JP6740941B2 - Electrophoresis measurement method, data processing device, and data processing program - Google Patents

Description

The present invention relates to an electrophoretic measurement method that performs measurement by electrophoresis, and a data processing device and a data processing program that perform data processing on detection data obtained by electrophoresis.

When the measurement target substance is measured by electrophoresis, a measurement target sample in which a marker substance is mixed with the measurement target substance may be electrophoresed. As the marker substance, for example, a lower limit marker substance containing a component having an electrophoretic velocity faster than the component contained in the measurement target substance, and an upper limit marker substance containing a component having an electrophoretic velocity slower than the component contained in the measurement target substance Can be illustrated. On the other hand, as a method that does not use a marker substance, for example, a method using a dynamic time warping method (DTW: Dynamic Time Warping) as exemplified in Patent Document 1 below, a method using a dynamic programming method, and the like are available. Are known.

FIG. 8 is a diagram showing an example of detection data obtained by a method using a marker substance. In FIG. 8, the reference data D101 based on the detection data obtained by electrophoresing the reference sample containing the reference substance (known substance) and the measurement target sample containing the measurement target substance (unknown substance) are electrophoresed. Measurement target data D102 based on the obtained detection data is shown. The reference substance is a known reference substance, and the component contained in the measurement target substance can be identified by comparing the reference data D101 obtained from this reference substance with the measurement target data D102 obtained from the measurement target substance. ..

In addition to the reference substance, the lower limit marker substance and the upper limit marker substance are mixed in the reference sample. In the reference data D101, a peak P111 due to the detection of the lower limit marker substance component and a peak P112 due to the detection of the upper limit marker substance component appear. Further, between these peaks P111 and P112, a peak group P113 appears due to the detection of the component contained in the reference substance.

Similarly, the measurement target sample is mixed with the lower limit marker substance and the upper limit marker substance in addition to the measurement target substance. In the measurement target data D102, a peak P121 due to the detection of the lower limit marker substance component and a peak P122 due to the detection of the upper limit marker substance component appear. In addition, between these peaks P121 and P122, a peak group P123 appears due to the detection of the component contained in the measurement target substance.

In this example, it is obtained by electrophoresis so that the time width between the peak P111 and the peak P112 in the reference data D101 and the time width between the peak P121 and the peak P122 in the measurement target data D102 are constant. The detection data is standardized. That is, with reference to the peaks P111 and P121 corresponding to the lower limit marker substance in each data D101 and D102, the peaks P121 and P122 corresponding to the upper limit marker substance are standardized so as to have a constant time width. As a result, as shown in FIG. 8, the peaks P111 and P121 corresponding to the lower limit marker substance and the peaks P121 and P122 corresponding to the upper limit marker substance in the respective data D101 and D102 are in a state of being overlapped with each other.

By comparing the standard data D101 standardized in this way and the measurement target data D102, the components contained in the measurement target substance are identified based on the similarity of the peak groups P113 and P123 in each data D101 and D102. can do.

Japanese Patent No. 4957152

However, when comparing the respective data D101 and D102 standardized using the lower limit marker substance and the upper limit marker substance, even if the data is the same substance as the reference substance as the measurement target substance, as in the example of FIG. , The peak positions of the peak groups P113 and P123 corresponding to the same component may shift. The cause of this is considered to be the influence of the state of the device (microchip or the like) used for electrophoresis.

As described above, when the peak positions of the peak groups P113 and P123 are deviated, it becomes difficult to compare the data D101 and D102 (determine the degree of similarity). As a result, it may be difficult to identify the component contained in the measurement target substance, or in some cases, an accurate identification result may not be obtained.

The present invention has been made in view of the above circumstances, and provides an electrophoretic measurement method, a data processing device, and a data processing program capable of easily and accurately determining the degree of similarity between reference data and measurement target data. The purpose is to

