QPCR (quantitative polymerase chain reaction) curve background phase detection method, qPCR curve background phase detection device and electronic equipment

Abstract

The invention discloses a qPCR curve background phase detection method and device and electronic equipment, and relates to the technical field of biological detection and the technical field of computers, and the method comprises the following steps: obtaining curve data of a to-be-detected qPCR curve; according to a plurality of background period cycles included in the current background period, calculating an average value of fluorescence values of the plurality of background period cycles; calculating the deviation degree of the fluorescence value of the last background period cycle of the current background period; determining whether the deviation degree meets a preset deviation degree judgment standard or not, wherein the deviation degree judgment standard is determined based on curve characteristics of a background period and an index growth period of a standard qPCR curve; if the preset deviation degree judgment standard is met, the background period of the qPCR curve is determined based on the current background period, and if the preset deviation degree judgment standard is not met, the current background period is adjusted to obtain the updated current background period, and the step of calculating the average value is executed again. By adopting the scheme, the accuracy of determining the background period of the qPCR curve and the detection efficiency are improved.

Description

Technical Field

[0001] The present application relates to the technical fields of biological detection and computer technology, and particularly relates to a method, device and electronic device for detecting the background period of a qPCR curve. Background Art

[0002] qPCR, that is, quantitative fluorescence PCR, is a method that utilizes the change of fluorescence signals to detect the change in the amount of amplification products in each cycle of PCR amplification in real time and perform quantitative analysis. As Figure 1 shown in the qPCR curve (which can also be called the qPCR amplification curve), it reflects the change of the corresponding fluorescence signals as the qPCR reaction proceeds. In the qPCR curve, the horizontal axis data is the cycle number. During the qPCR detection process, from the reagent temperature rising to a preset higher value to dropping to a preset lower value is one cycle. When the reagent temperature drops to the preset lower value, the fluorescence value of the reagent is collected as the fluorescence value of this cycle, that is, the vertical axis data in the qPCR curve.

[0003] As Figure 1 shown, the standard qPCR amplification curve presents an "S" shape with a characteristic form: first is the background signal (i.e., the background period, also known as the baseline period), and then there are three growth stages (exponential growth period, linear growth period, and plateau period).

[0004] When performing quantitative analysis on the qPCR curve, it is first necessary to determine the background period. Currently, there is no unified automatic method for determining the background period in the existing qPCR curve quantitative analysis technology, and the existing technologies have different performances in terms of detection speed, detection scenarios, and whether human assistance is required.

[0005] Currently, it is often the case that the 3rd - 15th cycles of the qPCR curve are determined as the range of the background period according to experience. However, in actual applications, this empirical value is often not accurate enough, or if it is determined by manual observation, the efficiency is relatively low.

[0006] Therefore, there are problems in the prior art that the determined background period of the qPCR curve is not accurate enough and the efficiency is relatively low. Summary of the Invention

[0007] Embodiments of the present application provide a method, device and electronic device for detecting the background period of a qPCR curve to solve the problems in the prior art that the determined background period of the qPCR curve is not accurate enough and the efficiency is relatively low.

[0008] Embodiments of the present application provide a method for detecting the background period of a qPCR curve, including:

[0009] Obtain the curve data of the qPCR curve to be detected, where the curve data includes multiple consecutive cycles included in the qPCR curve and the fluorescence value of each cycle;

[0010] Calculate the average value of the fluorescence values of the multiple background period cycles included in the current background period. The multiple background period cycles included in the initial background period are preset;

[0011] Based on the average value, calculate the deviation degree of the fluorescence value of the last background period cycle of the current background period;

[0012] Determine whether the deviation degree meets the preset deviation degree judgment criterion, where the deviation degree judgment criterion is determined based on the curve characteristics of the background period and the exponential growth period of the standard qPCR curve;

[0013] If the deviation degree meets the preset deviation degree judgment criterion, determine the background period of the qPCR curve based on the current background period. If the deviation degree does not meet the preset deviation degree judgment criterion, adjust the current background period to obtain the updated current background period, and return to execute the step of calculating the average value of the fluorescence values of the multiple background period cycles included in the current background period.

[0014] Further, the calculating the deviation degree of the fluorescence value of the last background period cycle of the current background period based on the average value includes:

[0015] Calculate the absolute value of the difference between the fluorescence value of the last background period cycle of the current background period and the average value as the deviation degree of the fluorescence value of the last background period cycle of the current background period.

[0016] Further, the determining whether the deviation degree meets the preset deviation degree judgment criterion includes:

[0017] Determine whether the deviation degree is within the preset deviation degree threshold range;

[0018] If it is within the preset deviation degree threshold range, determine that the deviation degree meets the preset deviation degree judgment criterion;

[0019] If it is not within the preset deviation degree threshold range, determine that the deviation degree does not meet the preset deviation degree judgment criterion.

[0020] Further, before the determining whether the deviation degree is within the preset deviation degree threshold range, it further includes:

[0021] Calculate the variance of the fluorescence values of the multiple background period cycles included in the current background period;

[0022] The preset deviation threshold range is from the variance of the first preset multiple to the variance of the second preset multiple, and the first preset multiple is less than the second preset multiple.

