A method and apparatus for detecting via events, an electronic device, and a storage medium

Abstract

The application discloses a kind of methods for detecting via events, device, electronic equipment and storage medium, method includes: the ion current between first cavity and second cavity is measured, obtain the ion current value of preset quantity;Wherein, first cavity is filled with base sequence;The preset quantity of ion current value is divided into multiple current value sets;Respectively, the extraction of peak value is carried out to multiple current value sets, determine multiple current peak values corresponding to multiple current value sets;Based on multiple current peak values and multiple current value sets respectively corresponding baseline current value, determine the current amplitude corresponding to multiple current peak values respectively;Based on the current amplitude corresponding to multiple current peak values respectively, determine detection threshold value;Based on the current amplitude corresponding to multiple current peak values respectively and detection threshold value, determine at least one via event of base sequence.Based on the scheme, the efficiency and accuracy of via event detection can be improved.

Description

Technical Field

[0001] This invention relates to the fields of bioinformatics and data analysis, and in particular to a method, apparatus, electronic device, and storage medium for detecting via events. Background Technology

[0002] Nanopore sequencing is an electrical signal-based sequencing technology that identifies sequences by capturing changes in current caused by different DNA sequences passing through nanopores. A nanopore event specifically refers to a significant change in current caused by the passage of a DNA sequence through a nanopore. Nanopore event detection refers to determining the time frame of a nanopore event by detecting abnormal changes in current. In the past decade or so, with continuous advancements in the technology, equipment, and materials for fabricating solid-state nanopores, research in this field has made rapid progress. However, the technology for using solid-state nanopores for gene sequencing is still imperfect, especially direct sequencing using solid-state nanopores, which remains a major challenge. This is mainly due to the difficulty in controlling the speed at which DNA passes through the nanopore and the low signal-to-noise ratio of the electrical signals output by solid-state nanopore sequencing devices.

[0003] Related technologies and via event detection methods mostly rely on setting thresholds for determination. This method is quite sensitive to parameter selection, resulting in low accuracy. On the other hand, the signals generated by via events need to be observed manually to determine the minimum amplitude, area, and other filtering conditions in the via event. The sequencing time is long, leading to slow via event detection efficiency. Summary of the Invention

[0004] This invention provides a method, apparatus, electronic device, and storage medium for detecting via events, which improves the efficiency and accuracy of via event detection.

[0005] The technical solution of this invention is implemented as follows:

[0006] This invention provides a method for detecting via events, the method comprising:

[0007] The ion current between the first cavity and the second cavity is measured to obtain a preset number of ion current values; wherein, the first cavity is filled with a base sequence;

[0008] The preset number of ion current values ​​are divided into multiple sets of current values;

[0009] Peak values ​​are extracted from the multiple sets of current values ​​to determine the multiple current peak values ​​corresponding to the multiple sets of current values;

[0010] Based on the multiple current peaks and the baseline current values ​​corresponding to each of the multiple current value sets, the current amplitude corresponding to each of the multiple current peaks is determined;

[0011] The detection threshold is determined based on the current amplitude corresponding to each of the multiple current peaks;

[0012] Based on the current amplitude corresponding to each of the plurality of current peaks and the detection threshold, at least one via event of the base sequence is determined.

[0013] This invention provides an apparatus for detecting via events, the apparatus comprising: a data acquisition unit, a data segmentation unit, and a data determination unit; wherein,

[0014] The acquisition unit is used to measure the ion current between the first cavity and the second cavity to obtain a preset number of ion current values; wherein, the first cavity is filled with a base sequence;

[0015] The dividing unit is used to divide the preset number of ion current values ​​into multiple current value sets;

[0016] The determining unit is used to extract the peak values ​​of the plurality of current value sets respectively, and to determine the plurality of current peak values ​​corresponding to the plurality of current value sets;

[0017] The determining unit is further configured to determine the current amplitude corresponding to each of the plurality of current peaks based on the plurality of current peaks and the baseline current values ​​corresponding to each of the plurality of current value sets;

[0018] The determining unit is further configured to determine a detection threshold based on the current amplitude corresponding to each of the plurality of current peaks;

[0019] The determining unit is further configured to determine at least one via event of the base sequence based on the current amplitude corresponding to each of the plurality of current peaks and the detection threshold.

[0020] This invention provides an electronic device, the electronic device comprising:

[0021] Memory, used to store executable instructions;

[0022] When a processor executes executable instructions stored in the memory, the processor executes the method for detecting via events described above.

[0023] This invention provides a storage medium storing executable instructions. When the executable instructions are executed by one or more processors, the processors execute the via event detection method.

[0024] This invention provides a method, apparatus, electronic device, and storage medium for detecting via events. The method includes: measuring the ion current between a first cavity and a second cavity to obtain a preset number of ion current values; wherein the first cavity is filled with a base sequence; dividing the preset number of ion current values ​​into multiple current value sets; extracting peak values ​​from each of the multiple current value sets to determine multiple current peak values ​​corresponding to the multiple current value sets; determining the current amplitude corresponding to each of the multiple current peak values ​​based on the multiple current peak values ​​and the baseline current values ​​corresponding to each of the multiple current value sets; determining a detection threshold based on the current amplitude corresponding to each of the multiple current peak values; and determining at least one via event of the base sequence based on the current amplitude corresponding to each of the multiple current peak values ​​and the detection threshold. This method, on the one hand, treats via events as abnormal information, eliminating the need for manually setting thresholds from the perspective of the data itself, and automatically detecting via events, thus improving the efficiency of via event detection. On the other hand, the ion current values ​​are divided into multiple sets, each with a corresponding baseline current value. The amplitude is calculated based on the baseline current value to determine whether a via event has occurred in each set, and then the total number of via events that occurred in the whole process is determined. This approach, which combines local and global processing, greatly reduces the amount of computation while improving the accuracy of detection. Attached Figure Description

[0025] Figure 1 A flowchart illustrating a method for detecting via events provided in an embodiment of the present invention. Figure 1 ;

[0026] Figure 2 A flowchart illustrating a method for detecting via events provided in an embodiment of the present invention. Figure 2 ;

[0027] Figure 3 A flowchart illustrating a method for detecting via events provided in an embodiment of the present invention. Figure 3 ;

[0028] Figure 4 A flowchart illustrating a method for detecting via events provided in an embodiment of the present invention. Figure 4 ;

[0029] Figure 5 This is a schematic diagram illustrating the detection signal comparison effect provided in an embodiment of the present invention;

[0030] Figure 6 A schematic diagram illustrating the effects of baseline calculation, rising phase, and falling phase examples provided in embodiments of the present invention;

[0031] Figure 7 This is a schematic diagram illustrating the performance of the invention on multiple raw sequencing data in an embodiment of the invention;

[0032] Figure 8 This is a schematic diagram illustrating the effect of the ion current IV curve of the dielectric perforated nanopore provided in an embodiment of the present invention.

