CN114622006A - Nucleic acid temperature-changing amplification system based on 12V voltage drive - Google Patents
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Abstract
The invention relates to a nucleic acid temperature-changing amplification system based on 12V voltage driving, and belongs to the technical field of data processing. The system includes a memory and a processor that executes a computer program stored by the memory to implement the steps of: acquiring the fluorescence brightness value and temperature value of each EP tube in the PCR process of the nucleic acid temperature-variable amplification system to be judged, and constructing a fluorescence intensity sequence and a temperature sequence corresponding to each circle of an EP tube tray; calculating a local structural index corresponding to the nucleic acid temperature-variable amplification system according to the sequence; inputting the local structural index corresponding to the nucleic acid temperature-changing amplification system into a trained support vector machine classifier, and judging whether the nucleic acid temperature-changing amplification system has abnormal temperature control by using the trained support vector machine classifier. The invention realizes the real-time judgment of whether the temperature control of the nucleic acid temperature-variable amplification system is abnormal in the PCR process, and can be used for solving the problem of temperature abnormality in the PCR process.
Description
Technical Field
The invention relates to the technical field of data processing, in particular to a nucleic acid temperature-variable amplification system based on 12V voltage driving.
Background
Polymerase Chain Reaction (PCR) is an in vitro nucleic acid amplification technique developed in the middle of the 80 s. It has the outstanding advantages of specificity, sensitivity, high yield, rapidness, simplicity, good repeatability, easy automation and the like; the target gene or a certain DNA fragment to be researched can be amplified to hundreds of thousands to millions of times in a plurality of hours in a test tube, so that the target gene or the certain DNA fragment can be directly observed and judged by naked eyes; CN2011203905021U discloses a method for observing PCR process based on ultraviolet light, placing a reaction tube with fluorescence labeling amplification product on a reaction tube module of a heating/refrigerating device, connecting a PCR amplification instrument with a power supply, a host and a display, setting temperature and time after the power supply is turned on, carrying out denaturation, annealing and extension processes on DNA in the amplification reaction tube along with the rise and fall of the temperature, changing the color in the reaction tube under the action of ultraviolet light, and reacting on the display through a camera, so that the whole amplification process of the DNA can be intuitively known. Current methods of PCR process analysis using fluorescence are feasible and easy to implement.
At present, popular PCR equipment can heat and cool a reaction container through a plurality of Peltier elements, and a constant temperature and cold heat treatment process is controlled by a PID algorithm of an MCU, and the equipment can cause the reduction of temperature consistency along with the increase of service time and the problem of abnormal temperature due to temperature overshoot in the PID temperature regulation process.
Disclosure of Invention
In order to solve the problem of temperature anomaly in the existing PCR equipment in the PCR process, the invention aims to provide a nucleic acid temperature-variable amplification system based on 12V voltage driving, which comprises a memory and a processor, wherein the processor executes a computer program stored in the memory to realize the following steps:
acquiring the fluorescence brightness value and the temperature value of each EP tube in the PCR process of the nucleic acid temperature-variable amplification system to be judged, and constructing a fluorescence intensity sequence corresponding to each circle of the EP tube tray according to the position and the fluorescence brightness value of each EP tube in the EP tube tray; constructing a temperature sequence corresponding to each circle of the EP pipe tray according to the position and the temperature value of each EP pipe in the EP pipe tray; calculating a local structural index corresponding to the nucleic acid temperature-variable amplification system according to the fluorescence intensity sequence and the temperature sequence corresponding to each circle of the EP tube tray;
inputting the local structural index corresponding to the nucleic acid temperature-changing amplification system into a trained support vector machine classifier, and judging whether the nucleic acid temperature-changing amplification system has abnormal temperature control by using the trained support vector machine classifier;
the training process of the support vector machine classifier comprises the following steps: judging whether the local structural index of each training sample of the nucleic acid temperature-variable amplification system is abnormal or not based on a phase space analysis method, and marking each training sample of the nucleic acid temperature-variable amplification system; and training the support vector machine classifier based on the labeled training samples of the nucleic acid temperature-variable amplification systems.