[0023] Further, if the deviation meets the preset deviation judgment criterion, based on the current background period, determining the background period of the qPCR curve includes:

[0024] If the deviation meets the preset deviation judgment criterion, determining the current background period as the background period of the qPCR curve;

[0025] If the deviation does not meet the preset deviation judgment criterion, adjusting the current background period to obtain an updated current background period, includes:

[0026] If the deviation is greater than the variance of the second preset multiple, excluding the last background period cycle from the multiple background period cycles included in the current background period to obtain an updated current background period;

[0027] If the deviation is less than the variance of the first preset multiple, adding the next cycle to the multiple background period cycles included in the current background period to obtain an updated current background period.

[0028] Further, determining whether the deviation meets the preset deviation judgment criterion includes:

[0029] Determining whether the deviation is not greater than the preset deviation threshold;

[0030] If the deviation meets the preset deviation judgment criterion, based on the current background period, determining the background period of the qPCR curve, if the deviation does not meet the preset deviation judgment criterion, adjusting the current background period to obtain an updated current background period, includes:

[0031] If the deviation is greater than the preset deviation threshold and the deviation calculated for the previous current background period is not greater than the preset deviation threshold, determining the previous current background period as the background period of the qPCR curve;

[0032] If the deviation is greater than the preset deviation threshold and the deviation calculated for the previous current background period is greater than the preset deviation threshold, excluding the last background period cycle from the multiple background period cycles included in the current background period to obtain an updated current background period;

[0033] If the deviation is not greater than the preset deviation threshold and the deviation calculated for the previous current background period is greater than the preset deviation threshold, determining the current background period as the background period of the qPCR curve;

[0034] If the deviation degree is not greater than the preset deviation degree threshold, and the deviation degree calculated for the previous current background period is not greater than the preset deviation degree threshold, in the multiple background period loops included in the current background period, add the next loop to obtain an updated current background period.

[0035] Further, before determining whether the deviation degree is within the preset deviation degree threshold range, it further includes:

[0036] Calculate the variance of the fluorescence values of the multiple background period loops according to the multiple background period loops included in the current background period;

[0037] [[ID=1-1]]The preset deviation degree threshold is the variance of the third preset multiple.

[0038] Further, the third preset multiple is a value between 1.5 times and 2 times.

[0039] The embodiment of the present application further provides a qPCR curve background period detection device, including:

[0040] A data acquisition module, configured to acquire curve data of a qPCR curve to be detected, where the curve data includes a plurality of consecutive loops included in the qPCR curve and the fluorescence value of each loop;

[0041] An average value calculation module, configured to calculate the average value of the fluorescence values of the multiple background period loops according to the multiple background period loops included in the current background period, and the multiple background period loops included in the initial background period are preset;

[0042] A deviation degree calculation module, configured to calculate the deviation degree of the fluorescence value of the last background period loop of the current background period based on the average value;

[0043] A deviation degree judgment module, configured to determine whether the deviation degree meets a preset deviation degree judgment criterion, and the deviation degree judgment criterion is determined based on the curve characteristics of the background period and the exponential growth period of the standard qPCR curve;

[0044] A background period adjustment module, configured to, if the deviation degree meets the preset deviation degree judgment criterion, determine the background period of the qPCR curve based on the current background period, and if the deviation degree does not meet the preset deviation degree judgment criterion, adjust the current background period to obtain an updated current background period, and return to the average value calculation module to execute the step of calculating the average value of the fluorescence values of the multiple background period loops according to the multiple background period loops included in the current background period.

[0045] Further, the deviation calculation module is specifically configured to calculate the absolute value of the difference between the fluorescence value of the last background period cycle of the current background period and the average value as the deviation of the fluorescence value of the last background period cycle of the current background period.

[0046] Further, the deviation determination module is specifically configured to determine whether the deviation is within a preset deviation threshold range; if it is within the preset deviation threshold range, it is determined that the deviation meets the preset deviation determination criterion; if it is not within the preset deviation threshold range, it is determined that the deviation does not meet the preset deviation determination criterion.

[0047] Further, the average value calculation module is further configured to calculate the variance of the fluorescence values of the multiple background period cycles included in the current background period; the preset deviation threshold range is from the variance of the first preset multiple to the variance of the second preset multiple, and the first preset multiple is less than the second preset multiple.

[0048] Further, the background period adjustment module is specifically configured to, if the deviation meets the preset deviation determination criterion, determine that the current background period is the background period of the qPCR curve; if the deviation is greater than the variance of the second preset multiple, exclude the last background period cycle from the multiple background period cycles included in the current background period to obtain an updated current background period; if the deviation is less than the variance of the first preset multiple, add the next cycle to the multiple background period cycles included in the current background period to obtain an updated current background period.

[0049] Further, the deviation determination module is specifically configured to determine whether the deviation is not greater than a preset deviation threshold.

[0050] The background period adjustment module is specifically configured to determine the previous current background period as the background period of the qPCR curve if the deviation degree is greater than the preset deviation degree threshold and the deviation degree calculated for the previous current background period is not greater than the preset deviation degree threshold; if the deviation degree is greater than the preset deviation degree threshold and the deviation degree calculated for the previous current background period is greater than the preset deviation degree threshold, remove the last background period loop from the multiple background period loops included in the current background period to obtain an updated current background period; if the deviation degree is not greater than the preset deviation degree threshold and the deviation degree calculated for the previous current background period is greater than the preset deviation degree threshold, determine the current background period as the background period of the qPCR curve; if the deviation degree is not greater than the preset deviation degree threshold and the deviation degree calculated for the previous current background period is not greater than the preset deviation degree threshold, add the next loop to the multiple background period loops included in the current background period to obtain an updated current background period.

