Method for determining cycle threshold, electronic device, and medium
By performing linear regression processing and residual standard error analysis on the amplification curves of fluorescent polymerase chain reaction (PCR), the baseline and threshold line were determined, which solved the problem of low accuracy of the cycling threshold in the non-amplification state of PCR reagents and improved the accuracy of the cycling threshold.
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Patents(China)
- Current Assignee / Owner
- SHENZHEN EVERBEST MACHINERY IND
- Filing Date
- 2023-02-01
- Publication Date
- 2026-07-07
Smart Images
Figure CN116153419B_ABST
Abstract
Description
Technical Field
[0001] This application belongs to the field of biological detection technology, and in particular relates to a method for determining a cycle threshold, an electronic device, and a medium. Background Technology
[0002] Real-time polymerase chain reaction (Real-time PCR) is a biological technique that involves adding a fluorescent group to a polymerase chain reaction (PCR) system and using the fluorescence signal to monitor the entire PCR process. In practical applications, the fluorescence signal of each cycle product in the PCR reaction can be detected in real time to obtain the fluorescence signal amplification curve. Then, the baseline and threshold line corresponding to the amplification curve can be obtained. Finally, the cycle threshold (CT) can be obtained based on the threshold line and the amplification curve.
[0003] The methods used to obtain the baseline and threshold lines have a significant impact on CT scans. Currently, the commonly used method for obtaining the baseline is to use a straight line with a slope of 0 and an intercept equal to the average of the 3rd to 15th cycle fluorescence values as the baseline. Similarly, the commonly used method for obtaining the threshold line is to use a straight line with a slope of 0 and an intercept equal to the sum of the baseline intercept and the standard deviations of the 3rd to 15th cycle fluorescence values as the threshold line. While these baseline and threshold line acquisition methods are generally statistically significant, they may lose their statistical significance in certain special cases (e.g., when the fluorescent polymerase chain reaction reagent is in a non-amplified state, i.e., when the fluorescence signal shows a regular increase or decrease), thus reducing the accuracy of the obtained CT scan. Summary of the Invention
[0004] In view of this, embodiments of this application provide a method, electronic device, and medium for determining a cycling threshold. This addresses the technical problem of low accuracy in determining the cycling threshold using existing methods when the fluorescent polymerase chain reaction reagent is in a non-amplification state.
[0005] In a first aspect, embodiments of this application provide a method for determining a loop threshold, comprising:
[0006] Linear regression is performed on the fluorescence values from the Nth to the Mth cycles of the amplification curve to be processed to obtain the baseline corresponding to the amplification curve; where N and M are both positive integers, and M is greater than N;
[0007] Determine the residual standard error of the fluorescence values from the Nth cycle to the Mth cycle;
[0008] The threshold line corresponding to the amplification curve is determined based on the baseline and the residual standard error.
[0009] The cyclic threshold is determined based on the threshold line and the amplification curve.
[0010] Optionally, N is 3 and M is 15.
[0011] Optionally, determining the threshold line based on the baseline and the residual standard error includes:
[0012] Obtain the first slope and first intercept of the baseline;
[0013] The second slope of the threshold line is determined based on the first slope;
[0014] The second intercept of the threshold line is determined based on the first intercept and the residual standard error.
[0015] The threshold line is determined based on the second slope and the second intercept.
[0016] Optionally, determining the second slope of the threshold line based on the first slope includes:
[0017] The first slope is determined as the second slope of the threshold line.
[0018] Optionally, determining the second intercept of the threshold line based on the first intercept and the residual standard error includes:
[0019] The sum of the first intercept and the residual standard error of a preset multiple is determined as the second intercept of the threshold line.
[0020] Optionally, the preset multiple is greater than or equal to 10 and less than or equal to 30.
[0021] Optionally, the amplification curve consists of multiple cycle points, each cycle point being configured with a first cycle number corresponding to the cycle point and the fluorescence value, characterized in that determining the cycle threshold based on the threshold line and the amplification curve includes:
[0022] Determine multiple equally spaced second cycle numbers between each pair of adjacent first cycle numbers;
[0023] Determine the first comparison value corresponding to each of the second cycle numbers from the amplification curves;
[0024] Determine the second comparison value for each of the second cycle numbers from the threshold lines;
[0025] For any given second cycle number, if the absolute value of the difference between the first comparison value and the second comparison value corresponding to the second cycle number is less than or equal to a preset threshold, then the second cycle number is determined as the cycle threshold.