(1) The electrophoresis measurement method according to the present invention includes a first electrophoresis step, a reference data acquisition step, a second electrophoresis step, a measurement target data acquisition step, and a data correction step. In the first electrophoresis step, a reference substance, a lower limit marker substance containing a component having an electrophoretic velocity higher than that of the component contained in the reference substance, and a component having an electrophoretic velocity slower than that of the component contained in the reference substance are selected. Detection data is obtained by subjecting a reference sample mixed with the upper limit marker substance containing it to electrophoresis. In the reference data acquisition step, the detection data acquired in the first electrophoresis step is normalized to a certain time width with reference to each peak of the lower limit marker substance and the upper limit marker substance included in the detection data. By doing so, the reference data is acquired. In the second electrophoresis step, the substance to be measured, the lower limit marker substance containing a component having an electrophoretic velocity faster than the component contained in the substance to be measured, and the electrophoretic velocity higher than the component contained in the substance to be measured. The detection data is obtained by subjecting the sample to be measured mixed with the upper limit marker substance containing a slow component to electrophoresis. In the measurement target data acquisition step, the detection data acquired by the second electrophoresis step is set to the constant time width with reference to each peak of the lower limit marker substance and the upper limit marker substance included in the detection data. The measurement target data is acquired by normalizing. In the data correction step, the reference data and the measurement target data are relatively expanded or contracted or shifted in the time axis direction.

With such a configuration, the reference data and the measurement target data are relatively expanded or contracted or shifted in the time axis direction, so that the data can be corrected to be easily compared with the reference data. That is, the peak group due to the detection of the component contained in the reference substance and the peak group due to the detection of the component contained in the measurement target substance are arranged so that the positions can be easily compared in the time axis direction. Can be corrected. Therefore, the degree of similarity between the reference data and the measurement target data can be easily and accurately determined. In particular, the influence of the difference in the components contained in the measurement target substance can be determined without being influenced by the electrophoresis device and the shift in the time axis direction due to the electrophoresis conditions.

(2) The data correction step may include a cross-correlation calculation step and a shift correction step. In the cross-correlation calculation step, the measurement target data is gradually shifted in the time axis direction, and the cross correlation between the measurement target data and the reference data at each shift amount is calculated. In the shift correction step, the measurement target data is shifted by a shift amount that maximizes the cross-correlation calculated in the cross-correlation calculation step.

According to such a configuration, based on the cross-correlation between the measurement target data and the reference data in each shift amount when the measurement target data is gradually shifted in the time axis direction, the shift that facilitates comparison with the reference data The data to be measured can be shifted by the amount.

(3) In the first electrophoresis step and the second electrophoresis step, detection data may be acquired by sampling at a predetermined reference cycle. In this case, at least one of the measurement target data before or after being stepwise shifted in the cross-correlation calculation step, or at least one of the reference data may be resampled at a setting cycle different from the reference cycle. ..

(4) The data correction step may include a first expansion/contraction data acquisition step, a first correlation coefficient calculation step, and an expansion/contraction direction determination step. In the first expansion/contraction data acquisition step, two expansion/contraction data are expanded based on the measurement target data when the measurement target data is expanded and contracted in the time axis direction, and when the expansion and contraction are performed, and the measurement target data when the expansion and contraction are performed. To get. In the first correlation coefficient calculating step, a correlation coefficient between the two expansion/contraction data and the reference data and a correlation coefficient between the measurement target data before expansion/contraction and the reference data are calculated. In the expansion/contraction direction determination step, the expansion/contraction direction of the measurement target data on the time axis is determined based on each correlation coefficient calculated in the first correlation coefficient calculation step.

According to such a configuration, the correlation coefficient between the two expansion/contraction data when the measurement target data is expanded and contracted in the time axis direction and the reference data, and the phase between the measurement target data before expansion and the reference data. Based on the number of relations, it is possible to accurately determine the expansion/contraction direction of the measurement target data that facilitates comparison with the reference data.

(5) In the first correlation coefficient calculation step, the two expansion/contraction data are stepwise shifted in the time axis direction, and the cross-correlation between the two expansion/contraction data and the reference data at each shift amount is calculated. Thereby, the correlation coefficient between the two expansion/contraction data shifted by the shift amount when the calculated cross-correlation becomes maximum and the reference data is calculated, and the measurement target data before expansion/contraction is plotted on the time axis. By stepwise shifting in the direction, by calculating the cross-correlation between the measurement target data and the reference data in each shift amount, the shifted by the shift amount when the calculated cross-correlation is maximum A correlation coefficient between the measurement target data and the reference data may be calculated.

(6) In the first electrophoresis step and the second electrophoresis step, detection data may be acquired by sampling at a predetermined reference cycle. In this case, at least one of the two expansion/contraction data and the measurement target data before the expansion/contraction, or the reference data acquired in the first expansion/contraction data acquisition step is resampled at a setting cycle different from the reference cycle. May be.