[0051] Further, the average value calculation module is further configured to calculate the variance of the fluorescence values of the multiple background period loops according to the multiple background period loops included in the current background period; the preset deviation degree threshold is three times the variance.

[0052] Further, the third preset multiple is a value between 1.5 times and 2 times.

[0053] An embodiment of the present application further provides an electronic device, including a processor and a machine-readable storage medium, where the machine-readable storage medium stores machine-executable instructions that can be executed by the processor, and the processor is prompted by the machine-executable instructions to implement the qPCR curve background period detection method described in any one of the above.

[0054] An embodiment of the present application further provides a computer-readable storage medium, characterized in that the computer-readable storage medium stores a computer program, and when the computer program is executed by a processor, the qPCR curve background period detection method described in any one of the above is implemented.

[0055] An embodiment of the present application further provides a computer program product including instructions, and when it runs on a computer, it causes the computer to execute the qPCR curve background period detection method described in any one of the above.

[0056] The beneficial effects of the present application include:

[0057] In the method provided by the embodiments of the present application, an initial background period is preset, and for the current background period, the average value of the fluorescence values of multiple background period cycles included therein is calculated, and the deviation degree of the fluorescence value of the last background period cycle is calculated. Based on whether the deviation degree meets the preset deviation degree judgment criterion, the current background period is adjusted, and the background period of the qPCR curve is determined. Since the deviation degree judgment criterion is determined based on the curve characteristics of the background period and the exponential growth period of the standard qPCR curve, compared with the prior art, based on this deviation degree judgment criterion, a more accurate background period can be detected, and automatic detection of the background period can be realized, improving the detection efficiency.

[0058] Other features and advantages of the present application will be described in the following specification, and in part will be obvious from the specification, or will be understood by implementing the present application. The objectives and other advantages of the present application can be realized and obtained by the structures specifically pointed out in the written specification, claims, and drawings BRIEF DESCRIPTION OF THE DRAWINGS

[0059] The drawings are used to provide a further understanding of the present application, and constitute a part of the specification. Together with the embodiments of the present application, they are used to explain the present application, and do not constitute a limitation to the present application. In the drawings:

[0060] Figure 1 It is a schematic diagram of a standard qPCR curve;

[0061] Figure 2 It is a flowchart of the qPCR curve background period detection method provided by the embodiments of the present application;

[0062] Figure 3 It is a flowchart of the qPCR curve background period detection method provided by Embodiment 1 of the present application;

[0063] Figure 4 It is a flowchart of the qPCR curve background period detection method provided by Embodiment 2 of the present application;

[0064] Figure 5 It is a schematic structural diagram of the qPCR curve background period detection device provided by the embodiments of the present application;

[0065] Figure 6 It is a schematic structural diagram of the electronic device provided by the embodiments of the present application. DETAILED DESCRIPTION OF THE EMBODIMENTS

[0066] In order to provide a solution for improving the accuracy of determining the background period of a qPCR curve and realizing the detection efficiency, the embodiments of the present application provide a method, a device and an electronic device for detecting the background period of a qPCR curve. The preferred embodiments of the present application will be described below with reference to the accompanying drawings. It should be understood that the preferred embodiments described herein are only used to illustrate and explain the present application and are not used to limit the present application. And without conflict, the embodiments in the present application and the features in the embodiments can be combined with each other.

[0067] The embodiments of the present application provide a method for detecting the background period of a qPCR curve. As Figure 2 shown, it includes:

[0068] Step 21, obtain the curve data of the qPCR curve to be detected, where the curve data includes a plurality of consecutive cycles included in the qPCR curve and the fluorescence value of each cycle;

[0069] Step 22, calculate the average value of the fluorescence values of the multiple background period cycles included in the current background period, and the multiple background period cycles included in the initial background period are preset;

[0070] Step 23, calculate the deviation degree of the fluorescence value of the last background period cycle of the current background period based on the average value;

[0071] Step 24, determine whether the deviation degree meets the preset deviation degree judgment criterion, and the deviation degree judgment criterion is determined based on the curve characteristics of the background period and the exponential growth period of the standard qPCR curve;

[0072] Step 25, if the deviation degree meets the preset deviation degree judgment criterion, determine the background period of the qPCR curve based on the current background period. If the deviation degree does not meet the preset deviation degree judgment criterion, adjust the current background period to obtain the updated current background period, and return to execute Step 22.

[0073] In the method for detecting the background period of the qPCR curve provided by the embodiments of the present application, since the deviation degree judgment criterion is determined based on the curve characteristics of the background period and the exponential growth period of the standard qPCR curve, compared with the prior art, based on this deviation degree judgment criterion, a more accurate background period can be detected, and the automatic detection of the background period can be realized, improving the detection efficiency.

[0074] The method and device provided by the present application will be described in detail below with specific embodiments in conjunction with the accompanying drawings.

[0075] Embodiment 1:

[0076] The embodiment 1 of the present application provides a method for detecting the background period of a qPCR curve. As Figure 3 shown, it includes:

[0077] Step 31: Obtain the curve data of the qPCR curve to be detected. The curve data includes a plurality of consecutive cycles included in the qPCR curve and the fluorescence value of each cycle.

[0078] Step 32: Calculate the average value of the fluorescence values of the multiple background period cycles included in the current background period. The multiple background period cycles included in the initial background period are preset.