[0026] Secondly, embodiments of this application provide an electronic device, including:
[0027] The first determining unit is used to perform linear regression processing on the fluorescence values of the Nth to Mth cycles in the amplification curve to be processed, so as to obtain the baseline corresponding to the amplification curve; where N and M are both positive integers, and M is greater than N;
[0028] The second determining unit is used to determine the residual standard error of the fluorescence values from the Nth cycle to the Mth cycle in the amplification curve;
[0029] The third determining unit is used to determine the threshold line corresponding to the amplification curve based on the baseline and the residual standard error.
[0030] The fourth determining unit is used to determine the cycle threshold based on the threshold line and the amplification curve.
[0031] Thirdly, embodiments of this application provide an electronic device, including a memory, a processor, and a computer program stored in the memory and executable on the processor, characterized in that the processor, when executing the computer program, implements each step of the method for determining the loop threshold as described in any of the first aspects above.
[0032] Fourthly, embodiments of this application provide a computer-readable storage medium storing a computer program that, when executed by a processor, implements the steps of the method for determining the loop threshold as described in any of the first aspects above.
[0033] Fifthly, embodiments of this application provide a computer program product that, when run on a terminal device, causes the terminal device to perform the steps of the method described in the first aspect or any optional method of the first aspect.
[0034] The method for determining the cycle threshold, the electronic device, the computer-readable storage medium, and the computer program product provided in this application have the following beneficial effects:
[0035] The method for determining the cycle threshold provided in this application involves performing linear regression on the fluorescence values from the Nth to the Mth cycles of the amplification curve to obtain the baseline corresponding to the amplification curve; determining the residual standard error of the fluorescence values from the Nth to the Mth cycles; determining the threshold line corresponding to the amplification curve based on the baseline and the residual standard error; and determining the cycle threshold based on the threshold line and the amplification curve. Since the method for determining the baseline and threshold line corresponding to the amplification curve in this scheme remains statistically significant even when the fluorescent polymerase chain reaction reagent is in a non-amplification state, the method for determining the cycle threshold provided in this scheme also remains statistically significant even when the fluorescent polymerase chain reaction reagent is in a non-amplification state, thereby improving the accuracy of the obtained CT. Attached Figure Description
[0036] To more clearly illustrate the technical solutions in the embodiments of this application, the drawings used in the description of the embodiments or the prior art will be briefly introduced below. Obviously, the drawings described below are only some embodiments of this application. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.
[0037] Figure 1 A flowchart illustrating the implementation of a method for determining a loop threshold provided in an embodiment of this application;
[0038] Figure 2 A schematic diagram of the amplification curves provided in the embodiments of this application;
[0039] Figure 3 A schematic diagram of the baseline corresponding to an amplification curve provided in an embodiment of this application;
[0040] Figure 4 A flowchart illustrating the implementation of determining a threshold line based on baseline and residual standard error is provided in this application embodiment.
[0041] Figure 5 This is a schematic diagram of a threshold line corresponding to an amplification curve provided in an embodiment of this application;
[0042] Figure 6 A flowchart illustrating the implementation of determining the cyclic threshold based on a threshold line and an amplification curve, provided in this application embodiment;
[0043] Figure 7 This is a schematic diagram of the structure of an electronic device provided in an embodiment of this application;
[0044] Figure 8 This is a schematic diagram of the structure of an electronic device provided in an embodiment of this application. Detailed Implementation
[0045] It should be noted that the terminology used in the embodiments of this application is only for explaining specific embodiments of this application and is not intended to limit this application. In the description of the embodiments of this application, unless otherwise stated, "multiple" means two or more, "at least one" or "one or more" means one, two or more. The terms "first" and "second" are used for descriptive purposes only and should not be construed as indicating or implying relative importance or implicitly specifying the number of indicated technical features. Thus, a feature defined as "first" or "second" may explicitly or implicitly include one or more of that feature.