Further, the determining whether the local structural index of each training sample of the nucleic acid temperature-variable amplification system is abnormal based on a phase-space analysis method includes:
for any nucleic acid temperature-variable amplification system training sample:
calculating the standard deviation of the tracking index corresponding to each observation time of the nucleic acid temperature-variable amplification system in the PCR process by taking the set time as a phase space;
calculating the structural separation index corresponding to each observation time according to the standard deviation of the tracking index corresponding to each observation time;
and judging whether the nucleic acid temperature-variable amplification system has the condition of structural separation index increase for more than two times in the PCR process, and if so, judging that the local structural index of the nucleic acid temperature-variable amplification system is abnormal.
Further, the structural separation index is 6 times the standard deviation of the tracking index.
Further, when the judgment results of the continuous set times are all abnormal temperature control, the abnormal temperature control of the nucleic acid temperature-variable amplification system is judged, and when the abnormal temperature control of the nucleic acid temperature-variable amplification system is judged, the multiplying power parameter in the PID temperature control system of the nucleic acid temperature-variable amplification system is controlled to be reduced until the judgment result of the nucleic acid temperature-variable amplification system is abnormal temperature control.
Further, the calculating the corresponding local structural index of the nucleic acid temperature-variable amplification system according to the fluorescence intensity sequence and the temperature sequence corresponding to each circle of the EP tube tray comprises:
calculating the local structural index of each circle by using the following formula, and calculating the corresponding local structural index of the nucleic acid temperature-variable amplification system according to the local structural index of each circle:
wherein,
is the local structural index of the ith turn,
is the temperature sequence corresponding to the ith turn,
the corresponding fluorescence intensity sequence of the ith circle is shown, STD is standard deviation, range is range, F is diagonal sampling function,
the number of symmetrical EP tube pairs corresponding to the ith turn is indicated.
Further, the classification result of the support vector machine classifier comprises normal temperature control quality and slightly abnormal temperature control quality.
Furthermore, each observation time includes a plurality of tracking indicators of the phase space at the acquisition time, and the standard deviation of the tracking indicator corresponding to each observation time is the standard deviation of the tracking indicator of the phase space at each acquisition time in each observation time.
Further, the observation time is 60s, and the interval between two adjacent acquisition moments is 10 s.
The invention has the beneficial effects that: the method comprises the steps of calculating a local structural index corresponding to the nucleic acid temperature-variable amplification system based on the fluorescence brightness value and the temperature value of each EP tube in the PCR process of the nucleic acid temperature-variable amplification system to be judged, taking the local structural index as the input of a support vector machine classifier, and judging whether the temperature control of the nucleic acid temperature-variable amplification system to be judged is abnormal or not by utilizing the support vector machine classifier; the invention realizes the real-time judgment of whether the temperature control of the nucleic acid temperature-variable amplification system is abnormal in the PCR process, and can timely regulate the PID temperature control system of the nucleic acid temperature-variable amplification system in the later period, thereby solving the problem of temperature abnormality in the PCR process.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions and advantages of 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 other drawings can be obtained by those skilled in the art without creative efforts.
FIG. 1 is a schematic diagram of an EP tube tray corresponding to a 12V voltage-driven nucleic acid temperature-variable amplification system according to the present invention;
FIG. 2 is a flowchart of the method for determining the quality of PCR temperature control based on a 12V voltage-driven nucleic acid temperature-variable amplification system according to the present invention.
Detailed Description
For further explanation of the present invention, the following detailed description is provided with reference to the drawings and preferred embodiments.