[0079] In Embodiment 1 of the present application, the multiple cycles included in the initial background period can be preset as the 3rd - 15th cycles. For the convenience of description, the cycles included in the background period are referred to as background period cycles.

[0080] When this step is first executed, the current background period is the initial background period, that is, calculate the average value of the fluorescence values of the multiple background period cycles included in the initial background period.

[0081] In Embodiment 1 of the present application, the current background period does not mean that the background period of the qPCR curve is changing, but during the process of detecting the background period of the qPCR curve, that is, during the continuous adjustment of the background period, as the background period that needs to be judged currently. For the convenience of description, it is referred to as the current background period.

[0082] Step 33: Based on the average value, calculate the deviation degree of the fluorescence value of the last background period cycle of the current background period.

[0083] In this step, the absolute value of the difference between the fluorescence value of the last background period cycle of the current background period and the average value can be calculated as the deviation degree of the fluorescence value of the last background period cycle of the current background period. That is, it can be calculated using the following formula:

[0084] Flat(i) = |FA(i) - AVG|;

[0085] Wherein, i represents the last background period cycle of the current background period, FA(i) represents the fluorescence value of the last background period cycle of the current background period, AVA represents the average value of the fluorescence values of the multiple background period cycles included in the current background period, and Flat(i) represents the deviation degree of the fluorescence value of the last background period cycle of the current background period.

[0086] Step 34: Calculate the variance of the fluorescence values of the multiple background period cycles included in the current background period.

[0087] There is no strict sequence between this Step 34 and the above Steps 32 - 33.

[0088] Step 35: Determine whether the above deviation degree is within the preset deviation degree threshold range.

[0089] If within the preset deviation threshold range, it is determined that the deviation meets the preset deviation judgment criterion, execute step 36;

[0090] If not within the preset deviation threshold range, it is determined that the deviation does not meet the preset deviation judgment criterion, execute step 37.

[0091] In Embodiment 1 of the present application, the preset deviation threshold range is determined based on the curve characteristics of the background period and the exponential growth period of the standard qPCR curve.

[0092] From Figure 1 As can be seen from the ranges of the background period and the exponential growth period of the shown standard qPCR curve, from the background period to the exponential growth period, the fluorescence value will have an obvious increase.

[0093] Furthermore, in Embodiment 1 of the present application, the preset deviation threshold range can be related to the variance of the fluorescence values of the multiple background period cycles included in the current background period determined in step 34 above.

[0094] For example, the preset deviation threshold range is from the first preset multiple of the variance to the second preset multiple of the variance, and the first preset multiple is less than the second preset multiple.

[0095] In practical applications, the first preset multiple and the second preset multiple can be set to appropriate values based on the amplification performance of the instrument. If the amplification performance of the instrument is relatively less sensitive, the values of the first preset multiple and the second preset multiple can be set relatively large. If the amplification performance of the instrument is relatively more sensitive, the values of the first preset multiple and the second preset multiple can be set relatively small.

[0096] Specifically, in Embodiment 1 of the present application, the preset deviation threshold range can be set to 1.9 times the variance to 2 times the variance, or, set to 1.8 times the variance to 1.9 times the variance, or, set to 1.7 times the variance to 1.8 times the variance, or, set to 1.6 times the variance to 1.7 times the variance, or, set to 1.5 times the variance to 1.6 times the variance.

[0097] Step 36: If the deviation meets the preset deviation judgment criterion, determine the current background period as the background period of the qPCR curve.

[0098] According to step 35 above, if the deviation is within the preset deviation threshold range, it means that the last background period cycle of the current background period is at the junction of the background period and the exponential growth period. Therefore, the current background period can be determined as the background period of the qPCR curve.

[0099] Step 37: If the deviation does not meet the preset deviation judgment criterion, adjust the current background period to obtain an updated current background period, and return to execute the above Step 32 to continue determining whether the conditions for being a background period are met for the updated current background period.

[0100] In this step, if the deviation is greater than the variance of the second preset multiple, it indicates that the last cycle of the current background period is still within the exponential growth period range. Therefore, remove the last background period cycle from the multiple background period cycles included in the current background period to obtain an updated current background period. For example, if the multiple cycles included in the current background period are cycles 3 - 15, then remove cycle 15, and the multiple cycles included in the updated current background period change to cycles 3 - 14;

[0101] If the deviation is less than the variance of the first preset multiple, it indicates that the last cycle of the current background period has not yet reached the junction between the background period and the exponential growth period. Therefore, add the next cycle to the multiple background period cycles included in the current background period to obtain an updated current background period. For example, if the multiple cycles included in the current background period are cycles 3 - 15, then add the next cycle, i.e., cycle 16, and the multiple cycles included in the updated current background period change to cycles 3 - 16.

[0102] Using the qPCR curve background period detection method provided in Embodiment 1 of the present application above Figure 3 shown, the preset deviation judgment criterion uses a preset deviation threshold range, and this preset deviation threshold range is related to the variance of the fluorescence values of the multiple background period cycles included in the current background period and is determined based on the curve characteristics of the background period and the exponential growth period of the standard qPCR curve. Therefore, the cycle located at the junction between the background period and the exponential growth period can be accurately found from multiple cycles as the last cycle of the background period to be detected for this qPCR curve. Compared with the prior art, a more accurate background period can be detected, and automatic detection of the background period can be achieved, improving the detection efficiency.