[0033] Figure 9 This is a schematic diagram illustrating the principle of the silicon nitride solid-state nanopore sensor provided in an embodiment of the present invention.

[0034] Figure 10 This is a schematic diagram of a device for detecting via events provided in an embodiment of the present invention;

[0035] Figure 11 This is a schematic diagram of the structure of an electronic device provided in an embodiment of the present invention. Detailed Implementation

[0036] The technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the protection scope of the present invention.

[0037] To enable those skilled in the art to better understand the present invention, the invention will be further described in detail below with reference to the accompanying drawings and specific embodiments. The method for detecting via events provided in this invention is applied to electronic devices. Figure 1 A flowchart illustrating a method for detecting via events provided in an embodiment of the present invention. Figure 1 , will combine Figure 1 The steps shown are explained.

[0038] S101. The ion current between the first cavity and the second cavity is measured to obtain a preset number of ion current values; wherein, the first cavity is filled with a base sequence.

[0039] In an embodiment of the present invention, a 60°C water bath is used to heat the single-stranded DNA sample to facilitate uniform dispersion of the single-stranded DNA molecules. After cooling to room temperature, the single-stranded DNA molecules are added to the first chamber in the liquid pool. A positive voltage is applied to the first chamber and a negative voltage is applied to the second chamber to obtain ion current values ​​at different times. A preset number of ion current values ​​are obtained by measurement using a patch-clamp amplifier.

[0040] In an embodiment of the present invention, the first cavity can be a reverse cavity and the second cavity can be a normal cavity. The normal cavity and the reverse cavity are obtained by separating the liquid pool by the punched chip.

[0041] S102. Divide a preset number of ion current values ​​into multiple current value sets.

[0042] In some embodiments of the present invention, the electronic device divides the acquired ion current values ​​into multiple sets of current values, each set including current values ​​at different times.

[0043] In some embodiments of the present invention, a preset number of ion current values ​​can be randomly divided into multiple sets of current values. Experiments are then conducted to verify the results, and the number of sets of current values ​​with the best final performance is selected for further division. Alternatively, a time interval can be preset, and the preset ion current values ​​can be divided according to the time interval; the present invention does not impose any limitations on this method.

[0044] S103. Extract the peak values ​​from the multiple current value sets respectively, and determine the multiple current peak values ​​corresponding to the multiple current value sets.

[0045] In an embodiment of the present invention, the electronic device determines the largest current value in each set of current values ​​to obtain multiple current peak values ​​corresponding to multiple sets of current values.

[0046] S104. Based on the baseline current values ​​corresponding to multiple current peaks and multiple sets of current values, determine the current amplitude corresponding to each of the multiple current peaks.

[0047] In an embodiment of the present invention, after the electronic device acquires multiple current peaks, it calculates the baseline current value corresponding to each current value set, and calculates the current amplitude corresponding to each current set based on the baseline current value and the current peak.

[0048] In embodiments of the present invention, the baseline current value is the average current value over a preset time period. The current amplitude is the range of increase in the peak current value relative to the baseline current value, so the current amplitude can be obtained by the difference between the peak current value and the baseline current value.

[0049] S105. Determine the detection threshold based on the current amplitude corresponding to each of the multiple current peaks.

[0050] In an embodiment of the present invention, the electronic device calculates a detection threshold based on the acquired current amplitude.

[0051] S106. Based on the current amplitude and detection threshold corresponding to each of the multiple current peaks, determine at least one via event of the base sequence.

[0052] In an embodiment of the present invention, after the electronic device obtains a detection threshold, it compares the current amplitude corresponding to each of the multiple current peaks with the detection threshold to determine the amplitude that meets the condition. This determined amplitude indicates that DNA has passed through the nanopore, i.e., a perporeation event has occurred.

[0053] In embodiments of the present invention, a perforation event refers to a significant change in current caused by the passage of DNA sequences through nanopores. Specifically, when bases pass through solid pores, the current changes, thereby temporarily affecting the current intensity flowing through the nanopores. Compared with the current value when no DNA passes through, the current value will change to a certain extent within a certain range when DNA passes through.

[0054] It is understood that, in the embodiments of the present invention, a preset number of ion current values ​​are collected; the electronic device divides the collected preset number of ion current values ​​into multiple current value sets; the current peak value is extracted from each current value set; based on the current peak value and baseline current value of each current value set, the corresponding current amplitude is obtained; based on multiple current amplitudes, a detection threshold is obtained; and based on the current amplitude and detection threshold, at least one via event of the base sequence is determined. This method, on the one hand, treats via events as abnormal information, automatically detecting via events without requiring manual threshold setting, thus improving the efficiency of via event detection. On the other hand, dividing the ion current values ​​into multiple sets, each with a corresponding baseline current value, calculating the amplitude based on the baseline current value, determining whether a via event has occurred in each set, and then determining the total number of via events occurring throughout the process—this combined local and global processing approach greatly reduces computational load while improving detection accuracy.

[0055] S103 can be implemented through S1031 to S1033. Figure 2 A flowchart illustrating a method for detecting via events provided in an embodiment of the present invention. Figure 2 , will combine Figure 2 The steps shown are explained.

[0056] S1031. Sort the current values ​​in the multiple current value sets to obtain multiple sorting results corresponding to the multiple current sets.

[0057] In some embodiments of the present invention, the electronic device sorts the current values ​​in each set of current values ​​and determines the sorting result in each set of current values.

[0058] In some embodiments of the present invention, existing techniques may be used for sorting, and the present invention does not impose any limitations.

[0059] S1032. Among multiple sorting results, obtain multiple maximum current values ​​corresponding to multiple current sets.

[0060] In some embodiments of the present invention, the electronic device extracts the maximum value from each sorting result. For multiple sorting results, multiple maximum current values ​​will ultimately be obtained.

[0061] S1033. Determine multiple maximum current values ​​as multiple current peak values ​​corresponding to multiple current value sets.

[0062] In some embodiments of the present invention, after the electronic device obtains the maximum value of each current value set, it uses the maximum value as the peak value of each current value set.

[0063] It is understood that, in some embodiments of the present invention, the current values ​​in multiple sets of current values ​​are sorted separately, and the maximum value is extracted from each of the multiple sets of current values ​​in the sorting results. This method improves the accuracy and efficiency of extracting the maximum value from the set of current values.

