims

1. A PCR instrument self-optimization control method, characterized in that: include, Obtaining information of samples to be processed and basic information of the PCR instrument, configuring auxiliary reaction information according to the information of samples to be processed and the basic information of the PCR instrument, and determining reaction stage target information and a control cycle corresponding to each reaction stage according to the auxiliary reaction information and the information of samples to be processed; Each reaction stage is divided into multiple sub-reaction stages according to the control cycle, and the reaction stage target information is respectively configured on each sub-reaction stage, and the real-time reaction data and real-time auxiliary reaction data of each sub-reaction stage on the PCR instrument are monitored, and the deviation value and the comprehensive influence coefficient of the reaction system of each sub-reaction stage are generated based on the real-time reaction data and the real-time auxiliary reaction data; The temperature rate of the PCR instrument in the next sub-reaction stage is controlled by combining the deviation value of the previous sub-reaction stage and the comprehensive coefficient of the reaction system, so as to complete the optimization control of a single reaction stage; After each reaction stage is completed, the reaction stage evaluation index and evaluation index type are defined according to the overall reaction data of the reaction stage, and the temperature rate of the PCR instrument of the next reaction stage is controlled according to the reaction stage evaluation index and type of the previous sub-reaction stage, so as to complete the continuous optimization control between multiple reaction stages; Among them, the reaction stages involved in the PCR instrument include the denaturation stage, the annealing stage, and the extension stage; And determine the reaction stage target information and the control cycle corresponding to each reaction stage according to the auxiliary reaction information and the sample information to be processed, including: The target information of the reaction stage includes the target time and target temperature, and the auxiliary reaction information and the information of the sample to be processed include time-related information, temperature-related information and other information; Find similar reaction records in the historical reaction records through the information of the sample to be processed and the auxiliary reaction information, record them as similar reaction records, and determine the first target time range and the first target temperature range corresponding to the denaturation stage, the annealing stage and the extension stage respectively according to these similar reaction records; Comprehensively determine the second target time range and the second target temperature range corresponding to the denaturation stage, the annealing stage, and the extension stage, respectively, according to the time-related information and the temperature-related information in the auxiliary reaction information and the information of the sample to be processed; The intersection range of the first target duration range and the second target duration range is used as the target duration of each reaction stage, and the intersection range of the first target temperature range and the second target temperature range is used as the target temperature of each reaction stage; The control cycles corresponding to the denaturation phase, the annealing phase and the extension phase are set according to the auxiliary reaction information, the sample to be processed information and the reaction phase target information.

2. The PCR instrument self-optimization control method according to claim 1, characterized in that: According to the auxiliary reaction information, the sample information to be processed, and the target information of the reaction stage, the control cycles corresponding to the denaturation stage, the annealing stage, and the extension stage are set, including: In similar reaction records, each reaction stability index in the auxiliary reaction information and the sample to be processed information is evaluated, each reaction stability index is normalized, and a target parameter change curve of the target parameter of each reaction stage in the denaturation stage, the annealing stage, and the extension stage is constructed according to the similar reaction records, the target parameter change curve is split into a plurality of time-continuous partial curves, the average value of the slope change on each partial curve is calculated, and the average value of the slope change on each partial curve is combined to determine the average value of the slope change in the reaction stage; The stability level corresponding to the denaturation stage, annealing stage and extension stage is determined according to the target time, the average value of the slope change of the reaction stage and the reaction stability index; ； in, is the stability level, is the number of reaction stability indicators in the auxiliary reaction information and the sample information to be processed, The auxiliary reaction information and the sample information to be processed The weight of each reaction stability index, The auxiliary reaction information and the sample information to be processed The size of the reaction stability index, is the average value of the slope change during the reaction phase, For stable duration, It represents the stable duration mapped from the average value of the slope change in the reaction phase. is the median of the target duration, are the conversion coefficients of the stable duration and the target duration, respectively. are the first constant and the second constant respectively, [ ] is the rounding symbol; The control period corresponding to each reaction stage is set by the stability level, and different stability levels correspond to different control period sizes.

3. The PCR instrument self-optimization control method according to claim 1, characterized in that: Each reaction stage is divided into multiple sub-reaction stages according to the control cycle, and the reaction stage target information is configured on each sub-reaction stage, including: The corresponding reaction stage is divided into multiple sub-reaction stages according to the size of the control cycle, and the multiple sub-reaction stages are continuous in time. The sub-target temperature and sub-target fluorescence signal amount corresponding to each sub-reaction stage are set according to the target duration and target temperature of each reaction stage.