(7) The data correction step may include a second expansion/contraction data acquisition step, a second correlation coefficient calculation step, and an expansion/contraction correction step. In the second expansion/contraction data acquisition step, the measurement target data is expanded or shortened stepwise in the time axis direction, and a plurality of expansion/contraction data is acquired based on the measurement target data in each expansion/contraction amount. In the second correlation coefficient calculation step, a correlation coefficient between the plurality of expansion/contraction data and the reference data and a correlation coefficient between the measurement target data before expansion/contraction and the reference data are calculated. In the expansion/contraction correction step, the expansion/contraction data having the maximum correlation coefficient calculated in the second correlation coefficient calculation step or the measurement target data before expansion/contraction is used as the corrected measurement target data.

According to such a configuration, the correlation coefficient between the measurement target data and the reference data at each expansion/contraction amount when the measurement target data is expanded or shortened stepwise in the time axis direction, and the measurement target data before expansion/contraction. Based on the correlation coefficient between the reference data and the reference data, the data that can be easily compared with the reference data can be the corrected measurement target data.

(8) In the second correlation coefficient calculation step, the plurality of expansion/contraction data are stepwise shifted in the time axis direction, and the cross-correlation between the plurality of expansion/contraction data and the reference data at each shift amount is calculated. By calculating the correlation coefficient between the plurality of expansion/contraction data and the reference data shifted by the shift amount when the calculated cross-correlation becomes maximum, the measurement target data before expansion/contraction is plotted on the time axis. By stepwise shifting in the direction, by calculating the cross-correlation between the measurement target data and the reference data in each shift amount, the shifted by the shift amount when the calculated cross-correlation is maximum A correlation coefficient between the measurement target data and the reference data may be calculated.

(9) In the first electrophoresis step and the second electrophoresis step, the detection data may be acquired by sampling at a predetermined reference cycle. In this case, at least one of the plurality of expansion/contraction data and the measurement target data before expansion or expansion, which is acquired in the second expansion/contraction data acquisition step, or the reference data is resampled at a setting cycle different from the reference cycle. May be.

(10) The data correction step may include an expansion/contraction correction step and a shift correction step. In the expansion/contraction correction step, the measurement target data is expanded or contracted in the time axis direction. In the shift correction step, the measurement target data is shifted in the time axis direction before or after the expansion/contraction correction step.

(11) In at least one of the reference data acquisition step and the measurement target data acquisition step, the reference data or the measurement target data is obtained by multiplying the detection intensity in each cycle of the detection data resampled at the set cycle by a coefficient. May be obtained.

According to such a configuration, a coefficient is added to the detection intensity in at least one of the peak group due to the detection of the component contained in the reference substance and the peak group due to the detection of the component contained in the measurement target substance. By performing the multiplication, it is possible to obtain reference data or measurement target data that enables more accurate comparison for each component.

(12) In the first electrophoresis step, a plurality of detection data corresponding to each reference sample may be acquired by subjecting a plurality of reference samples containing different reference substances to electrophoresis. In the reference data acquisition step, a plurality of reference data may be acquired from the plurality of detection data acquired in the first electrophoresis step. In the data correction step, the measurement target data may be relatively expanded or contracted or shifted in the time axis direction for the plurality of reference data. In this case, the electrophoretic measurement method is based on a determination result in the similarity determination step of determining the similarity between the reference data and the measurement target data that have been relatively expanded or contracted or shifted, and the determination result in the similarity determination step. It may further include a matching data selecting step of selecting matching data from the plurality of reference data.

According to such a configuration, it is possible to determine the similarity between the plurality of reference data and the measurement target data, and select the matching data that matches the measurement target data from the plurality of reference data based on the determination result. You can

(13) In the similarity determination step, the similarity may be determined based on the correlation coefficient between each reference data and the measurement target data, and the peak area of the measurement target data.

According to such a configuration, the similarity is determined by using the peak area of the measurement target data in addition to the correlation coefficient between each reference data and the measurement target data. Therefore, the degree of similarity can be determined more accurately by considering not only the peak position but also the peak area.

(14) In the similarity determination step, the similarity is calculated based on the correlation coefficient between each reference data and the measurement target data, and the area ratio of the peak area of the measurement target data to the peak area of each reference data. You may judge.