[0064] In some embodiments of the present invention, before S104, the method for detecting via events further includes S201, which can be implemented by S2011 to S2012. Figure 3 A flowchart illustrating a method for detecting via events provided in an embodiment of the present invention. Figure 3 , will combine Figure 3 The steps shown are explained.

[0065] S2011. Obtain the time points corresponding to each of the multiple current peak values.

[0066] In some embodiments of the present invention, the electronic device acquires the time point corresponding to each current peak.

[0067] S2012. Based on the time point, determine the baseline current value corresponding to each of the multiple current value sets from the ion current values ​​contained in the multiple current value sets.

[0068] In some embodiments of the present invention, the electronic device obtains current values ​​within a preset range based on the time point of the current peak in each current value set, and calculates the baseline current value of each current value set based on the current values ​​within the range.

[0069] It is understood that, in some embodiments of the present invention, multiple current peak values ​​are obtained at their respective time points, and based on these time points, baseline current values ​​corresponding to multiple sets of current values ​​are determined. This method calculates the baseline current value for each segment by calculating the points near the peak values ​​in each segment, thus solving the problem that accurate event detection cannot be performed using the mean and variance of a large number of data points as thresholds due to variations in baseline current.

[0070] In some embodiments of the present invention, S104 can be implemented by S1041, which will be described in detail in conjunction with the following steps.

[0071] S1041. The difference between the baseline current value and the peak current value corresponding to each of the multiple current value sets is determined as the current amplitude corresponding to each of the multiple peak current values.

[0072] In some embodiments of the present invention, the electronic device obtains the current amplitude corresponding to each current peak by subtracting the corresponding baseline current value from the current peak value corresponding to each set in a plurality of current value sets.

[0073] In some embodiments of the present invention, if the peak current is represented by p, the baseline current value is represented by b, and the current amplitude is represented by a, then the current amplitude corresponding to each peak current is calculated as a = pb.

[0074] In some embodiments of the present invention, S2012, which has not been mentioned before, can be implemented through S301 to S304. Figure 4 A flowchart illustrating a method for detecting via events provided in an embodiment of the present invention. Figure 4 , will combine Figure 4 The steps shown are explained.

[0075] S301. Based on the time point, the first time threshold, and the second time threshold, determine the stable current time range corresponding to each of the multiple current value sets.

[0076] In some embodiments of the present invention, the electronic device determines a time range in each set of current values ​​based on the time point of the current peak value in each set of current values, as well as a preset first time threshold and a second time threshold. Within this time range, the current change is in a relatively stable state.

[0077] In some embodiments of the present invention, the first time threshold can be set to 1.5, the second time threshold can be set to 0.75, and vice versa.

[0078] In some embodiments of the present invention, within a fragment, the baseline current value, compared to the current peak of that fragment, is defined as the average current value over a steady-state period before the bases pass through the solid-state pore. According to literature reports, the average time for 22 nucleotides to pass through is approximately 0.2 ms, falling within the range of 1.5 ms to 0.75 ms before the peak, i.e., [k...]. i -1.5,k i [-0.75], the current change is in a relatively stable state, and the baseline current value can be calculated based on the current value in this range, where k is the time point of the current peak.

[0079] S302. Based on the stable current time range, extract multiple candidate current values ​​corresponding to multiple current value sets.

[0080] In some embodiments of the present invention, after the electronic device obtains a stable current time range in each set of current values, it extracts multiple current values ​​contained within that time range and uses these multiple current values ​​as candidate current values.

[0081] S303. Calculate the mean of multiple candidate current values ​​to obtain the mean of each set of multiple current values.

[0082] In some embodiments of the present invention, the electronic device calculates the mean of the candidate current values ​​for each set of current values ​​based on the corresponding candidate current values.

[0083] S304. The mean value corresponding to each of the multiple current value sets is determined as the baseline current value corresponding to each of the multiple current value sets.

[0084] In some embodiments of the present invention, after calculating the average value, the electronic device uses the average value as the baseline current value corresponding to each set of current values.

[0085] It is understood that, in some embodiments of the present invention, based on the time point of the current peak and a set threshold, a time range of stable current corresponding to each of multiple current value sets is determined. The average current value within this time range is calculated using the current values ​​within that range, and this average current value is used as the baseline current value corresponding to each of the multiple current value sets. This method differs from other methods in determining the baseline current value by using the average value over a stable period before the via event occurs, resulting in a more reasonable amplitude.

[0086] In some embodiments of the present invention, S105 can be implemented by S1051 to S1055, which will be described in detail in conjunction with the following steps.

[0087] S1051. Sort the current amplitudes corresponding to the multiple current peaks to obtain the sorting results.

[0088] In some embodiments of the present invention, the electronic device sorts the current amplitudes corresponding to multiple current values ​​to obtain a sorting result.

[0089] S1052. Determine the first quantile and the third quantile from the sorting results.

[0090] In some embodiments of the present invention, the first quantile and the third quantile are determined from the sorting results.

[0091] In some embodiments of the present invention, the first and third quartiles are related to the interquartile range (IQR). A quartile is a type of quantile in statistics, where all values ​​are arranged in ascending order and divided into four equal parts; the values ​​at the three dividing points are the quartiles. The difference between the third quartile and the first quartile is also called the interquartile range (IQR). The first quartile (Q1), also known as the "smaller quartile," is equal to the 25th percentile of all values ​​in the sample arranged in ascending order. The second quartile (Q2), also known as the "median," is equal to the 50th percentile of all values ​​in the sample arranged in ascending order; the third quartile (Q3), also known as the "larger quartile," is equal to the 75th percentile of all values ​​in the sample arranged in ascending order; the difference between the third quartile and the first quartile is also called the interquartile range (IQR).

[0092] S1053. Subtract the first quantile from the third quantile to obtain the difference result.

[0093] In some embodiments of the present invention, after the electronic device obtains the first quantile and the third quantile, it subtracts the first quantile from the third quantile to obtain the difference result.

[0094] In some embodiments of the present invention, if the first quantile is Q1 and the third quantile is Q3, and the difference is IQR = Q3 - Q1.

[0095] S1054. Magnify the difference result to obtain the magnified difference result.

[0096] In some embodiments of the present invention, the electronic device magnifies the acquired difference result by a factor of n to obtain the magnified difference result.

[0097] In some embodiments of the present invention, n can be 5.

[0098] S1055. Sum the expanded difference result with the third quantile to obtain the detection threshold.