[0103] Embodiment 2:

[0104] The qPCR curve background period detection method provided in Embodiment 2 of the present application, as Figure 4 shown, includes:

[0105] Steps 401 - 404 are respectively the same as Steps 31 - 34 in the above Embodiment 1 and will not be repeated here.

[0106] Step 405: Determine whether the above deviation is not greater than the preset deviation threshold.

[0107] If it is greater than the preset deviation threshold, execute Step 406.

[0108] If it is not greater than the preset deviation threshold, step 409 is executed.

[0109] In Embodiment 2 of the present application, the preset deviation threshold is determined based on the curve characteristics of the background period and the exponential growth period of the standard qPCR curve.

[0110] From Figure 1 As can be seen from the ranges of the background period and the exponential growth period of the shown standard qPCR curve, from the background period to the exponential growth period, the fluorescence value will have an obvious increase.

[0111] Further, in Embodiment 2 of the present application, the preset deviation threshold range can be related to the variance of the fluorescence values of multiple background period cycles included in the current background period determined in step 34 above.

[0112] For example, the preset deviation threshold is the third preset multiple of this variance.

[0113] In practical applications, for the third preset multiple, an appropriate value can be set based on the amplification performance of the instrument. If the amplification performance of the instrument is less sensitive, the value of the third preset multiple can be set relatively large. If the amplification performance of the instrument is more sensitive, the value of the third preset multiple can be set relatively small.

[0114] Specifically, in Embodiment 2 of the present application, the preset deviation threshold can be set to a value between 1.5 times the variance and 2 times the variance.

[0115] Step 406: Determine whether the deviation calculated for the previous current background period is not greater than the preset deviation threshold.

[0116] If it is not greater than the preset deviation threshold, step 407 is executed.

[0117] If it is greater than the preset deviation threshold, step 408 is executed.

[0118] Step 407: If the deviation calculated for the current background period is greater than the preset deviation threshold, and the deviation calculated for the previous current background period is not greater than the preset deviation threshold, it means that the last cycle of the current background period is just the first cycle from the background period to the exponential growth period. Therefore, the previous current background period can be determined as the background period of the qPCR curve.

[0119] Step 408: If the deviation calculated for the current background period is greater than the preset deviation threshold, and the deviation calculated for the previous current background period is greater than the preset deviation threshold, it indicates that the last cycle of the current background period is still within the exponential growth period range. Therefore, eliminate the last background period cycle from the multiple background period cycles included in the current background period to obtain the updated current background period, and return to execute the above Step 402 to continue determining whether the updated current background period meets the conditions for being a background period.

[0120] For example, if the multiple cycles included in the current background period are the 3rd - 15th cycles, then eliminate the 15th cycle, and the multiple cycles included in the updated current background period change to the 3rd - 14th cycles.

[0121] 409. Determine whether the deviation calculated for the previous current background period is not greater than the preset deviation threshold.

[0122] If it is not greater than the preset deviation threshold, execute Step 410.

[0123] If it is greater than the preset deviation threshold, execute Step 411.

[0124] Step 410: If the deviation calculated for the current background period is not greater than the preset deviation threshold, and the deviation calculated for the previous current background period is greater than the preset deviation threshold, it indicates that the last cycle of the current background period is exactly the last cycle of the background period. Therefore, determine the current background period as the background period of the qPCR curve.

[0125] Step 411: If the deviation calculated for the current background period is not greater than the preset deviation threshold, and the deviation calculated for the previous current background period is not greater than the preset deviation threshold, it indicates that the last cycle of the current background period may not yet be at the junction of the background period and the exponential growth period. Therefore, add the next cycle to the multiple background period cycles included in the current background period to obtain the updated current background period, and return to execute the above Step 402 to continue determining whether the updated current background period meets the conditions for being a background period.

[0126] For example, if the multiple cycles included in the current background period are the 3rd - 15th cycles, then add the next cycle, that is, the 16th cycle, and the multiple cycles included in the updated current background period change to the 3rd - 16th cycles.

[0127] The above Figure 4The process shown is essentially to find a cycle from multiple cycles included in the qPCR curve to be detected, for which the calculated deviation degree is not greater than a preset deviation degree threshold, and the deviation degree calculated for the next cycle of this cycle is greater than the preset deviation degree threshold, that is, taking this cycle as the last cycle among the multiple cycles included in the background period, so as to determine the background period of the qPCR curve.

[0128] Using the above-mentioned Figure 4 qPCR curve background period detection method provided in Embodiment 2 of the present application, the preset deviation degree judgment criterion uses a preset deviation degree threshold, and this preset deviation degree threshold is related to the variance of the fluorescence values of the multiple background period cycles included in the current background period, and is determined based on the curve characteristics of the background period and the exponential growth period of the standard qPCR curve. Therefore, it is possible to accurately find the cycle at the junction of the background period and the exponential growth period from multiple cycles as the last cycle of the background period of the qPCR curve to be detected. Compared with the prior art, a more accurate background period can be detected, and automatic detection of the background period can be realized, improving the detection efficiency.