According to such a configuration, the similarity is determined by using the area ratio of the peak area of the measurement target data to the peak area of each reference data, in addition to the correlation coefficient between each reference data and the measurement target data. Therefore, the degree of similarity can be determined more accurately by considering not only the peak position but also the area ratio of the peak areas.

(15) In the similarity determination step, a correlation coefficient between each reference data and the measurement target data, a peak area of the measurement target data, and an area of a peak area of the measurement target data with respect to a peak area of each reference data. The degree of similarity may be determined based on the ratio.

According to such a configuration, in addition to the correlation coefficient between each reference data and the measurement target data, the peak area of the measurement target data, and the area ratio of the peak area of the measurement target data to the peak area of each reference data The degree of similarity is determined by using. Therefore, not only the peak position but also the peak area and the area ratio are considered, and the similarity can be determined more accurately.

(16) As the correlation coefficient between each reference data and the measurement target data, an average value of the correlation coefficients calculated in each of the plurality of regions included in the certain time width may be used.

With such a configuration, the similarity can be determined more accurately by using the average value of the correlation coefficients calculated in each of the plurality of regions.

(17) A data processing device according to the present invention is a data processing device that performs data processing using reference data and measurement target data, and relatively expands or contracts the reference data and the measurement target data in a time axis direction. Alternatively, a data correction unit for shifting is provided.

(18) A data processing program according to the present invention is a data processing program that performs data processing using reference data and measurement target data, and relatively expands or contracts the reference data and the measurement target data in the time axis direction. Alternatively, the computer is made to function as a data correction unit for shifting.

According to the present invention, by expanding or contracting the reference data and the measurement target data relatively in the time axis direction, it is possible to correct the data to be easily compared with the reference data. The degree of similarity can be easily and accurately determined.

It is a block diagram showing an example of an electrophoresis system which performs measurement using an electrophoresis measurement method concerning one embodiment of the present invention. It is the flowchart which showed an example of the electrophoresis measurement method. It is a figure showing an example of corrected measurement object data. It is the flowchart which showed an example of the specific process at the time of determining the expansion-contraction direction of measurement object data. It is the flowchart which showed an example of the specific process at the time of expanding/contracting measurement object data. It is the flowchart which showed an example of the specific process at the time of shifting measurement object data. It is a figure showing an example of standard data and measurement object data after amendment. It is the figure which showed an example of the detection data obtained by the method of using a marker substance.

1. Configuration of Electrophoresis System FIG. 1 is a block diagram showing an example of an electrophoresis system that performs measurement using an electrophoresis measurement method according to an embodiment of the present invention. The electrophoresis system includes an electrophoresis device 1 and a data processing device 2.

In the electrophoretic device 1, for example, a microchip (not shown) in which a channel is formed inside a plate-shaped member is used, and the sample is electrophoresed in the channel. The electrophoretic device 1 is provided with a detection unit 11 having, for example, a light emitting unit and a light receiving unit. At the time of measurement, excitation light is emitted from the light emitting unit of the detection unit 11 to the sample electrophoresed in the channel, and the fluorescence from the sample can be detected by the light receiving unit. However, the detection unit 11 is not limited to the configuration that detects the fluorescence from the sample, and may have a configuration that detects the absorbance by receiving the transmitted light from the sample, for example.

The data processing device 2 processes data (detection intensity of absorbance) input from the detection unit 11 of the electrophoretic device 1. The data processing device 2 is configured to include, for example, a CPU (Central Processing Unit), and the CPU executes a program so that the data acquisition unit 21, the data correction unit 22, the similarity determination unit 23, the matching data selection unit 24, and the like. Function as.

The data acquisition unit 21 acquires data input from the detection unit 11 of the electrophoretic device 1. The data correction unit 22 performs a process of correcting the data acquired by the data acquisition unit 21. The data correction unit 22 includes, for example, a cross-correlation calculation unit 221, a shift correction unit 222, a stretch data acquisition unit 223, a correlation coefficient calculation unit 224, a stretch direction determination unit 225, and a stretch correction unit 226. The similarity determination unit 23 performs a process of determining the similarity by comparing the data (waveform data) with each other. The matching data selecting unit 24 performs a process of selecting any data as matching data based on the determination result by the similarity determining unit 23.

2. Specific Example of Electrophoresis Measurement Method FIG. 2 is a flowchart showing an example of the electrophoresis measurement method. In the present embodiment, the data obtained by measuring the reference sample and the sample to be measured by the electrophoresis device 1 are processed by the data processing device 2, respectively.