[0099] In some embodiments of the present invention, the electronic device sums the amplified difference result with the third quantile to obtain the detection threshold.

[0100] In some embodiments of the present invention, in conjunction with S1052-S1055, if the detection threshold is set to y, then y can be obtained using y = Q3 + 5 * IQR.

[0101] It is understood that in some embodiments of the present invention, multiple amplitude values ​​are sorted, and a first quantile and a third quantile are determined from the sorting results. Based on the first quantile and the third quantile, a detection threshold is obtained, which can be used for subsequent detection of via events.

[0102] In some embodiments of the present invention, S106 can be obtained from S1061, as will be explained in detail in conjunction with the following steps.

[0103] S1061. When the current amplitude corresponding to each of the multiple current peaks is greater than the detection threshold, it is determined that a via event has occurred in the base sequence.

[0104] In some embodiments of the present invention, the electronic device compares the current amplitude corresponding to each current peak with a detection threshold, selects amplitudes greater than the detection threshold, and determines that the occurrence of these amplitudes greater than the detection threshold is due to DNA passing through the nanopore, thus confirming that a perforation event has occurred in the base sequence.

[0105] In some embodiments of the present invention, a is selected to satisfy a i The amplitude of >y = Q3 + 5 * IQR is obviously an outlier among all amplitudes. This invention believes that the occurrence of these outliers is due to DNA passing through the nanopore, and thus it is determined as an event point, which represents the occurrence of a perforation event.

[0106] For example, as shown in Table 1, Table 1 details multiple via events. Using the method of this invention, 51 via events were successfully identified from the current values ​​stored in a single ABF file. Table 1 includes the event index, peak time, and amplitude of each of the 51 via events.

[0107] (Amplitude), baseline current value, start time, end time, and total via duration. Using the method proposed in this invention, via event points can be located among multiple data points in the acquired data.

[0108] Table 1

[0109]

[0110] Figure 5 This is a schematic diagram illustrating the signal comparison effect. Figure 5In the diagram, the gray lines represent the trend of current change over time in the original data, the 51 marked vertical lines represent the event points automatically selected by the detection method of this invention in the entire signal, the light-colored circles represent events that have been manually confirmed and correctly identified by the detection method of this invention, and the black circles represent cases where the detection method of this invention selects incorrectly. Figure 6 This is a schematic diagram illustrating the effects of baseline calculation, rising phase, and falling phase examples. Figure 6 In the table, the black line segment is used to calculate the baseline current value, and the lightest, most prominent tip represents the time range of the via event and the current variation. Table 1 shows the performance on a single raw data point. Figure 7 This is a schematic diagram illustrating the performance across multiple raw sequencing datasets. Figure 7 In order to comprehensively evaluate the performance of the detection method of this invention, event detection was performed on all 23 ABF files. Experts were asked to annotate all event points, and two evaluation metrics, coverage and accuracy, were used: For each original signal file, coverage can be defined by formula (1), as shown below:

[0111]

[0112] Wherein, H is a set of events given manual annotation; A is a set of events detected by this invention.

[0113] The precision is expressed by formula (2), as shown below:

[0114]

[0115] In some embodiments of the present invention, coverage reflects whether the detection method of the present invention can accurately detect all real events, while precision reflects whether the events detected by the detection method of the present invention are real. Ideally, if the events detected by the detection method of the present invention are completely consistent with the events annotated by experts, then both the coverage and precision of that sample will be equal to 1. Figure 7 In the diagram, the upper part represents coverage, and the lower part represents accuracy. It can be seen that the detection method of this invention achieves an average coverage of 0.956, with a coverage of 1 in 9 of the ABF files, indicating its excellent performance in detecting real events. Simultaneously, its average coverage also reaches 0.876. Some events identified by the detection method of this invention were not accepted by experts. This is mainly because, although individual amplitudes were indeed outliers among all amplitudes, these amplitudes were slightly below the expert's threshold. Since statistical analysis of 10,000 amplitudes is required in each ABF file, the fact that some amplitude outliers were not accepted by experts is acceptable.

[0116] Understandably, in some embodiments of the present invention, the current amplitudes corresponding to multiple current peaks are compared, and the amplitude greater than a detection threshold is selected. This amplitude indicates that a base sequence via event has occurred in the corresponding set of current values. This method compares the current amplitude corresponding to each current value with the detection threshold, and then determines the via event based on the amplitudes corresponding to all peaks. It adopts a processing idea that combines local and global features, which greatly reduces the amount of computation and yields more reliable results.

[0117] In some embodiments of the present invention, S1061 is followed by S401-S405, which will be described in detail in conjunction with the following steps.

[0118] S401. From multiple sets of current values, obtain at least one set of matching current values ​​corresponding to each via event.

[0119] In some embodiments of the present invention, the electronic device obtains a matching current value set corresponding to each via event in each current value set. The set includes the current value set corresponding to the via event, as well as the current peak value, baseline current value and amplitude in the current value set.

[0120] S402. Based on the matching time point of the peak value of the matching current in the matching current value set, divide the time points corresponding to each matching current value in the matching current value set into a first time point set greater than the matching time point and a second time point set less than the matching time point.

[0121] In some embodiments of the present invention, the electronic device divides the set of matching current values ​​into two sets based on the time point of the configured current peak: a set of current values ​​greater than the time point and a set of current values ​​less than the time point.

[0122] In some embodiments of the present invention, for a via event, since it corresponds to a local maximum current value, there must be a rise period before the peak and a decay period after the peak. The sum of these two times represents the total time from the start of base entry into the solid-state via to its final exit. Furthermore, since the baseline value is calculated first using the mean of the stable phase before the rise period, there must be data points with values ​​higher than the peak value before and after the peak. Therefore, the set of current values ​​corresponding to the via event is first divided into two regions using the time point of the current peak, and the current values ​​in these two regions are compared with the baseline current value.

[0123] S403. In the second time point set, determine the start time corresponding to at least one via event; wherein the matching current value corresponding to the start time is the Nth value that is greater than the matching baseline current in the matching current value set; N is a positive integer.

[0124] In some embodiments of the present invention, the electronic device finds the Nth current value that is greater than the baseline current value at a time point less than the peak value of the matching current, and uses the time point of that current value as the start time of the via event.

[0125] S404. In the first time point set, determine the end time corresponding to at least one via event; wherein, the current value corresponding to the end time is the Mth value of the expanded current value corresponding to the matching baseline current that is greater than the peak value of the matching current; M is a positive integer.