[0046] References to "one embodiment" or "some embodiments" as described in this specification mean that one or more embodiments of this application include a specific feature, structure, or characteristic described in connection with that embodiment. Therefore, the phrases "in one embodiment," "in some embodiments," "in other embodiments," "in still other embodiments," etc., appearing in different parts of this specification do not necessarily refer to the same embodiment, but rather mean "one or more, but not all, embodiments," unless otherwise specifically emphasized. The terms "comprising," "including," "having," and variations thereof mean "including but not limited to," unless otherwise specifically emphasized.
[0047] The execution subject of the method for determining the loop threshold provided in this application embodiment can be an electronic device. The electronic device may include, for example, a loop threshold determining device.
[0048] In this embodiment of the application, when a user wants to determine the CT corresponding to the amplification curve based on the amplification curve to be processed, the user can input the amplification curve into the electronic device. After obtaining the amplification curve, the electronic device can execute each step of the cycle threshold determination method provided in this embodiment of the application, so that it can still accurately determine the CT corresponding to the amplification curve when the fluorescent polymerase chain reaction reagent is in a non-amplification state, thereby improving the accuracy of CT determination.
[0049] Please see Figure 1 , Figure 1 The flowchart illustrates a method for determining a loop threshold provided in this application embodiment. This method may include steps S101 to S104, as detailed below:
[0050] In S101, the fluorescence values from the Nth to the Mth cycles of the amplification curve to be processed are linearly regressed to obtain the baseline corresponding to the amplification curve.
[0051] In this embodiment, the amplification curve can be used to describe the relationship between the first cycle number and the fluorescence value, where the first cycle number is the cycle number of the polymerase chain reaction. The amplification curve can consist of multiple cycle points. Each cycle point is configured with a corresponding first cycle number and fluorescence value; the first cycle number can be used to represent the number of cycles in the fluorescent polymerase chain reaction, and the first cycle number can be a positive integer; the fluorescence value can be used to represent the magnitude of the fluorescence signal in the fluorescent polymerase chain reaction, and the rules for determining the fluorescence value can be set based on actual conditions, and are not limited here. For example, if the first cycle number of cycle point A is B and the fluorescence value is C, it means that the fluorescence value of the Bth polymerase chain reaction cycle in the fluorescent polymerase chain reaction is C.
[0052] In practical applications, amplification curves can be obtained by collecting fluorescence signals once per cycle of the fluorescent polymerase chain reaction and plotting them on a coordinate system. The horizontal axis of this coordinate system can be the first cycle number, and the vertical axis can be the fluorescence value. For an example, please refer to [link to example]. Figure 2 , Figure 2 A schematic diagram of the amplification curves provided in the embodiments of this application, as shown below. Figure 2 As shown, Figure 2 Curve a in the figure is the amplification curve provided in the embodiments of this application, which consists of multiple cycle points.
[0053] In this embodiment, N and M are both positive integers, and M is greater than N. For example, N can be 3 and M can be 15. It should be noted that the specific values of N and M can also be set according to the needs of actual applications, and are not limited here.
[0054] The baseline is a straight line determined based on one or more polymerase chain reaction cycles in which the initial fluorescence value does not change significantly.
[0055] In this embodiment, when the electronic device needs to determine the CT corresponding to the amplification curve based on the amplification curve to be processed, the electronic device can first perform linear regression processing on the fluorescence values from the Nth polymerase chain reaction cycle to the Mth polymerase chain reaction cycle in the amplification curve to be processed, and obtain the baseline corresponding to the amplification curve. For example, please refer to... Figure 3 , Figure 3 This is a schematic diagram of the baseline corresponding to an amplification curve provided in an embodiment of this application. For example... Figure 3 As shown, Figure 3 The straight line b in the figure is the baseline provided in the embodiments of this application. This baseline is obtained by linear regression processing based on the fluorescence values of the Nth to Mth polymerase chain reaction cycles in the amplification curve.
[0056] In S102, the residual standard error of the fluorescence values from the Nth cycle to the Mth cycle is determined.
[0057] In the embodiments of this application, the residual standard error (RSE) is used to describe the average offset between each cycle point and the baseline between the Nth and Mth cycles.
[0058] In one possible implementation, after determining the baseline corresponding to the amplification curve, the electronic device can determine the residual standard error of the fluorescence values from the Nth to the Mth cycles based on the fluorescence values at each cycle point between the Nth and Mth cycles, the baseline, and the first equation. The first equation describes the correspondence between the fluorescence values, the baseline, and the residual standard error. This first equation can be determined in advance through multiple experimental measurements; no specific limitations are imposed on it here.