The reference sample is a sample in which the reference substance, the lower limit marker substance and the upper limit marker substance are mixed. The reference substance is a known reference substance. The lower limit marker substance is a component having an electrophoretic velocity faster than the component contained in the reference substance, and the upper limit marker substance is a component having an electrophoretic velocity slower than the component contained in the reference substance. Such reference sample is electrophoresed by the electrophoresis apparatus 1, and the data input from the detection unit 11 is sampled by the data acquisition unit 21 to acquire detection data based on the reference sample (step S101: first). Electrophoresis process). At this time, the sampling by the data acquisition unit 21 is performed in a predetermined cycle (reference cycle).

In the detection data obtained based on the reference sample, in addition to the peak group resulting from the detection of the components contained in the reference substance, the lower limit marker substance component was detected so that the peak group was sandwiched in the time axis direction. And a peak due to detection of a component of the upper limit marker substance. Such detection data is normalized by the data acquisition unit 21 to have a constant time width with reference to each peak of the lower limit marker substance and the upper limit marker substance (step S102).

Specifically, as in the reference data D101 of FIG. 8, the detection data is obtained by using a migration index such that the position of the peak P111 of the lower limit marker substance is 0% and the position of the peak P112 of the upper limit marker substance is 100%. Is standardized to a fixed time width (0-100% exponential width). It is assumed that the time axis in the present embodiment includes axes of other parameters such as the migration index calculated on the basis of time.

At this time, the data acquisition unit 21 resamples the standardized detection data at a cycle (setting cycle) set to a value different from the reference cycle, thereby acquiring the reference data D101 as shown in FIG. (Step S103: reference data acquisition step). The set cycle is set to be longer than the reference cycle, for example. When re-sampling the standardized detection data in the set cycle, an interpolation value calculated by using spline interpolation or polynomial interpolation is added or subtracted to/from each detection intensity in the reference cycle. As a result, the respective detection intensities in the set cycle are interpolated.

Although only one reference data D101 may be acquired, in the present embodiment, a plurality of reference data D101 is acquired using a plurality of reference samples. That is, in the first electrophoresis step of step S101, a plurality of reference samples containing different reference substances are electrophoresed to obtain a plurality of detection data corresponding to each reference sample. Further, in the reference data acquisition step of step S103, a plurality of reference data are acquired from the plurality of detection data acquired in the first electrophoresis step. The lower limit marker substance and the upper limit marker substance contained in the reference sample are the same in each reference sample.

The measurement target sample is a sample in which the measurement target substance, the lower limit marker substance and the upper limit marker substance are mixed. The substance to be measured is a substance to be measured and includes, for example, DNA and RNA. The lower limit marker substance has a higher electrophoretic velocity than the component contained in the measurement target substance, and the upper limit marker substance has a lower electrophoretic velocity than the component contained in the measurement target substance, and is mixed with the measurement target substance. The lower limit marker substance and the upper limit marker substance and the lower limit marker substance and the upper limit marker substance mixed with the reference substance are the same substance. Such a measurement target sample is electrophoresed by the electrophoresis apparatus 1, and the data input from the detection unit 11 is sampled by the data acquisition unit 21 to acquire the detection data based on the measurement target sample (step S104: Second electrophoresis step). At this time, the sampling by the data acquisition unit 21 is performed in the same cycle (reference cycle) as when the detection data based on the reference sample is acquired (step S101).

In the detection data obtained based on the measurement target sample, in addition to the peak group due to the detection of the components contained in the measurement target substance, the components of the lower limit marker substance are sandwiched between the peak groups in the time axis direction. A peak due to detection and a peak due to detection of a component of the upper limit marker substance are included. Such detection data is normalized by the data acquisition unit 21 to have a constant time width with reference to the respective peaks of the lower limit marker substance and the upper limit marker substance (step S105).

Specifically, detection is performed using a migration index such that the position of the peak P121 of the lower limit marker substance is 0% and the position of the peak P122 of the upper limit marker substance is 100% as in the measurement target data D102 of FIG. The data is normalized to a fixed time width (0-100% exponential width). At this time, the data acquisition unit 21 resamples the standardized detection data at the above-mentioned setting cycle to acquire measurement target data D102 as shown in FIG. 8 (step S106: measurement target data acquisition step). .. When re-sampling the standardized detection data in the set cycle, an interpolation value calculated by using spline interpolation or polynomial interpolation is added to or subtracted from each detection intensity in the reference cycle. As a result, the respective detection intensities in the set cycle are interpolated.