[0126] In some embodiments of the present invention, the electronic device searches for the scaled current value corresponding to the Mth baseline current value after the peak of the matching current in a set of time points greater than the peak of the matching current.

[0127] In some embodiments of the present invention, the scaling current value can be 0.1*ai, and the scaling current value corresponding to the baseline current value is bi+0.1*ai.

[0128] S405. Determine the via time corresponding to each of the at least one via event based on the start time and end time of each via event.

[0129] In some embodiments of the present invention, the electronic device obtains the via time corresponding to each via event by subtracting the end time from the start time corresponding to each via event.

[0130] In some embodiments of the present invention, S501 to S503 are included before S101, which will be described in detail in conjunction with the following steps.

[0131] S501. Apply a pulse voltage to the chip placed in the preset electrolyte solution through a numerical source meter, and monitor the system current to obtain the monitored current value.

[0132] In some embodiments of the present invention, the cleaned, un-drilled chip is clamped between two polymethyl methacrylate (PMMA) gaskets using silicone rubber gaskets. Both ends of the liquid pool are filled with 1M KCl electrolyte solution. Then, a pulse voltage is applied to the chip using a numerical source meter (Keithley 2612A), and the system current value is monitored to obtain the monitored current value.

[0133] In some embodiments of the present invention, the un-drilled silicon nitride chip is cleaned using a standardized cleaning procedure before being placed in an electrolyte solution. First, the un-drilled silicon nitride chip is placed in a piranha solution (concentrated sulfuric acid / hydrogen peroxide = 3:1), heated at 180°C for 30 minutes, cooled to room temperature, and then repeatedly cleaned with anhydrous ethanol and deionized water.

[0134] S502. When the monitored current value is greater than the current threshold, stop applying the pulse voltage to obtain the punched chip; the punched chip is used to divide the liquid pool into a first cavity and a second cavity; the first cavity and the second cavity are two independent liquid storage chambers.

[0135] In some embodiments of the present invention, the monitored current value is compared with a preset current threshold. If the monitored current value is greater than the current threshold, the applied pulse voltage is stopped, resulting in a punched chip. This punched chip can divide the liquid pool into two independent liquid storage chambers: a first chamber and a second chamber.

[0136] In some embodiments of the present invention, the first cavity can be a reverse cavity and the second cavity can be a cis cavity.

[0137] In some embodiments of the present invention, a 20Hz, 13-20V pulse voltage can be applied to both ends of the silicon nitride chip using a numerical source meter (Keithley 2612A). The pulse voltage is stopped when the current detected by the numerical source meter increases significantly, thus completing the dielectric perforation fabrication of silicon nitride nanopores. After dielectric perforation is completed, the ion current (IV) curve of the silicon nitride nanopores is measured using a 1M KCl solution, and the pore size of the nanopores is calculated, as shown in Figure 8. Figure 8 This is a schematic diagram of the ion current (IV) curve of a dielectric perforated nanopore. The horizontal axis represents voltage, the vertical axis represents current, and 5.56 nm represents the pore size.

[0138] In some embodiments of the present invention, one of the key aspects of solid-state nanopore sequencing technology is the design of a special nanoscale pore. When DNA bases pass through the nanopore, they cause a change in charge, thereby briefly affecting the current intensity flowing through the nanopore. Sensitive electronic devices capture these changes to determine the bases that have passed through. Figure 9 As shown, Figure 9 This is a schematic diagram illustrating the principle of a silicon nitride solid-state nanopore sensor. The chip is placed in a KCl electrolyte solution, and the device detects the resulting charge change as the base passes through the nanopore. Figure 9 It includes electricity meters, as well as silicon, silicon dioxide, chloride ions, potassium ions, and silicon nitride.

[0139] S503. A positive voltage is applied to the first cavity filled with the base sequence, and a negative voltage is applied to the second cavity to obtain an ion current.

[0140] In some embodiments of the present invention, a base sequence is added to a first cavity, a positive voltage is applied to the first cavity, and a negative voltage is applied to a second cavity, causing the anions and cations in the solution to move in a specific direction, thereby generating an ion current value.

[0141] In some embodiments of the present invention, S101 can be implemented by S1011, which will be described in detail in conjunction with the following steps.

[0142] S1011. Use a patch clamp amplifier to measure the ion current at a preset sampling rate until a specified time is reached, and obtain a preset number of ion current values.

[0143] In some embodiments of the present invention, after the ion current is generated, the ion current is measured using a patch clamp amplifier at a preset sampling rate to obtain a preset number of ion current values.

[0144] In some embodiments of the present invention, the patch clamp amplifier can perform measurements at a sampling rate of 250 kHz with a low-pass filter applied at 10 kHz.

[0145] In some embodiments of the present invention, S102 can be implemented by S601-S603, which will be described in detail in conjunction with the following steps.

[0146] S601. Write a preset number of ion current values ​​into a file with a preset format to obtain a current data file.

[0147] In some embodiments of the present invention, after a preset ion current value is collected, the current value is stored in a file in a preset format to obtain a current data file.

[0148] In some embodiments of the present invention, the preset format file can be an abf file.

[0149] S602. The current data file is split multiple times to obtain multiple file segments.

[0150] In some embodiments of the present invention, the electronic device splits the file into multiple segments on an average basis.

[0151] In some embodiments of the present invention, a file may be split into 10,000 segments.

[0152] In some embodiments of the present invention, the file partitioning strategy is the first step and a crucial factor in the event detection method, as it directly affects the performance and computation time of the proposed detection method. Too many partitions increase computation time and may result in the same event's data points being divided into multiple partitions, leading to false positives. Conversely, fewer partitions, while reducing computation time, may result in multiple events being grouped into the same partition, leading to missed detections.

[0153] For example, Table 2 shows the impact of various partitioning strategies on the results analyzed by this invention.

[0154] Table 2

[0155] 100 300 0.168 0.873 5.691 500 600 0.591 0.95 5.804 1000 300 0.728 0.951 5.988 5000 60 0.943 0.918 7 8000 37.5 0.958 0.89 7.745 10000 30 0.962 0.885 8.25 12000 25 0.962 0.885 8.741 15000 20 0.964 0.885 9.501 20000 15 0.969 0.876 10.734 30000 10 0.962 0.901 13.275 50000 6 0.95 0.923 18.409 100000 3 0.864 0.916 30.683 500000 0.6 0.786 0.88 94.873

[0156] As shown in Table 2, the number of segments has little impact on the final result when the number of segments is between 5000 and 50000. However, when the number of segments is between 100 and 1000, the duration of each segment is too long, potentially containing multiple via events. Since we only focus on the peak value within a segment, at most one event can be detected per segment, leading to performance degradation. Because via events are relatively sparse, when there are too many segments, such as 100,000 to 500,000, the number of potential event points also increases, but the proportion of actual via events is extremely low. These low-amplitude noise signals reduce the calculated quartile values, increasing the number of incorrectly identified events and decreasing both coverage and accuracy. Furthermore, the computation time required increases significantly with the number of segments. To balance performance and computational efficiency, this invention ultimately chooses 30ms as one segment.