4. The PCR instrument self-optimization control method according to claim 3, characterized in that: Based on the real-time reaction data and real-time auxiliary reaction data, the deviation value of each sub-reaction stage and the comprehensive influence coefficient of the reaction system are generated, including: Compare the deviations of the real-time reaction data in each sub-reaction stage with the sub-target temperature and the sub-target fluorescence signal, generate a first deviation and a second deviation, and determine the deviation value of each sub-reaction stage by combining the first deviation and the second deviation; The real-time reaction data and the real-time auxiliary reaction data are input into the preset reaction system evaluation model to obtain the comprehensive influence coefficient of the reaction system.

5. The PCR instrument self-optimization control method according to claim 1, characterized in that: The temperature rate of the PCR instrument in the next sub-reaction stage is controlled by combining the deviation value of the previous sub-reaction stage and the comprehensive coefficient of the reaction system, including: Detect whether there is non-specific amplification in the previous sub-reaction stage. If there is non-specific amplification in the previous sub-reaction stage, identify the properties of the non-specific amplification, and control the temperature rate of the PCR instrument in the next sub-reaction stage according to the properties of the non-specific amplification, the deviation value and the comprehensive coefficient of the reaction system; ； in, For the The target value of the PCR instrument temperature rate for each sub-reaction stage, For the The initial value of the PCR instrument temperature rate for each sub-reaction stage, For the The comprehensive coefficient of the reaction system in each sub-reaction stage is It is The deviation value of each sub-reaction stage, are the conversion factor of the bias value and nonspecific amplification, is the number of attribute features of nonspecific amplification, For the The weight of the attribute features, For the The non-specific amplification of the sub-reaction stage The size of the attribute feature, is the third constant.

6. The PCR instrument self-optimization control method according to claim 1, characterized in that: After each reaction stage is completed, the reaction stage evaluation indicators and evaluation indicator types are defined according to the overall reaction data of the reaction stage, including: Determine the reaction stage evaluation index based on the temperature and fluorescence signal in the overall reaction data of the reaction stage; Evaluation indicator types include non-circular evaluation indicators and cyclic evaluation indicators; The reaction stage evaluation indexes of the denaturation stage and the annealing stage are defined as non-cyclic evaluation indexes, and the evaluation index of the extension stage is defined as cyclic evaluation index.

7. The PCR instrument self-optimization control method according to claim 6, characterized in that: And control the temperature rate of the PCR instrument in the next reaction stage according to the reaction stage evaluation index and type of the previous sub-reaction stage, including, When the previous sub-reaction stage is a denaturation stage or an annealing stage, the temperature rate of the PCR instrument in the next reaction stage is directly controlled according to the non-circulation evaluation index corresponding to each stage; When the previous sub-reaction stage is the extension stage, the non-cyclic evaluation index and the cyclic evaluation index are integrated to obtain a comprehensive evaluation index, and the temperature rate of the PCR instrument in the next reaction stage is controlled by the comprehensive evaluation index.

8. A PCR instrument self-optimizing control system, characterized in that: Used to implement the PCR instrument self-optimization control method according to any one of claims 1 to 7, the system comprises: A setting module is used to obtain the information of the sample to be processed and the basic information of the PCR instrument, configure the auxiliary reaction information according to the information of the sample to be processed and the basic information of the PCR instrument, and determine the reaction stage target information and the control cycle corresponding to each reaction stage according to the auxiliary reaction information and the information of the sample to be processed; An analysis module is used to split each reaction stage into multiple sub-reaction stages according to the control cycle, configure the reaction stage target information on each sub-reaction stage, monitor the real-time reaction data and real-time auxiliary reaction data of each sub-reaction stage on the PCR instrument, and generate a deviation value and a comprehensive influence coefficient of the reaction system for each sub-reaction stage based on the real-time reaction data and the real-time auxiliary reaction data; The first optimization module is used to control the temperature rate of the PCR instrument in the next sub-reaction stage by combining the deviation value of the previous sub-reaction stage and the comprehensive coefficient of the reaction system, so as to complete the optimization control of a single reaction stage; The second optimization module is used to define the reaction stage evaluation index and the evaluation index type according to the overall reaction data of the reaction stage after each reaction stage is completed, and control the temperature rate of the PCR instrument of the next reaction stage according to the reaction stage evaluation index and type of the previous sub-reaction stage, so as to complete the continuous optimization control between multiple reaction stages; Among them, the reaction stages involved in the PCR instrument include denaturation stage, annealing stage and extension stage.

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