In this way, when the reference data D101 is acquired (step S103) and the measurement target data D102 is acquired (step S106), by re-sampling at the same cycle (setting cycle), at the same timing. It is possible to compare the sampled data with each other. Thereafter, the data correction unit 22 expands or shifts the measurement target data D102 in the time axis direction (migration index axis direction) with respect to the reference data D101, thereby correcting the measurement target data (steps S107 to S109). : Data correction process).

Specifically, first, the direction (expansion/contraction direction) of extending or shortening the measurement target data on the time axis (migration index axis) is determined (step S107). Then, after the measurement target data is expanded or shortened in the determined expansion/contraction direction (step S108), the expansion/contraction measurement target data is shifted in the time axis direction (step S109). However, as will be described later, the measurement target data may not be expanded or shortened.

In the present embodiment, the data correction process of steps S107 to S109 is performed using each reference data D101. That is, in the data correction step, the measurement target data D102 is relatively expanded or contracted or shifted in the time axis direction for the plurality of reference data D101. As a result, it is possible to correct the measurement target data to be easily compared with each reference data.

After that, the similarity determination unit 23 determines each similarity by comparing each reference data D101 with the measurement target data corrected using each reference data D101 (step S110: similarity determination step). ). Then, based on the determination result, the matching data selecting unit 24 selects matching data from the plurality of reference data D101 (step S111: matching data selecting step). That is, the reference data D101 having the highest degree of similarity to the corrected measurement target data is selected as the matching data.

3. Example of Corrected Measurement Target Data FIG. 3 is a diagram showing an example of the corrected measurement target data D103. In this example, the measurement target data D102 shown in FIG. 8 is expanded and contracted and shifted in the time axis direction with reference to the reference data D101, and the corrected measurement target data D103 is obtained. ..

In the corrected measurement target data D103, the measurement target data D102 is shortened so that the interval in the time axis direction between the peak P121 of the lower limit marker substance and the peak P122 of the upper limit marker substance is narrowed. Then, by shifting the shortened measurement target data D102 in the time axis direction, as shown in FIG. 3, the position in the time axis direction of the peak group P123 due to the detection of the component contained in the measurement object substance is determined. The measurement target data D103 corrected so as to approach the position of the peak group P113 in the time axis direction due to the detection of the component contained in the reference substance is obtained.

As described above, in the present embodiment, the measurement target data D102 is expanded or contracted and shifted in the time axis direction with reference to the reference data D101, so that the measurement target data D103 can be easily compared with the reference data D101. .. That is, the peak group P113 resulting from the detection of the component contained in the reference substance and the peak group P123 resulting from the detection of the component contained in the measurement target substance are positioned so as to be easily compared in the time axis direction. The measurement target data D102 can be corrected. Therefore, the degree of similarity between the reference data D101 and the measurement target data D103 can be easily and accurately determined.

4. Processing when deciding expansion/contraction direction FIG. 4 is a flowchart showing an example of specific processing when deciding the expansion/contraction direction of the measurement target data D102. This processing is performed using each of the plurality of reference data D101, and the expansion/contraction direction of the measurement target data D102 corresponding to each reference data D101 is determined.

When determining the expansion/contraction direction of the measurement target data D102, first, the expansion/contraction data acquisition unit 223 expands and contracts the measurement target data D102 in the time axis direction, and measures the contracted measurement target data and the contracted data. The measurement target data of is acquired (steps S201 and S202: first expansion/contraction data acquisition step). As a result, three pieces of data including the expanded measurement target data, the shortened measurement target data, and the measurement target data D102 before expansion/contraction are acquired. At this time, the expanded measurement object data, the shortened measurement object data, and the measurement object data D102 before expansion and contraction are respectively resampled at a predetermined set cycle.

Specifically, the peak P121 of the lower limit marker substance or the peak P122 of the upper limit marker substance is moved by an arbitrary amount along the time axis direction. At this time, if the peak P121 of the lower limit marker substance is moved in the plus direction (direction of increasing migration index) or the peak P122 of the upper limit marker substance is moved in the minus direction (direction of decreasing migration index), it is elongated. The data to be measured can be obtained. On the other hand, if the peak P121 of the lower limit marker substance is moved in the negative direction or the peak P122 of the upper limit marker substance is moved in the positive direction, shortened measurement target data is obtained.