[0157] S603. Use the ion current values ​​contained in each of the multiple file fragments to form multiple sets of current values.

[0158] In some embodiments of the present invention, each file fragment contains multiple ion current values, and one file fragment is equivalent to a set of current values.

[0159] In some embodiments of the present invention, the data file can be split into 10,000 segments, each segment including 7,500 data points (time interval 30ms). Each data point is a two-dimensional array containing the time point and the current value collected at that time point.

[0160] It is understood that in some embodiments of the present invention, a preset number of ion current values ​​are collected; the collected preset number of ion current values ​​are divided into multiple current value sets; the current peak value is extracted from each current value set; the corresponding current amplitude is obtained based on the current peak value and the baseline current value of each current value set; a detection threshold is obtained based on multiple current amplitudes; and at least one via event of the base sequence is determined based on the current amplitude and the detection threshold. This method, on the one hand, treats via events as abnormal information, automatically detecting via events without requiring manual threshold setting from the perspective of the data itself, thus improving the efficiency of via event detection. On the other hand, dividing the ion current values ​​into multiple sets, each with a corresponding baseline current value, calculating the amplitude based on the baseline current value, determining whether a via event has occurred in each set, and then determining the total number of via events occurring throughout the process—this combined local and global processing approach greatly reduces computational load while improving detection accuracy.

[0161] This invention provides a method for detecting via events, which will be described in detail below.

[0162] S1. Collect the preset ion current value.

[0163] In an embodiment of the present invention, S1 can also be implemented in the following manner: cleaning the un-drilled silicon chloride chip; placing the cleaned silicon chloride chip in an electrolyte solution; applying a voltage to the silicon chloride chip using a numerical source meter, stopping the application of the pulse voltage when the numerical source meter detects an increase in the system current, thus obtaining a drilled silicon chloride chip; the drilled silicon chloride chip divides the liquid pool into two independent storage chambers, resulting in a trans chamber and a cis chamber; placing DNA into the trans chamber, applying a positive voltage to the trans chamber and a negative voltage to the cis chamber, thus obtaining a current trajectory; measuring the current trajectory using a patch-clamp amplifier to obtain ion current values ​​at different times within a specified time period;

[0164] S2. Save the collected ion current values ​​to an abf file and divide the file into multiple segments, each segment containing ion current values ​​at different time points.

[0165] S3. Extract the peak current value corresponding to each segment from the current values ​​of each segment.

[0166] S4. Calculate the time range corresponding to each of the multiple segments based on the current peak value, the first time threshold, and the second time threshold.

[0167] S5. Calculate the baseline current value corresponding to each segment by using the current values ​​contained within a time range corresponding to each segment.

[0168] S6. Subtract the peak current value from the baseline current value to obtain the amplitude corresponding to each of the multiple segments.

[0169] S7. Compare the amplitude and detection threshold corresponding to each of the multiple fragments to determine at least one via event of the base sequence.

[0170] S8. After determining at least one via event, obtain the current peak value corresponding to the via event, determine the first current value greater than the baseline current value before the current peak value, obtain the time point corresponding to the current value greater than the baseline current value, and determine it as the start time.

[0171] S9. Based on the current peak value corresponding to the via event, determine the end time after the current peak value according to preset conditions.

[0172] S10. Calculate the via time corresponding to the via event based on the start time and end time.

[0173] Understandably, in the embodiments of the present invention, after the ion current values ​​are collected, they are divided into multiple current value sets; the current peak value is extracted from each current value set; based on the current peak value and the baseline current value of each current value set, the corresponding current amplitude is obtained; based on multiple current amplitudes, a detection threshold is obtained; and based on the current amplitude and the detection threshold, at least one via event of the base sequence is determined. This method, on the one hand, treats via events as abnormal information, automatically detecting via events without requiring manual threshold setting, thus improving the efficiency of via event detection. On the other hand, dividing the ion current values ​​into multiple sets, each with a corresponding baseline current value, and calculating the amplitude based on the baseline current value, determines whether a via event has occurred in each set, thereby determining the total number of via events occurring throughout the process. This combined local and global processing approach greatly reduces computational load while improving detection accuracy.

[0174] This invention provides a device for detecting via events, such as... Figure 10 As shown, Figure 10 This is a schematic diagram of a via event detection device provided in an embodiment of the present invention. The via event detection device 10 includes: a data acquisition unit 1001, a segmentation unit 1002, and a determination unit 1003; wherein,

[0175] The acquisition unit 1001 is used to measure the ion current between the first cavity and the second cavity to obtain a preset number of ion current values; wherein, the first cavity is filled with a base sequence.

[0176] The division unit 1002 is used to divide the preset number of ion current values ​​into multiple current value sets.

[0177] The determining unit 1003 is used to extract the peak values ​​of the multiple current value sets respectively, and determine the multiple current peak values ​​corresponding to the multiple current value sets.

[0178] The determining unit 1003 is further configured to determine the current amplitude corresponding to each of the plurality of current peaks based on the plurality of current peaks and the baseline current values ​​corresponding to each of the plurality of current value sets.

[0179] The determining unit 1003 is further configured to determine a detection threshold based on the current amplitude corresponding to each of the plurality of current peaks.

[0180] The determining unit 1003 is further configured to determine at least one via event of the base sequence based on the current amplitude corresponding to each of the plurality of current peaks and the detection threshold.

[0181] In some embodiments of the present invention, the determining unit 1003 is further configured to sort the current values ​​in the plurality of current value sets to obtain a plurality of sorting results corresponding to the plurality of current sets; obtain a plurality of maximum current values ​​corresponding to the plurality of current sets from the plurality of sorting results; and determine the plurality of maximum current values ​​as a plurality of current peak values ​​corresponding to the plurality of current value sets.

[0182] In some embodiments of the present invention, the determining unit 1003 is further configured to obtain the time point corresponding to each of the plurality of current peaks before determining the current amplitude corresponding to each of the plurality of current peaks based on the corresponding baseline current value of each of the plurality of current value sets; and based on the time point, determine the baseline current value corresponding to each of the plurality of current value sets from the ion current values ​​included in the plurality of current value sets.