[0129] Based on the same inventive concept, according to the qPCR curve background period detection method provided in the above embodiments of the present application, correspondingly, another embodiment of the present application also provides a qPCR curve background period detection device, and its structural schematic diagram is as Figure 5 shown, specifically including:

[0130] A data acquisition module 51, configured to acquire curve data of the qPCR curve to be detected, where the curve data includes a plurality of consecutive cycles included in the qPCR curve, and the fluorescence value of each cycle;

[0131] An average value calculation module 52, configured to calculate the average value of the fluorescence values of the multiple background period cycles according to the multiple background period cycles included in the current background period, and the multiple background period cycles included in the initial background period are preset;

[0132] A deviation degree calculation module 53, configured to calculate the deviation degree of the fluorescence value of the last background period cycle of the current background period based on the average value;

[0133] A deviation degree judgment module 54, configured to determine whether the deviation degree meets a preset deviation degree judgment criterion, and the deviation degree judgment criterion is determined based on the curve characteristics of the background period and the exponential growth period of the standard qPCR curve;

[0134] The background period adjustment module 55 is used to determine the background period of the qPCR curve based on the current background period if the deviation meets the preset deviation judgment criterion. If the deviation does not meet the preset deviation judgment criterion, the current background period is adjusted to obtain an updated current background period, and the average value calculation module is returned to execute the step of calculating the average value of the fluorescence values of the multiple background period cycles included in the current background period in a loop.

[0135] Further, the deviation calculation module 53 is specifically configured to calculate the absolute value of the difference between the fluorescence value of the last background period cycle of the current background period and the average value as the deviation of the fluorescence value of the last background period cycle of the current background period.

[0136] Further, the deviation judgment module 54 is specifically configured to determine whether the deviation is within a preset deviation threshold range; if it is within the preset deviation threshold range, it is determined that the deviation meets the preset deviation judgment criterion; if it is not within the preset deviation threshold range, it is determined that the deviation does not meet the preset deviation judgment criterion.

[0137] Further, the average value calculation module 52 is further configured to calculate the variance of the fluorescence values of the multiple background period cycles included in the current background period; the preset deviation threshold range is from the variance of the first preset multiple to the variance of the second preset multiple, and the first preset multiple is less than the second preset multiple.

[0138] Further, the background period adjustment module 55 is specifically configured to determine that the current background period is the background period of the qPCR curve if the deviation meets the preset deviation judgment criterion; if the deviation is greater than the variance of the second preset multiple, the last background period cycle is removed from the multiple background period cycles included in the current background period to obtain an updated current background period; if the deviation is less than the variance of the first preset multiple, the next cycle is added to the multiple background period cycles included in the current background period to obtain an updated current background period.

[0139] Further, the deviation judgment module 54 is specifically configured to determine whether the deviation is not greater than a preset deviation threshold;

[0140] The background period adjustment module 55 is specifically configured to determine the previous current background period as the background period of the qPCR curve if the deviation degree is greater than the preset deviation degree threshold and the deviation degree calculated for the previous current background period is not greater than the preset deviation degree threshold; if the deviation degree is greater than the preset deviation degree threshold and the deviation degree calculated for the previous current background period is greater than the preset deviation degree threshold, remove the last background period cycle from the multiple background period cycles included in the current background period to obtain an updated current background period; if the deviation degree is not greater than the preset deviation degree threshold and the deviation degree calculated for the previous current background period is greater than the preset deviation degree threshold, determine the current background period as the background period of the qPCR curve; if the deviation degree is not greater than the preset deviation degree threshold and the deviation degree calculated for the previous current background period is not greater than the preset deviation degree threshold, add the next cycle to the multiple background period cycles included in the current background period to obtain an updated current background period.

[0141] Further, the average value calculation module 52 is further configured to calculate the variance of the fluorescence values of the multiple background period cycles according to the multiple background period cycles included in the current background period; the preset deviation degree threshold is the third preset multiple of the variance.

[0142] Further, the third preset multiple is a value between 1.5 times and 2 times.

[0143] The functions of the above modules can correspond to Figures 2 to 4 the corresponding processing steps in the shown process, which will not be elaborated here.

[0144] The qPCR curve background period detection device provided by the embodiments of the present application can be implemented by a computer program. Those skilled in the art should be able to understand that the above module division method is only one of many module division methods. If divided into other modules or not divided into modules, as long as the qPCR curve background period detection device has the above functions, it should be within the protection scope of the present application.

[0145] The embodiments of the present application also provide an electronic device, as Figure 6 shown, including a processor 61 and a machine-readable storage medium 62. The machine-readable storage medium 62 stores machine-executable instructions that can be executed by the processor 61. The processor 61 is prompted by the machine-executable instructions to implement the qPCR curve background period detection method described in any one of the above.

[0146] The embodiments of the present application also provide a computer-readable storage medium, characterized in that the computer-readable storage medium stores a computer program, and when the computer program is executed by a processor, it implements the qPCR curve background period detection method described in any one of the above.

[0147] An embodiment of the present application also provides a computer program product including instructions, which, when running on a computer, causes the computer to execute the qPCR curve background period detection method described in any one of the above.

[0148] The machine-readable storage medium in the above electronic device may include a random access memory (RAM), or may also include a non-volatile memory (NVM), such as at least one disk memory. Optionally, the memory may also be at least one storage device located far from the aforementioned processor.

[0149] The above-mentioned processor may be a general-purpose processor, including a central processing unit (CPU), a network processor (NP), etc.; it may also be a digital signal processor (DSP), an application specific integrated circuit (ASIC), a field-programmable gate array (FPGA), or other programmable logic devices, discrete gate or transistor logic devices, discrete hardware components.

[0150] Each embodiment in this specification is described in a related manner. The same or similar parts between the embodiments can be referred to each other, and the differences between each embodiment and other embodiments are emphasized. In particular, for the device, electronic device, computer-readable storage medium, and computer program product embodiments, since they are basically similar to the method embodiments, the description is relatively simple, and the relevant parts can be referred to the description of the method embodiments.