This example is directed to a 12V-based temperature-variable nucleic acid amplification system with a large number of EP tube trays, as shown in FIG. 1. Each slot of the EP tube tray is integrated with an ultraviolet light source and a photosensor for reading the fluorometrically measured PCR process. The bottom of each notch of the EP pipe tray is provided with a Peltier element and a temperature control sensor, the Peltier element can conduct the Peltier effect of heating and cooling to samples in each EP pipe through the EP pipe tray, and the temperature control sensor can read the temperature near the pipe wall of each EP pipe.
The nucleic acid temperature-variable amplification system driven by 12V voltage of the embodiment comprises a memory and a processor, wherein the processor executes a computer program stored in the memory to realize the PCR temperature control quality judgment method. As shown in fig. 2, the method for determining the PCR temperature control quality comprises the following steps:
step 1, acquiring a fluorescence brightness value and a temperature value of each EP tube in a PCR process of a nucleic acid temperature-variable amplification system to be judged, and constructing a fluorescence intensity sequence corresponding to each circle of an EP tube tray according to the position and the fluorescence brightness value of each EP tube in the EP tube tray; constructing a temperature sequence corresponding to each circle of the EP pipe tray according to the position and the temperature value of each EP pipe in the EP pipe tray; calculating a local structural index corresponding to the nucleic acid temperature-variable amplification system according to the fluorescence intensity sequence and the temperature sequence corresponding to each circle of the EP tube tray;
in order to analyze each sample of the EP tube tray, this example proposes a method of calculating a structural index of whether the PCR process is locally homogeneous or not by combining a fluorescence matrix and a temperature matrix. The specific process is as follows:
firstly, reading related sensor readings at the same time for each EP pipe notch;
reading the fluorescence brightness of the EP pipe notch after being irradiated by ultraviolet rays, wherein the unit is lux; the temperature values near the EP tube notch are read for estimating the tube wall temperature in degrees celsius.
Secondly, constructing a fluorescence intensity matrix based on the shape of the EP tube tray
And temperature matrix
Based on a fluorescence intensity matrix
And temperature matrix
Obtaining the corresponding fluorescence intensity sequence of each circle of the EP pipe tray
And temperature sequence
。
According to the shape of the EP pipe tray, a fluorescence intensity matrix can be constructed
And temperature matrix
The position of the reading for each EP tube in the matrix is the same as the actual position of the EP tube in the EP tube tray. Based on the matrix, the sensor reading of each EP tube is accessed in a shape like a Chinese character 'hui', and the mode of accessing in a shape like a Chinese character 'hui' is as follows: the sample from the center smallest unit, e.g., 2 ✕ 2, is expanded outward one turn, resulting in the outer turn of the sample (containing 4 x 4-2 x 2=12 samples), and so on, resulting in the sample per turn. When the width or height of one circle cannot be extended continuously, only the number of samples existing in the extended circle is calculated. The EP tube tray as shown in fig. 1, which includes six turns, with a first turn sample number of 4, a second turn sample number of 12, a third turn sample number of 6 x 6-4 x 4=20, and fourth, fifth and sixth turn sample numbers of 12 each.
Based on the method, the effect of each EP tube amplification caused by the heat conduction of the EP tube tray can be obtained. Since the EP tube tray should be a tray capable of uniformly conducting heat, a small portion of the peltier elements may not be well attached to the tray due to the high integration and frequent use of the EP tube tray, thereby causing non-uniformity of PCR process and temperature on the tray.
One-circle fluorescence intensity sequence can be obtained based on one circle of each visit
And temperature sequence
Where i is the index per turn.
Thirdly, for one circle of reading, calculating the local structural index w of each circleiThe calculation formula is as follows:
wherein F is a diagonal sampling function, and F acquires one element at a time and winds the elementThe subtended elements in the diagonal direction of the centrosymmetry, since the PCR environment should be uniform, the progress reflected by the fluorescence readings should also be as uniform as possible.