After that, the correlation coefficient calculation unit 224 shifts the expanded measurement target data, the shortened measurement target data, and the measurement target data D102 before expansion respectively stepwise in the time axis direction, and the respective shift amounts. The cross-correlation between each measurement target data and the reference data D101 is calculated (steps S203 and S204). Then, for each of the expanded measurement target data, the shortened measurement target data, and the measurement target data D102 before expansion and contraction, the shift amount that maximizes the calculated cross-correlation is determined, and each measurement target data is calculated. The shift amount is shifted by the determined shift amount (step S205). The specific processing for shifting the measurement target data and the specific processing for calculating the cross-correlation are the same as the processing described later with reference to FIG. 6, and thus detailed description will be omitted.

The correlation coefficient calculating unit 224 calculates the correlation coefficient (R+) between the shifted expanded measurement target data and the reference data D101, and the correlation coefficient (R+) between the shifted shortened measurement target data and the reference data D101. -) and the correlation coefficient (R0) between the shifted measurement target data before expansion and the reference data D101 are calculated (step S206: first correlation coefficient calculation step). The expansion/contraction direction determination unit 225 determines the expansion/contraction direction of the measurement target data D102 on the time axis based on the three types of correlation coefficients R+, R−, R0 calculated in this way (step S207: expansion/contraction direction determination). Process).

A detailed description of the correlation coefficient is omitted because it is well known, but it is a value indicating how many peaks appear at the same position. The closer the peak positions of the data to be compared are to “1”. The values are close to each other, and the farther the position of the peak is, the closer to "-1". Therefore, the expansion/contraction direction of the measurement target data D102 can be determined using the following criteria (A) to (D).
(A): When correlation coefficient R0>correlation coefficient R+ and correlation coefficient R0>correlation coefficient R−, it is determined that measurement target data D102 is “not expanded/contracted” (B): other than (A) And correlation coefficient R+>correlation coefficient R−, it is determined that measurement target data D102 is “expanded” (C): other than (A), and correlation coefficient R−> In the case of the correlation coefficient R+, the measurement target data D102 is determined to be “shortened” (D): in the cases other than (A) to (C), the peak P121 of the lower limit marker substance or the peak of the upper limit marker substance While increasing the movement amount of P122 in the time axis direction, steps S201 to S206 are repeated until one of (A) to (C) is satisfied.

According to the processing as shown in FIG. 4, the correlation coefficients R+ and R− between the two expansion/contraction data and the reference data D101 when the measurement target data D102 is expanded and contracted in the time axis direction, and the measurement target Based on the correlation coefficient R0 between the data D102 and the reference data D101, it is possible to accurately determine the expansion/contraction direction of the measurement target data D102 that is easy to compare with the reference data D101.

5. Processing during expansion/contraction FIG. 5 is a flowchart showing an example of specific processing when expanding/compressing the measurement target data D102. This processing is performed using each of the plurality of reference data D101, so that the measurement target data D102 expanded and contracted corresponding to each reference data D101 is obtained.

When the expansion/contraction direction of the measurement target data D102 is determined, the expansion/contraction data acquisition unit 223 gradually expands or shortens the measurement target data D102 in the time axis direction according to the expansion/contraction direction. That is, if it is determined in the step S207 of FIG. 4 that the measurement target data D102 is to be expanded, the measurement target data D102 is expanded stepwise in the time axis direction, and if it is determined to be shortened, the measurement is performed. The target data D102 is shortened stepwise in the time axis direction. Then, a plurality of expansion/contraction data are acquired by re-sampling the respective measurement target data when expanded or shortened in stages at a predetermined setting cycle (step S301: second expansion/contraction data acquisition step). If it is determined in step S207 in FIG. 4 that the measurement target data D102 is not expanded or contracted, the step S301 is not performed.