[0183] In some embodiments of the present invention, the determining unit 1003 is further configured to determine the difference between the baseline current value corresponding to each of the plurality of current value sets and the plurality of current peak values ​​as the current amplitude corresponding to each of the plurality of current peak values.

[0184] In some embodiments of the present invention, the determining unit 1003 is further configured to: determine the stable current time range corresponding to each of the plurality of current value sets according to the time point, the first time threshold, and the second time threshold; extract a plurality of candidate current values ​​corresponding to each of the plurality of current value sets according to the stable current time range; calculate the mean of the plurality of candidate current values ​​to obtain the mean of each of the plurality of current value sets; and determine the mean of each of the plurality of current value sets as the baseline current value corresponding to each of the plurality of current value sets.

[0185] In some embodiments of the present invention, the determining unit 1003 is further configured to sort the current amplitudes corresponding to the plurality of current peaks to obtain a sorting result; determine a first quantile and a third quantile from the sorting result; subtract the first quantile and the third quantile to obtain a difference result; expand the difference result to obtain an expanded difference result; and sum the expanded difference result and the third quantile to obtain the detection threshold.

[0186] In some embodiments of the present invention, the determining unit 1003 is further configured to determine that a via event has occurred in the base sequence when the current amplitude corresponding to each of the plurality of current peaks is greater than the detection threshold.

[0187] In some embodiments of the present invention, the determining unit 1003 is further configured to: divide the time points corresponding to each matching current value in the matching current value set into a first time point set greater than the matching time point and a second time point set less than the matching time point, based on the matching time point of the peak matching current in the matching current value set; and determine the start time corresponding to each of the at least one via event in the second time point set, wherein the matching current value corresponding to the start time is greater than the Nth value of the matching baseline current in the matching current value set; N is a positive integer; and determine the end time corresponding to each of the at least one via event in the first time point set, wherein the current value corresponding to the end time is greater than the Mth value of the expanded current value corresponding to the matching baseline current; M is a positive integer; and determine the via time corresponding to each of the at least one via event based on the start time and the end time corresponding to each of the at least one via event.

[0188] In some embodiments of the present invention, before measuring the ion current between the first cavity and the second cavity to obtain a preset number of ion current values, the acquisition unit 1001 is further configured to apply a pulse voltage to a chip placed in a preset electrolyte solution through a numerical source table and monitor the system current to obtain a monitoring current value; when the monitoring current value is greater than a current threshold, the application of the pulse voltage is stopped, and a perforated chip is obtained; the perforated chip is used to divide the liquid pool into the first cavity and the second cavity; the first cavity and the second cavity are two independent liquid storage chambers; a positive voltage is applied to the first cavity filled with a base sequence, and a negative voltage is applied to the second cavity to obtain the ion current.

[0189] In some embodiments of the present invention, the acquisition unit 1001 is further configured to use a patch clamp amplifier to measure the ion current at a preset sampling rate until a predetermined time is reached, thereby obtaining the preset number of ion current values.

[0190] In some embodiments of the present invention, the partitioning unit 1002 is further configured to write the preset number of ion current values ​​into a file of preset format to obtain a current data file; to split the current data file multiple times to obtain multiple file segments; and to use the ion current values ​​contained in each of the multiple file segments to form the multiple current value sets.

[0191] Understandably, in the above-described device implementation, after collecting the ion current values, they are divided into multiple current value sets; the peak current value is extracted from each current value set; based on the peak current value and the baseline current value of each current value set, the corresponding current amplitude is obtained; based on multiple current amplitudes, a detection threshold is obtained; and based on the current amplitude and the detection threshold, at least one via event of the base sequence is determined. This method, on the one hand, treats via events as abnormal information, automatically detecting via events without requiring manual threshold setting, thus improving the efficiency of via event detection. On the other hand, dividing the ion current values ​​into multiple sets, each with a corresponding baseline current value, and calculating the amplitude based on the baseline current value, determines whether a via event has occurred in each set, thereby determining the total number of via events occurring throughout the process. This combined local and global processing approach greatly reduces computational load while improving detection accuracy.

[0192] Based on the methods described in the above embodiments, an embodiment of the present invention provides a structural schematic diagram, as follows: Figure 11 As shown, Figure 11 The present invention provides a schematic diagram of a via event detection device. The via event detection device 11 includes a processor 1101 and a memory 1102. The memory 1102 stores one or more programs executable by the processor 1101. When one or more programs are executed, the processor 1101 executes a via event detection method as described in the previous embodiment.

[0193] This invention provides a computer-readable storage medium storing executable instructions for inducing a processor to execute the method for detecting via events.

[0194] Those skilled in the art will understand that embodiments of the present invention can be provided as methods, systems, or computer program products. Therefore, the present invention can take the form of hardware embodiments, software embodiments, or embodiments combining software and hardware aspects. Furthermore, the present invention can take the form of a computer program product embodied on one or more computer-usable storage media (including, but not limited to, disk storage and optical storage) containing computer-usable program code.

[0195] This invention is described with reference to flowchart illustrations and / or block diagrams of methods, apparatus (systems), and computer program products according to embodiments of the invention. It will be understood that each block of the flowchart illustrations and / or block diagrams, and combinations of blocks in the flowchart illustrations and / or block diagrams, can be implemented by computer program instructions. These computer program instructions can be provided to a processor of a general-purpose computer, special-purpose computer, embedded processor, or other programmable data processing apparatus to produce a machine, such that the instructions, which execute via the processor of the computer or other programmable data processing apparatus, generate instructions for implementing the flowchart illustrations and / or block diagrams. Figure 1 One or more processes and / or boxes Figure 1 A device that provides the functions specified in one or more boxes.

[0196] These computer program instructions may also be stored in a computer-readable storage medium that can direct a computer or other programmable data processing device to function in a particular manner, such that the instructions stored in the computer-readable storage medium produce an article of manufacture including instruction means, which are implemented in a process Figure 1 One or more processes and / or boxes Figure 1 The function specified in one or more boxes.

[0197] These computer program instructions may also be loaded onto a computer or other programmable data processing equipment to cause a series of operational steps to be performed on the computer or other programmable equipment to produce a computer-implemented process, thereby providing instructions that execute on the computer or other programmable equipment for implementing the process. Figure 1 One or more processes and / or boxes Figure 1 The steps of the function specified in one or more boxes.

[0198] The above description is merely a preferred embodiment of the present invention and is not intended to limit the scope of protection of the present invention.