[0059] In S103, the threshold line corresponding to the amplification curve is determined based on the baseline and residual standard error.
[0060] In the embodiments of this application, the threshold line refers to a straight line determined based on the baseline and residual standard error, which can be used together with the amplification curve to determine the cyclic threshold.
[0061] After determining the baseline and residual standard error of the fluorescence value corresponding to the amplification curve, the electronic device can determine the threshold line corresponding to the amplification curve based on the baseline and the residual standard error.
[0062] In one possible implementation, S103 can be achieved by, for example... Figure 4 The implementations of S1031 to S1034 shown are described in detail below:
[0063] In S1031, the first slope and first intercept of the baseline are obtained.
[0064] In this implementation, when the electronic device needs to determine the threshold line based on the baseline and the residual standard error, it can first obtain the first slope and the first intercept of the baseline.
[0065] In S1032, the second slope of the threshold line is determined based on the first slope.
[0066] In this implementation, after obtaining the first slope of the baseline, the electronic device can determine the first slope of the baseline as the second slope of the threshold line.
[0067] In S1033, the second intercept of the threshold line is determined based on the first intercept and the residual standard error.
[0068] In this implementation, after obtaining the first intercept of the baseline, the electronic device can determine the second intercept of the threshold line by summing the first intercept with the residual standard error of a preset multiple. For example, the preset multiple can be greater than or equal to 10 and less than or equal to 30.
[0069] In S1034, the threshold line is determined based on the second slope and the second intercept.
[0070] In this implementation, after the electronic device determines the second slope and the second intercept of the threshold line, it can determine the threshold line based on the second slope and the second intercept. Specifically, the second slope can be used as the slope of the threshold line, and the second intercept can be used as the intercept of the threshold line, thereby obtaining the corresponding threshold line.
[0071] For example, please refer to Figure 5 , Figure 5 This is a schematic diagram of the threshold line provided in an embodiment of this application. Figure 5 The straight line c in the figure is a schematic diagram of a threshold line corresponding to an amplification curve provided in an embodiment of this application. This threshold line is obtained based on the baseline b and the residual standard error.
[0072] In S104, the cycle threshold is determined based on the threshold line and the amplification curve.
[0073] In this embodiment, the cycle threshold refers to the number of cycles required for the fluorescence value to reach a preset threshold in a fluorescent polymerase chain reaction. In practical applications, the cycle threshold can be determined based on the cycle number corresponding to the intersection of the amplification curve and the threshold line.
[0074] Once the threshold line corresponding to the amplification curve is determined, the electronic device can determine the cycle threshold based on the threshold line and the amplification curve.
[0075] In one possible implementation, S104 can be achieved by, for example... Figure 6 The implementations of S1041 to S1044 shown are described in detail below:
[0076] In S1041, multiple equally spaced second cycle numbers are determined from each pair of adjacent first cycle numbers.
[0077] In this implementation, the number of second cycle numbers between any two adjacent first cycle numbers can be set according to actual needs and is not limited here. For example, the number of second cycle numbers between any two adjacent first cycle numbers can be 100. Based on this, the second cycle numbers between first cycle number 0 and first cycle number 1 can be 0.00, 0.01, 0.02, ..., 0.99 in ascending order; the second cycle numbers between first cycle number 1 and first cycle number 2 can be 1.00, 1.01, 1.02, ..., 1.99 in ascending order; and so on.
[0078] In S1042, the first comparison value corresponding to each second cycle number is determined from the amplification curve.
[0079] In this implementation, the electronic device can substitute each second cycle number into the amplification curve equation to determine the first comparison value corresponding to each second cycle number. Specifically, the second cycle number can be substituted as the independent variable of the amplification curve equation, and the value of the dependent variable of the resulting amplification curve equation can be determined as the first comparison value corresponding to that second cycle number.
[0080] In S1043, the second comparison value for each second cycle number is determined from the threshold line.
[0081] In this implementation, the electronic device can substitute each second cycle number into the threshold line equation to determine the second comparison value corresponding to each second cycle number. Specifically, the second cycle number can be substituted into the threshold line equation as the independent variable, and the value of the dependent variable obtained from the threshold line equation is determined as the second comparison value corresponding to that second cycle number.