[0151] It should be noted that in this article, relational terms such as first and second are only used to distinguish one entity or operation from another entity or operation, and do not necessarily require or imply any actual relationship or order between these entities or operations. Moreover, the term "comprising", "including" or any other variant thereof is intended to cover non-exclusive inclusion, so that a process, method, article or device including a series of elements not only includes those elements, but also includes other elements not expressly listed, or also includes elements inherent to such process, method, article or device. Without further limitation, an element defined by the statement "including one..." does not exclude the existence of another identical element in the process, method, article or device including the element.

[0152] This application is described with reference to the flowcharts and / or block diagrams of methods, apparatuses (systems), and computer program products according to embodiments of the present application. It should be understood that each flow and / or block in the flowcharts and / or block diagrams, as well as the combination of flows and / or blocks in the flowcharts and / or block diagrams, can be implemented by computer program instructions. These computer program instructions can be provided to the processor of a general-purpose computer, a special-purpose computer, an embedded processor, or other programmable data processing devices to generate a machine, such that the instructions executed by the processor of the computer or other programmable data processing devices generate means for implementing the functions specified in one flow Figure 1 one flow or multiple flows and / or blocks Figure 1 or means for implementing the functions specified in one block or multiple blocks.

[0153] These computer program instructions can also be stored in a computer-readable memory that can direct a computer or other programmable data processing device to work in a specific manner, such that the instructions stored in the computer-readable memory generate a manufactured article including instruction means that implement the functions specified in one flow Figure 1 one flow or multiple flows and / or blocks Figure 1 or means for implementing the functions specified in one block or multiple blocks.

[0154] These computer program instructions can also be loaded onto a computer or other programmable data processing device, such that a series of operation steps are executed on the computer or other programmable device to generate a computer-implemented process, so that the instructions executed on the computer or other programmable device provide steps for implementing the functions specified in one flow Figure 1 one flow or multiple flows and / or blocks Figure 1 or means for implementing the functions specified in one block or multiple blocks.

[0155] Obviously, those skilled in the art can make various changes and modifications to this application without departing from the spirit and scope of this application. Thus, if these modifications and variations of this application fall within the scope of the claims of this application and their equivalent technologies, this application is also intended to include these changes and modifications.

Claims

1. A method for detecting the background period of a qPCR curve, characterized in that, Including: Obtain the curve data of the qPCR curve to be detected, where the curve data includes a plurality of consecutive cycles included in the qPCR curve, and the fluorescence value of each cycle; Calculate the average value of the fluorescence values of the multiple background period cycles included in the current background period. The multiple background period cycles included in the initial background period are preset; Based on the average value, calculate the deviation degree of the fluorescence value of the last background period cycle of the current background period; Determine whether the deviation degree meets a preset deviation degree judgment criterion, where the deviation degree judgment criterion is determined based on the curve characteristics of the background period and the exponential growth period of the standard qPCR curve; If the deviation degree meets the preset deviation degree judgment criterion, determine the background period of the qPCR curve based on the current background period. If the deviation degree does not meet the preset deviation degree judgment criterion, adjust the current background period to obtain an updated current background period, and return to execute the step of calculating the average value of the fluorescence values of the multiple background period cycles included in the current background period.

2. The method according to claim 1, wherein The calculating the deviation degree of the fluorescence value of the last background period cycle of the current background period based on the average value includes: Calculate the absolute value of the difference between the fluorescence value of the last background period cycle of the current background period and the average value as the deviation degree of the fluorescence value of the last background period cycle of the current background period.

3. The method according to claim 1, characterized in that, The determining whether the deviation degree meets the preset deviation degree judgment criterion includes: Determine whether the deviation degree is within a preset deviation degree threshold range; If it is within the preset deviation degree threshold range, determine that the deviation degree meets the preset deviation degree judgment criterion; If it is not within the preset deviation degree threshold range, determine that the deviation degree does not meet the preset deviation degree judgment criterion.

4. The method according to claim 3, wherein Before the determining whether the deviation degree is within the preset deviation degree threshold range, it further includes: Calculate the variance of the fluorescence values of the multiple background period cycles included in the current background period; The preset deviation degree threshold range is from the first preset multiple of the variance to the second preset multiple of the variance, and the first preset multiple is less than the second preset multiple.

5. The method according to claim 4, wherein The if the deviation degree meets the preset deviation degree judgment criterion, determining the background period of the qPCR curve based on the current background period includes: If the deviation degree meets the preset deviation degree judgment criterion, determine the current background period as the background period of the qPCR curve; The if the deviation degree does not meet the preset deviation degree judgment criterion, adjusting the current background period to obtain an updated current background period includes: If the deviation degree is greater than the second preset multiple of the variance, exclude the last background period cycle from the multiple background period cycles included in the current background period to obtain an updated current background period; If the deviation degree is less than the first preset multiple of the variance, add the next cycle to the multiple background period cycles included in the current background period to obtain an updated current background period.