Represents the mean of the fluorescence readings of the phase difference of all two symmetrical EP tubes of the i-th circle,
the number of symmetrical EP tube pairs corresponding to the ith turn is indicated. When the average value is larger, the process is not uniform, so that the standard deviation of the temperature is increased, and the local structural index of the PCR process is reflected. When the index is too large, it means that one circle of the EP tube tray cannot be well amplified uniformly, and a phenomenon of local large difference is reflected.
So far, a local structural index vector w, w corresponding to the nucleic acid temperature-variable amplification system is obtained based on the local structural index of each circle
And N is the number of sampling circles of an EP tube tray of the current nucleic acid temperature-variable amplification system.
And 2, inputting the local structural index corresponding to the nucleic acid temperature-changing amplification system into a trained support vector machine classifier, and judging whether the nucleic acid temperature-changing amplification system has abnormal temperature control by using the trained support vector machine classifier.
The support vector machine classifier of the embodiment is used for judging whether the input local structural index is abnormal or not. In the embodiment, when the support vector machine classifier is trained, training samples need to be marked, and the marking types include normal and abnormal; the embodiment automatically analyzes whether the training samples are abnormal or not based on a phase space method, and if the analysis result is normal, the corresponding samples are marked as normal; and if the analysis result is abnormal, marking the corresponding sample as abnormal. Although the analysis of whether the PCR process is abnormal or not can be realized based on the phase space method, when the analysis of the abnormality is performed based on the phase space method, it is necessary to ensure continuous change of the phase space, and it is necessary to reverse the records corresponding to the next PCR process, and it is not possible to realize real-time judgment of whether the PCR process is abnormal or not. The following explains the relevant procedure.
Judging whether the local structural index of each training sample of the nucleic acid temperature-variable amplification system is abnormal or not based on a phase space analysis method, and marking each training sample of the nucleic acid temperature-variable amplification system;
for any training sample of the nucleic acid temperature-variable amplification system, the embodiment performs characterization on the local structural index of the nucleic acid temperature-variable amplification system, and then performs spatial domain expansion on the evolution of the temperature and the fluorescence in the amplification process according to a polar coordinate mode, thereby realizing an improved spatial warping (Wrapping) effect. In general phase space reconstruction, a uniform embedding mode is generally adopted, but the method cannot be applied to array data of PCR (polymerase chain reaction), so that edge effects and temperature overshoot phenomena can be analyzed based on a local structure, and the temperature unevenness phenomenon in the array can be continuously expressed by the systematic evolution rule of the temperature unevenness phenomenon.
The phase space is constructed such that the local structural index can be used as a space for all possible states of PCR quality. Under the observation of artificial participation, after ensuring that a PCR sample is normally loaded once, setting the starting time t to be 0, and establishing a vector of the PCR sample through local structurality
N is the number of local structures, a mutual information method is used for selecting a delay time parameter tau, a false adjacent point method is used for selecting an embedded dimension parameter m, and the phase space reconstruction mode is as follows:
...
...
at this point, the phase space of the N local structures with the local structural index change is reconstructed and used as the reference phase space
。
Each time the readings of the fluorescence intensity and temperature matrix are updated, a local structural index w for each substructure is obtained, and the data points w are updated
And N is the number of local structures, namely the number of sampling turns. The minimum analysis interval t is specified manually, 10 seconds in this embodiment. Using the space of reference phase
The same delay time τ and embedding dimension m reconstruct the phase space of the local structural index at time t, reconstructing the current phase space in the same way as described above:
...