[0082] In S1044, for any second cycle number, if the absolute value of the difference between the first comparison value and the second comparison value corresponding to the second cycle number is less than or equal to a preset threshold, then the second cycle number is determined as the cycle threshold.
[0083] In this implementation, after determining the first comparison value and the second comparison value corresponding to each second cycle number, the electronic device can compare the absolute value of the difference between the first comparison value and the second comparison value corresponding to each second cycle number with a preset threshold. If the absolute value of the difference between the first comparison value and the second comparison value corresponding to a certain second cycle number is less than or equal to the preset threshold, then the second cycle number is determined as a cycle threshold. If the absolute value of the difference between the first comparison value and the second comparison value corresponding to a certain second cycle number is greater than the preset threshold, then the absolute value of the difference between the first comparison value and the second comparison value corresponding to the next second cycle number is compared with the preset threshold to determine whether the next second cycle number is a cycle threshold. This process continues until all second cycle numbers are determined to be cycle thresholds.
[0084] In practical applications, the preset threshold can be set based on actual needs, and no specific restrictions are made here.
[0085] As can be seen from the above, the method for determining the cycle threshold provided in this application involves performing linear regression on the fluorescence values from the Nth to the Mth cycles of the amplification curve to obtain the baseline corresponding to the amplification curve; determining the residual standard error of the fluorescence values from the Nth to the Mth cycles; determining the threshold line corresponding to the amplification curve based on the baseline and the residual standard error; and determining the cycle threshold based on the threshold line and the amplification curve. Since the method for determining the baseline and threshold line corresponding to the amplification curve in this scheme remains statistically significant even when the fluorescent polymerase chain reaction reagent is in a non-amplification state, the method for determining the cycle threshold provided in this scheme also remains statistically significant even when the fluorescent polymerase chain reaction reagent is in a non-amplification state, thereby improving the accuracy of the obtained CT.
[0086] Based on the method for determining the loop threshold provided in the above embodiments, this application further provides an electronic device for implementing the above method embodiments. Please refer to [link to relevant documentation]. Figure 7 , Figure 7 This is a schematic diagram of the structure of an electronic device provided in an embodiment of this application. Figure 7 As shown, the electronic device 70 may include a first determining unit 71, a second determining unit 72, a third determining unit 73, and a fourth determining unit 74. Wherein:
[0087] The first determining unit 71 is used to perform linear regression processing on the fluorescence values of the Nth to Mth cycles in the amplification curve to be processed, to obtain the baseline corresponding to the amplification curve. N and M are both positive integers, and M is greater than N.
[0088] The second determining unit 72 is used to determine the residual standard error of the fluorescence values from the Nth cycle to the Mth cycle in the amplification curve.
[0089] The third determining unit 73 is used to determine the threshold line corresponding to the amplification curve based on the baseline and the residual standard error.
[0090] The fourth determining unit 74 is used to determine the cycle threshold based on the threshold line and the amplification curve.
[0091] Optionally, N is 3 and M is 15.
[0092] Optionally, the third determining unit 73 may include a first acquiring unit, a fifth determining unit, a sixth determining unit, and a seventh determining unit. Wherein:
[0093] The first acquisition unit is used to acquire the first slope and the first intercept of the baseline.
[0094] The fifth determining unit is used to determine the second slope of the threshold line based on the first slope.
[0095] The sixth determining unit is used to determine the second intercept of the threshold line based on the first intercept and the residual standard error.
[0096] The seventh determining unit is used to determine the threshold line based on the second slope and the second intercept.
[0097] Optionally, the fifth determining unit is specifically used to determine the first slope as the second slope of the threshold line.
[0098] Optionally, the sixth determining unit is specifically used to determine the second intercept of the threshold line by summing the first intercept with the residual standard error of a preset multiple.
[0099] Optionally, the preset multiple is greater than or equal to 10 and less than or equal to 30.
[0100] Optionally, the fourth determining unit 74 may include an eighth determining unit, a ninth determining unit, a tenth determining unit, and an eleventh determining unit. Wherein:
[0101] The eighth determining unit is used to determine a plurality of equally spaced second cycle numbers between each pair of adjacent first cycle numbers.
[0102] The ninth determining unit is used to determine the first comparison value corresponding to each of the second cycle numbers from the amplification curve.