Claims

1. A method for detecting via events, characterized in that, include: The ion current between the first cavity and the second cavity is measured to obtain a preset number of ion current values; wherein, the first cavity is filled with a base sequence; The preset number of ion current values ​​are divided into multiple sets of current values; Peak values ​​are extracted from the multiple sets of current values ​​to determine the multiple current peak values ​​corresponding to the multiple sets of current values; Obtain the time points corresponding to each of the multiple current peak values; Based on the time point, the first time threshold, and the second time threshold, determine the stable current time range corresponding to each of the multiple current value sets; Based on the stable current time range, extract multiple candidate current values ​​corresponding to the multiple current value sets; The mean of the multiple candidate current values ​​is calculated to obtain the mean of each set of multiple current values. The mean value corresponding to each of the multiple sets of current values ​​is determined as the baseline current value corresponding to each of the multiple sets of current values; Based on the multiple current peaks and the baseline current values ​​corresponding to each of the multiple current value sets, the current amplitude corresponding to each of the multiple current peaks is determined; The current amplitudes corresponding to each of the multiple current peaks are sorted to obtain the sorting result; From the sorting results, determine the first quantile and the third quantile; The difference between the first quantile and the third quantile is obtained; The difference result is then amplified to obtain the amplified difference result; The detection threshold is obtained by summing the expanded difference result with the third quantile. When the current amplitude corresponding to each of the multiple current peaks is greater than the detection threshold, it is determined that a via event has occurred in the base sequence.

2. The method according to claim 1, characterized in that, The step of extracting peak values ​​from the multiple sets of current values ​​to determine multiple current peak values ​​corresponding to the multiple sets of current values ​​includes: The current values ​​in the multiple sets of current values ​​are sorted to obtain multiple sorting results corresponding to the multiple sets of current values; From the multiple sorting results, obtain the multiple maximum current values ​​corresponding to the multiple sets of current values; The plurality of maximum current values ​​are determined as the plurality of current peak values ​​corresponding to the plurality of current value sets.

3. The method according to claim 1, characterized in that, The step of determining the current amplitude corresponding to each of the plurality of current peaks based on the plurality of current peaks and the baseline current values ​​corresponding to each of the plurality of current value sets includes: The difference between the baseline current value corresponding to each of the plurality of current value sets and the plurality of current peak values ​​is determined as the current amplitude corresponding to each of the plurality of current peak values.

4. The method according to any one of claims 1-3, characterized in that, After determining at least one via event of the base sequence based on the current amplitude corresponding to each of the plurality of current peaks and the detection threshold, the method further includes: From the plurality of current value sets, obtain the matching current value set corresponding to each of the at least one via event; Based on the matching time point of the peak value of the matching current in the set of matching current values, the time points corresponding to each matching current value in the set of matching current values ​​are divided into a first set of time points greater than the matching time point and a second set of time points less than the matching time point. In the second set of time points, the start time corresponding to each of the at least one via event is determined; wherein, the matching current value corresponding to the start time is the Nth value that is greater than the matching baseline current in the set of matching current values; N is a positive integer; In the first set of time points, the end time corresponding to each of the at least one via event is determined; wherein, the current value corresponding to the end time is the Mth value that is greater than the amplified current value corresponding to the matching baseline current; M is a positive integer; The via time corresponding to each of the at least one via event is determined based on the start time and the end time corresponding to each of the at least one via event.

5. The method according to claim 1, characterized in that, Before measuring the ion current between the first cavity and the second cavity to obtain a preset number of ion current values, the method further includes: A pulse voltage is applied to the chip placed in a preset electrolyte solution using a numerical source meter, and the system current is monitored to obtain the monitored current value. When the monitored current value is greater than the current threshold, the pulse voltage is stopped, and the punched chip is obtained; the punched chip is used to divide the liquid pool into the first cavity and the second cavity; the first cavity and the second cavity are two independent liquid storage chambers; A positive voltage is applied to the first cavity filled with a base sequence, and a negative voltage is applied to the second cavity to obtain an ion current.

6. The method according to claim 1, characterized in that, The measurement of the ion current between the first cavity and the second cavity to obtain a preset number of ion current values ​​includes: The ion current is measured using a patch-clamp amplifier at a preset sampling rate until a predetermined time is reached, at which point the preset number of ion current values ​​are obtained.

7. The method according to claim 1, characterized in that, The step of dividing the preset number of ion current values ​​into multiple sets of current values ​​includes: The preset number of ion current values ​​are written into a file with a preset format to obtain a current data file; The current data file is split multiple times to obtain multiple file segments; The multiple current value sets are formed by using the ion current values ​​contained in each of the multiple file fragments.

8. An apparatus for detecting via events, characterized in that, It includes a data acquisition unit, a partitioning unit, and a determination unit; among which, The acquisition unit is used to measure the ion current between the first cavity and the second cavity to obtain a preset number of ion current values; wherein, the first cavity is filled with a base sequence; The dividing unit is used to divide the preset number of ion current values ​​into multiple current value sets; The determining unit is used to extract the peak values ​​of the plurality of current value sets respectively, and determine the plurality of current peak values ​​corresponding to the plurality of current value sets; The determining unit is further configured to: acquire the time points corresponding to each of the plurality of current peak values; determine the stable current time range corresponding to each of the plurality of current value sets based on the time points, a first time threshold, and a second time threshold; extract a plurality of candidate current values ​​corresponding to the plurality of current value sets based on the stable current time range; calculate the mean of the plurality of candidate current values ​​to obtain the mean of each of the plurality of current value sets; and determine the mean of each of the plurality of current value sets as the baseline current value corresponding to each of the plurality of current value sets. The determining unit is further configured to determine the current amplitude corresponding to each of the plurality of current peaks based on the plurality of current peaks and the baseline current values ​​corresponding to each of the plurality of current value sets; The determining unit is further configured to sort the current amplitudes corresponding to the plurality of current peaks to obtain a sorting result; determine the first quantile and the third quantile from the sorting result; subtract the first quantile and the third quantile to obtain a difference result; expand the difference result to obtain an expanded difference result; and sum the expanded difference result and the third quantile to obtain a detection threshold. The determining unit is further configured to determine that a via event has occurred in the base sequence when the current amplitude corresponding to each of the plurality of current peaks is greater than the detection threshold.

9. An electronic device, characterized in that, include: Memory, used to store executable instructions; A processor, when executing executable instructions stored in the memory, implements the method according to any one of claims 1 to 7.

10. A computer-readable storage medium, characterized in that, It stores executable instructions for causing a processor to execute, thereby implementing the method of any one of claims 1 to 7.

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