...
so far, the phase space in the PCR process can be obtained
。
For a certain vector in the phase space at time t
Finding in reference phase space
The vector nearest to the vector
Wherein
。
Based on the above processing method, all data points w are updated every time sampling is performed, and the structure separation index within the production time interval T is obtained by taking T as the observation time interval
. Specifically, for the phase space, there are:
computing
And vector
The farthest distance is
Design phase space weights
. Let n increase by 1, continue to calculate
Up to
. Then, all tracking functions corresponding to all vectors in the phase space at the time t are utilized, and a tracking index of the phase space at the time t is calculated:
wherein q (n) is a weight function,
and m is the correlation dimension of the phase space at time t. Observing the phase space for a period of time, e.g. 60s, 6 times for P values, T =6, each P value corresponding to 10s, calculating
Average of T tracking indices
And standard deviation of
。
The tracking index reveals the amplification process per minimum unit
An indication of the phase space state of the course of the change over an observation time interval T. When the state index changes greatly in a period of time, the condition index means that the environment on which PCR depends in the production process changes obviously. Generally, when the PID typical error causes slow PCR progress difference change of each region, the local structural index of the amplification process fluctuates, and P does not change excessively. Therefore, standard deviation
Will be small so that the 3 sigma criterion can be used to estimate the tolerable quality change in PCR. This example, phase space analysis based on PCR, yields the structural separation index
。
This gave phase space analysis results in one PCR but was not applicable to multiple runs. Because the tracking index is in the ending period, each index of the system is different from that when a brand-new PCR sample is put, for example, the temperature of the brand-new PCR sample may be lower, while the temperature of the sample at the ending period may be higher, so that the change of the phase space is required to be ensured to be continuous, and the time reverse order processing is carried out on the next PCR data, so that the two PCRs form a cycle, the phase space analysis is ensured to be continuous and infinite, the structure separation index is analyzed more accurately, and the influence caused by reopening the PCR is avoided. Therefore, according to the above method, when performing PCR next time, all records need to be processed in reverse order, and the last sample is taken as the time when t is 0, so as to ensure the continuous change process, and so on, all analyses are performed alternately in positive and reverse order, thereby forming a cycle.
Although the phase space can track the PCR process to find the abnormality, since a cyclic process needs to be constructed, not every process can be performed in real time, and therefore the present embodiment performs SVM sample construction based on two classes of the structure discrete code and initializes the SVM, and the specific process is as follows:
and determining data influencing the PCR temperature control quality, and constructing a corresponding local structural index w.
The structure discrete code is a vector of the local structural index represented by the EP pipe pallet at each moment, and the structure separation index L corresponding to the EP pipe pallet can be calculated based on the vector of the local structural index represented by the EP pipe pallet at each moment.
And obtaining two levels of structure discrete codes based on L, and marking the structure discrete codes with high dispersion and the structure discrete codes with low dispersion. The marking process is as follows: index of structural separation during amplification
After two measurements and updates, the value is larger than the previous value L, which means that three adjacent structural discrete codes can reflect the phenomenon that PCR temperature control is abnormal, and the value is marked as B. Otherwise, the other samples are labeled as A, which indicates that the process is normal.
The sample comprises all encountered data conditions in PCR temperature control, is approximately complete data, the class of the sample is determined, an SVM classifier is trained based on the class, the quality effect of the PCR temperature control is divided into two classes by the classifier, wherein the PCR temperature control normal type A, PCR temperature control small abnormal type B exists.
Secondly, training the support vector machine classifier to obtain a trained support vector machine classifier;
the embodiment is based on two types of automatic analysis, adopts a supervised classification method, uses a Support Vector Machine (SVM) classifier, and classifies based on the characteristic parameters for expressing the PCR temperature control effect. The SVM classifier has the advantages that linear or nonlinear classification can be performed in a hyperplane and kernel function mode, and the result is accurate, so that the detection and classification method for the PCR temperature control effect abnormity based on the fuzzy multi-class SVM is used in the embodiment.
The calculation process of the SVM in this embodiment is: dividing hyperplanes with different abnormal degrees, calculating intervals among different abnormalities, analyzing hyperplane conditions when the intervals are maximum, and summarizing an optimal hyperplane. And classifying the compounded rubber data after the PCR temperature control is finished. The detailed operation steps belong to the prior art and are not described in detail.