[0103] The tenth determining unit is used to determine the second comparison value of each of the second cycle numbers from the threshold lines.
[0104] The eleventh determining unit is used to determine the second cycle number as the cycle threshold if, for any second cycle number, the absolute value of the difference between the first comparison value and the second comparison value corresponding to the second cycle number is less than or equal to a preset threshold.
[0105] It should be noted that the information interaction and execution process between the above-mentioned units are based on the same concept as the method embodiments of this application. Their specific functions and technical effects can be referred to the method embodiments section, and will not be repeated here.
[0106] Please see Figure 8 , Figure 8 This is a schematic diagram of the structure of an electronic device provided in an embodiment of this application. Figure 8 As shown, the electronic device 8 provided in this embodiment may include: a processor 80, a memory 81, and a computer program 82 stored in the memory 81 and executable on the processor 80. For example, a program corresponding to a method for determining a loop threshold. When the processor 80 executes the computer program 82, it implements the steps described above in the embodiment of the method for determining a loop threshold, for example... Figure 1 S101 to S104 shown Figure 4 S1031 to S1034 and shown Figure 6 S1041 to S1044 are shown. Alternatively, when the processor 80 executes the computer program 82, it implements the functions of each module / unit in the embodiment corresponding to the above-described electronic device 70, for example... Figure 7 The functions of units 71 to 74 shown.
[0107] For example, computer program 82 can be divided into one or more modules / units, one or more of which are stored in memory 81 and executed by processor 80 to complete this application. One or more modules / units can be a series of computer program instruction segments capable of performing specific functions, which describe the execution process of computer program 82 in electronic device 8. For example, computer program 82 can be divided into a first determining unit, a second determining unit, a third determining unit, and a fourth determining unit; the specific functions of each unit are described in [reference needed]. Figure 7 The relevant descriptions in the corresponding embodiments are not repeated here.
[0108] Those skilled in the art will understand that Figure 8 This is merely an example of electronic device 8 and does not constitute a limitation on electronic device 8. It may include more or fewer components than shown, or combine certain components, or use different components.
[0109] The processor 80 can be a central processing unit (CPU), or other general-purpose processors, digital signal processors (DSPs), application-specific integrated circuits (ASICs), field-programmable gate arrays (FPGAs), or other programmable logic devices, discrete gate or transistor logic devices, discrete hardware components, etc. The general-purpose processor can be a microprocessor or any conventional processor.
[0110] The memory 81 can be an internal storage unit of the electronic device 8, such as a hard disk or RAM. The memory 81 can also be an external storage device of the electronic device 8, such as a plug-in hard disk, smart media card (SMC), secure digital (SD) card, or flash card. Furthermore, the memory 81 can include both internal and external storage units of the electronic device 8. The memory 81 is used to store computer programs and other programs and data required by the electronic device. The memory 81 can also be used to temporarily store data that has been output or will be output.
[0111] Those skilled in the art will clearly understand that, for the sake of convenience and brevity, the above-described division of functional units is merely an example. In practical applications, the above functions can be assigned to different functional units as needed, that is, the internal structure of the electronic device 70 can be divided into different functional units to complete all or part of the functions described above. The functional units in the embodiments can be integrated into one processing unit, or each unit can exist physically separately, or two or more units can be integrated into one unit. The integrated unit can be implemented in hardware or as a software functional unit. Furthermore, the specific names of the functional units are only for easy differentiation and are not intended to limit the scope of protection of this application. The specific working process of the units in the above system can be referred to the corresponding process in the foregoing method embodiments, and will not be repeated here.
[0112] This application also provides a computer-readable storage medium storing a computer program that, when executed by a processor, can implement the steps in the various method embodiments described above.
[0113] This application provides a computer program product that, when run on an electronic device, enables the electronic device to perform the steps described in the various method embodiments above.
[0114] In the above embodiments, the descriptions of each embodiment have different focuses. For parts that are not described in detail or recorded in a certain embodiment, refer to the relevant descriptions of other embodiments.
[0115] In the above embodiments, the descriptions of each embodiment have different focuses. For parts that are not described in detail or recorded in a certain embodiment, refer to the relevant descriptions of other embodiments.