6. The method according to claim 1, wherein The determining whether the deviation degree meets the preset deviation degree judgment criterion includes: Determine whether the deviation degree is not greater than a preset deviation degree threshold; If the deviation degree meets the preset deviation degree judgment criterion, based on the current background period, determine the background period of the qPCR curve. If the deviation degree does not meet the preset deviation degree judgment criterion, adjust the current background period to obtain an updated current background period, including: If the deviation degree is greater than the preset deviation degree threshold and the deviation degree calculated for the previous current background period is not greater than the preset deviation degree threshold, determine the previous current background period as the background period of the qPCR curve; If the deviation degree is greater than the preset deviation degree threshold and the deviation degree calculated for the previous current background period is greater than the preset deviation degree threshold, remove the last background period cycle from the multiple background period cycles included in the current background period to obtain an updated current background period; If the deviation degree is not greater than the preset deviation degree threshold and the deviation degree calculated for the previous current background period is greater than the preset deviation degree threshold, determine the current background period as the background period of the qPCR curve; If the deviation degree is not greater than the preset deviation degree threshold and the deviation degree calculated for the previous current background period is not greater than the preset deviation degree threshold, add the next cycle to the multiple background period cycles included in the current background period to obtain an updated current background period.

7. The method according to claim 6, characterized in that, Before determining whether the deviation degree is within the preset deviation degree threshold range, it also includes: Calculate the variance of the fluorescence values of the multiple background period cycles included in the current background period according to the multiple background period cycles included in the current background period; The preset deviation degree threshold is three times the preset multiple of the variance.

8. The method according to claim 7, wherein The third preset multiple is a value between 1.5 times and 2 times.

9. A qPCR curve background period detection device, characterized in that, It includes: A data acquisition module for acquiring the curve data of the qPCR curve to be detected, where the curve data includes a plurality of consecutive cycles included in the qPCR curve and the fluorescence value of each cycle; An average value calculation module for calculating the average value of the fluorescence values of the multiple background period cycles according to the multiple background period cycles included in the current background period, and the multiple background period cycles included in the initial background period are preset; A deviation degree calculation module for calculating the deviation degree of the fluorescence value of the last background period cycle of the current background period based on the average value; A deviation degree judgment module for determining whether the deviation degree meets the preset deviation degree judgment criterion, and the deviation degree judgment criterion is determined based on the curve characteristics of the background period and the exponential growth period of the standard qPCR curve; A background period adjustment module for, if the deviation degree meets the preset deviation degree judgment criterion, determining the background period of the qPCR curve based on the current background period, if the deviation degree does not meet the preset deviation degree judgment criterion, adjusting the current background period to obtain an updated current background period, and returning to the average value calculation module to execute the step of calculating the average value of the fluorescence values of the multiple background period cycles included in the current background period.

10. The device according to claim 9, characterized in that, The deviation calculation module is specifically configured to calculate the absolute value of the difference between the fluorescence value of the last background period cycle of the current background period and the average value as the deviation of the fluorescence value of the last background period cycle of the current background period.

11. The device according to claim 9, characterized in that, The deviation judgment module is specifically configured to determine whether the deviation is within a preset deviation threshold range; if it is within the preset deviation threshold range, it is determined that the deviation meets the preset deviation judgment criteria; if it is not within the preset deviation threshold range, it is determined that the deviation does not meet the preset deviation judgment criteria.

12. The device according to claim 11, characterized in that, The average value calculation module is further configured to calculate the variance of the fluorescence values of the multiple background period cycles included in the current background period; the preset deviation threshold range is from the first preset multiple of the variance to the second preset multiple of the variance, and the first preset multiple is less than the second preset multiple.

13. The device according to claim 12, wherein The background period adjustment module is specifically configured to, if the deviation meets the preset deviation judgment criteria, determine that the current background period is the background period of the qPCR curve; if the deviation is greater than the second preset multiple of the variance, remove the last background period cycle from the multiple background period cycles included in the current background period to obtain an updated current background period; if the deviation is less than the first preset multiple of the variance, add the next cycle to the multiple background period cycles included in the current background period to obtain an updated current background period.

14. The device according to claim 9, characterized in that, The deviation judgment module is specifically configured to determine whether the deviation is not greater than a preset deviation threshold. The background period adjustment module is specifically configured to, if the deviation is greater than the preset deviation threshold and the deviation calculated for the previous current background period is not greater than the preset deviation threshold, determine that the previous current background period is the background period of the qPCR curve. If the deviation is greater than the preset deviation threshold and the deviation calculated for the previous current background period is greater than the preset deviation threshold, remove the last background period cycle from the multiple background period cycles included in the current background period to obtain an updated current background period; if the deviation is not greater than the preset deviation threshold and the deviation calculated for the previous current background period is greater than the preset deviation threshold, determine that the current background period is the background period of the qPCR curve; If the deviation is not greater than the preset deviation threshold and the deviation calculated for the previous current background period is not greater than the preset deviation threshold, add the next cycle to the multiple background period cycles included in the current background period to obtain an updated current background period.

15. The device according to claim 14, characterized in that, The average value calculation module is further configured to calculate the variance of the fluorescence values of the multiple background period cycles included in the current background period; the preset deviation threshold is the third preset multiple of the variance.

16. The device according to claim 15, wherein The third preset multiple is a value between 1.5 times and 2 times.

17. An electronic device, characterized in that, Comprising a processor and a machine-readable storage medium, the machine-readable storage medium stores machine-executable instructions that can be executed by the processor, and the processor is caused by the machine-executable instructions to: implement the method according to any one of claims 1-8.

18. A computer-readable storage medium, characterized in that, A computer program is stored in the computer-readable storage medium, and when the computer program is executed by a processor, the method according to any one of claims 1-8 is implemented.

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