Eighty percent of the obtained PCR temperature control quality data samples are used as training samples, the rest twenty percent of the obtained PCR temperature control quality data samples are used as test samples, a SVM classifier is carried out by using the PCR temperature control quality training sample data, the value of the classifier parameters is changed, various corresponding parameter values when the classification performance of the PCR temperature control effect classifier reaches the best are calculated, the classifier finishes parameter training, the rest twenty percent of the PCR temperature control data test samples are put into the classifier, the classification effect is tested, whether the classification is correct or not is judged, whether the correct rate meets the requirement or not is judged, otherwise, the parameters of the PCR temperature control quality classifier are continuously modified until the correct rate meets the use requirement.
Therefore, a trained SVM (support vector machine) classifier can be obtained, and two classifications of PCR temperature control quality can be realized based on the classifier, wherein one classification is a normal condition and the other classification is a slight abnormal condition. Since a serious abnormal situation is generally unlikely to occur, the present embodiment does not consider a serious abnormal situation.
And thirdly, inputting the local structural index corresponding to the nucleic acid temperature-changing amplification system to be judged into a trained support vector machine classifier, and judging whether the nucleic acid temperature-changing amplification system has abnormal temperature control by using the support vector machine classifier.
And (3) putting the local structural index corresponding to the nucleic acid temperature-changing amplification system to be judged into an SVM (support vector machine) for 2 classification by calculation to obtain the result of whether the temperature-changing amplification system has abnormal temperature control. The embodiment reduces the time cost of manual observation, and solves the problem of how to reasonably judge the temperature control abnormity of a plurality of PCR samples in all possibilities of a chaotic system in the space of temperature and PCR processes.
And based on the abnormal classification result, carrying out dynamic adjustment on the PCR temperature control parameters and calculating the stability of the PCR temperature control effect. In particular, the situation is marked as normal
Minor abnormal marksIs composed of
To a
For similar PCR samples, the temperature control parameters for the subsequent PCR temperature control process do not need to be adjusted. And for distributing to
In the PCR temperature control process of the group, slight PCR progress abnormity occurs in the PCR temperature control process, possibly the temperature of the PCR temperature control is abnormal due to the heat preservation and temperature change performance, for example, the temperature is uneven due to the aging of the joint of the original Peltier element and the heat exchanger, the embodiment improves the subsequent PCR temperature control quality by dynamic scaling of the multiplying power K parameter of the PID, and the reason for changing the multiplying power K is that the overshoot phenomenon may occur when the multiplying power K is too large; when the B group condition appears continuously for a plurality of times in a PCR process, the multiplying power K is controlled to be attenuated to another new lower value preset by an implementer until the SVM allocates a new structure discrete code to the A group.
In this embodiment, a local structural index corresponding to the nucleic acid temperature-variable amplification system is calculated based on a fluorescence brightness value and a temperature value of each EP tube of the nucleic acid temperature-variable amplification system to be determined in a PCR process, the local structural index is used as an input of a support vector machine classifier, and the support vector machine classifier is used to determine whether the nucleic acid temperature-variable amplification system to be determined is abnormal in temperature control; the invention realizes the real-time judgment of whether the temperature control of the nucleic acid temperature-variable amplification system is abnormal in the PCR process, and can timely regulate the PID temperature control system of the nucleic acid temperature-variable amplification system in the later period, thereby solving the problem of temperature abnormality in the PCR process.
The above-mentioned embodiments are only used for illustrating the technical solutions of the present application, and not for limiting the same; although the present application has been described in detail with reference to the foregoing embodiments, it should be understood by those of ordinary skill in the art that: the technical solutions described in the foregoing embodiments may still be modified, or some technical features may be equivalently replaced; such modifications and substitutions do not substantially depart from the spirit and scope of the embodiments of the present application and are intended to be included within the scope of the present application.