[0116] Those skilled in the art will recognize that the units and algorithm steps of the various examples described in conjunction with the embodiments disclosed herein can be implemented in electronic hardware, or a combination of computer software and electronic hardware. Whether these functions are implemented in hardware or software depends on the specific application and design constraints of the technical solution. Those skilled in the art can use different methods to implement the described functions for each specific application, but such implementation should not be considered beyond the scope of this application.
[0117] The above-described embodiments are only used to illustrate the technical solutions of this application, and are not intended to limit them. Although this application has been described in detail with reference to the foregoing embodiments, those skilled in the art should understand that modifications can still be made to the technical solutions described in the foregoing embodiments, or equivalent substitutions can be made to some of the technical features. Such modifications or substitutions do not cause the essence of the corresponding technical solutions to deviate from the spirit and scope of the technical solutions of the embodiments of this application, and should all be included within the protection scope of this application.
Claims
1. A method for determining a loop threshold, characterized in that, include: Linear regression was performed on the fluorescence values from the Nth to the Mth cycles of the amplification curve to be processed to obtain the baseline corresponding to the amplification curve; Both N and M are positive integers, and M is greater than N; Determine the residual standard error of the fluorescence values from the Nth cycle to the Mth cycle; The residual standard error is used to describe the average offset between the baseline and each cycle point from the Nth cycle to the Mth cycle; The threshold line corresponding to the amplification curve is determined based on the baseline and the residual standard error. The cycle threshold is determined based on the threshold line and the amplification curve; The cycle threshold refers to the number of cycles required for the fluorescence value to reach a preset threshold in a fluorescent polymerase chain reaction.
2. The determination method according to claim 1, characterized in that, N is 3 and M is 15.
3. The determination method according to claim 1, characterized in that, The determination of the threshold line based on the baseline and the residual standard error includes: Obtain the first slope and first intercept of the baseline; The second slope of the threshold line is determined based on the first slope; The second intercept of the threshold line is determined based on the first intercept and the residual standard error. The threshold line is determined based on the second slope and the second intercept.
4. The determination method according to claim 3, characterized in that, Determining the second slope of the threshold line based on the first slope includes: The first slope is determined as the second slope of the threshold line.
5. The determination method according to claim 3, characterized in that, The determination of the second intercept of the threshold line based on the first intercept and the residual standard error includes: The sum of the first intercept and the residual standard error of a preset multiple is determined as the second intercept of the threshold line.
6. The determination method according to claim 5, characterized in that, The preset multiple is greater than or equal to 10 and less than or equal to 30.
7. The determination method according to claim 1, wherein the amplification curve consists of multiple cycle points, and each cycle point is configured with a first cycle number corresponding to the cycle point and the fluorescence value, characterized in that, Determining the cycle threshold based on the threshold line and the amplification curve includes: Determine multiple equally spaced second cycle numbers between each pair of adjacent first cycle numbers; Determine the first comparison value corresponding to each of the second cycle numbers from the amplification curves; Determine the second comparison value for each of the second cycle numbers from the threshold lines; For any given second cycle number, if the absolute value of the difference between the first comparison value and the second comparison value corresponding to the second cycle number is less than or equal to a preset threshold, then the second cycle number is determined as the cycle threshold.
8. An electronic device, characterized in that, include: The first determining unit is used to perform linear regression processing on the fluorescence values of the Nth to Mth cycles in the amplification curve to be processed, so as to obtain the baseline corresponding to the amplification curve. Both N and M are positive integers, and M is greater than N; The second determining unit is used to determine the residual standard error of the fluorescence values from the Nth cycle to the Mth cycle in the amplification curve; The residual standard error is used to describe the average offset between the baseline and each cycle point from the Nth cycle to the Mth cycle; The third determining unit is used to determine the threshold line corresponding to the amplification curve based on the baseline and the residual standard error. The fourth determining unit is used to determine the cycle threshold based on the threshold line and the amplification curve; The cycle threshold refers to the number of cycles required for the fluorescence value to reach a preset threshold in a fluorescent polymerase chain reaction.
9. An electronic device comprising a memory, a processor, and a computer program stored in the memory and executable on the processor, characterized in that, When the processor executes the computer program, it implements each step of the method for determining the loop threshold as described in any one of claims 1 to 7.
10. A computer-readable storage medium storing a computer program, characterized in that, When the computer program is executed by a processor, it implements the steps in the method for determining the loop threshold as described in any one of claims 1 to 7.