Claims (8)
1. A 12V voltage-driven nucleic acid temperature-variable amplification system, comprising a memory and a processor, wherein the processor executes a computer program stored in the memory to implement the following steps:
acquiring the fluorescence brightness value and the temperature value of each EP tube in the PCR process of the nucleic acid temperature-variable amplification system to be judged, and constructing a fluorescence intensity sequence corresponding to each circle of the EP tube tray according to the position and the fluorescence brightness value of each EP tube in the EP tube tray; constructing a temperature sequence corresponding to each circle of the EP pipe tray according to the position and the temperature value of each EP pipe in the EP pipe tray; calculating a local structural index corresponding to the nucleic acid temperature-variable amplification system according to the fluorescence intensity sequence and the temperature sequence corresponding to each circle of the EP tube tray;
inputting the local structural index corresponding to the nucleic acid temperature-changing amplification system into a trained support vector machine classifier, and judging whether the nucleic acid temperature-changing amplification system has abnormal temperature control by using the trained support vector machine classifier;
the training process of the support vector machine classifier comprises the following steps: judging whether the local structural index of each training sample of the nucleic acid temperature-variable amplification system is abnormal or not based on a phase space analysis method, and marking each training sample of the nucleic acid temperature-variable amplification system; and training the support vector machine classifier based on the labeled training samples of the nucleic acid temperature-variable amplification systems.
2. The system according to claim 1, wherein the determining whether the local structural index of the training sample of each nucleic acid temperature-variable amplification system is abnormal based on the phase-space analysis method comprises:
for any nucleic acid temperature-variable amplification system training sample:
calculating the standard deviation of the tracking index corresponding to each observation time of the nucleic acid temperature-variable amplification system in the PCR process by taking the set time as a phase space;
calculating the structural separation index corresponding to each observation time according to the standard deviation of the tracking index corresponding to each observation time;
and judging whether the nucleic acid temperature-changing amplification system has the condition of structural separation index increase for more than two times in the PCR process, and if so, judging that the local structural index of the nucleic acid temperature-changing amplification system is abnormal.
3. The nucleic acid temperature-variable amplification system based on 12V voltage drive of claim 2, wherein the structure separation index is a standard deviation of a 6-fold tracking index.
4. The 12V voltage-driven nucleic acid temperature-variable amplification system according to claim 1, wherein the temperature-variable amplification system is determined to have abnormal temperature control when all the results of determination for the number of consecutive times are abnormal temperature control, and wherein the PID temperature control system of the nucleic acid temperature-variable amplification system is controlled to decrease the magnification parameter until the result of determination for the nucleic acid temperature-variable amplification system is abnormal temperature control when it is determined that the temperature-variable amplification system has abnormal temperature control.
5. The system for nucleic acid temperature-variable amplification based on 12V voltage driving according to claim 1, wherein the calculating of the corresponding local structural index of the nucleic acid temperature-variable amplification system according to the fluorescence intensity sequence and the temperature sequence corresponding to each circle of the EP tube tray comprises:
calculating the local structural index of each circle by using the following formula, and calculating the corresponding local structural index of the nucleic acid temperature-changing amplification system according to the local structural index of each circle:
wherein,
is the local structural index of the ith turn,
is a temperature sequence corresponding to the ith turn,
the corresponding fluorescence intensity sequence of the ith circle is shown, STD is standard deviation, range is range, F is diagonal sampling function,
the number of symmetrical EP tube pairs corresponding to the ith turn is indicated.
6. The 12V voltage-driven nucleic acid temperature-variable amplification system according to claim 1, wherein the classification result of the support vector machine classifier comprises both temperature-controlled quality normality and temperature-controlled quality slight abnormality.
7. The system according to claim 2, wherein each observation period comprises a plurality of phase space tracking indicators at the collection time, and the standard deviation of the tracking indicator corresponding to each observation period is the standard deviation of the phase space tracking indicator at each collection time in each observation period.
8. The system according to claim 7, wherein the observation time is 60s, and the interval between two adjacent collection times is 10 s.
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