Specifically, when it is determined to extend the measurement target data D102, the amount of movement of the lower limit marker substance peak P121 in the plus direction or the amount of movement of the upper limit marker substance peak P122 in the minus direction is allowed. It is increased stepwise within the range of expansion and contraction amount, and the measurement target data in each expansion amount is created. At this time, the measurement target data before expansion/contraction has the expansion amount of zero and is included in the expansion data of multiple stages. On the other hand, when it is determined that the measurement target data D102 is to be shortened, the amount of movement of the lower limit marker substance peak P121 in the negative direction or the amount of movement of the upper limit marker substance peak P122 in the positive direction is the allowable expansion/contraction amount. It is increased stepwise within the range, and the measurement target data at each shortening amount is created. At this time, the measurement target data before expansion/contraction is included in the shortened data in a plurality of steps, with the shortening amount being zero. Each stepwise decompressed data or shortened data is resampled at a predetermined cycle.

When a plurality of pieces of expansion/contraction data corresponding to a plurality of levels of expansion/contraction are acquired in this way, the correlation coefficient calculation unit 224 shifts each of the plurality of levels of expansion/contraction data stepwise in the time axis direction, and each expansion/contraction is performed. The cross-correlation between each measurement target data and the reference data D101 at each shift amount of the stage is calculated (steps S302 and S303). Then, for the measurement target data of each expansion/contraction stage, the shift amount that maximizes the cross-correlation calculated from each shift amount is determined, and the measurement target data of each expansion/contraction stage is shifted by the determined shift amount ( Step S304). The specific processing for shifting the measurement target data and the specific processing for calculating the cross-correlation are the same as the processing described later with reference to FIG. 6, and thus detailed description will be omitted.

The correlation coefficient calculation unit 224 calculates the correlation coefficient between the reference data D101 and the data after being shifted by the shift amount determined in S304 for the measurement target data at each expansion/contraction stage (step S305: second phase). Relation number calculation process). That is, the correlation coefficient between the resampled measurement target data D102 and the reference data D101 at each expansion/contraction amount (including the case where the expansion/contraction is not performed) is calculated.

Then, the expansion/contraction correction unit 226 determines the data having the maximum calculated correlation coefficient among the plurality of expansion/contraction measurement target data, and sets it as the corrected measurement target data (step S306: expansion/contraction correction step). As a result, the data having the highest similarity to the reference data D101 is determined as the corrected measurement target data D102.

According to the processing as shown in FIG. 5, based on the correlation coefficient between the measurement target data and the reference data D101 at each expansion/contraction amount when the measurement target data D102 is expanded or shortened stepwise in the time axis direction. By adopting as the corrected measurement target data D103, it is possible to obtain the corrected measurement target data D103 having high identification accuracy with the measurement substance when compared with the reference data D101.

6. Processing at Shift FIG. 6 is a flowchart showing an example of specific processing at the time of shifting the measurement target data D102. This processing is performed using each of the plurality of reference data D101, so that the measurement target data D102 shifted corresponding to each reference data D101 is obtained.

When shifting the measurement target data D102, the cross-correlation calculation unit 221 shifts the measurement target data D102 (expanded measurement target data or non-expanded measurement target data) stepwise in the time axis direction, respectively. The cross-correlation between the measurement target data D102 and the reference data D101 at the shift amount is calculated (steps S401 and S402: cross-correlation calculation step).

Specifically, the measurement target data D102 is gradually shifted in the plus direction and the minus direction within the allowable shift amount range. At this time, the measurement target data D102 is shifted stepwise within a suitable range preset as a range in the time axis direction in which the peak group P123 of the measurement target data D102 appears. The applicable range is set to different values according to the type of the substance to be measured, and in the example of FIG. 3, it is set in multiple steps within the range of 55 to 96%. Note that the above-mentioned correlation coefficient is also calculated based on the data within the above-mentioned conformity range.

The cross-correlation is well known, and thus a detailed description thereof will be omitted. However, the signal strength of the measurement target data D102 and the signal strength of the reference data D101 obtained in the above-mentioned setting cycle are data sampled at the same timing. A value obtained by multiplying and multiplying the multiplied values becomes a cross-correlation. Therefore, the higher the degree of similarity between the compared data, the larger the value of the cross-correlation.

The shift correction unit 222 shifts the measurement target data D102 by the shift amount when the cross-correlation calculated as described above becomes maximum (step S403: shift correction step). As a result, the measurement target data D102 is shifted in the time axis direction by the shift amount having the highest similarity to the reference data D101. The measurement target data D102 is resampled at the above-mentioned setting cycle before or after being shifted stepwise in step S401.

According to the processing as shown in FIG. 6, the reference is calculated based on the cross-correlation between the measurement target data D102 and the reference data D101 at each shift amount when the measurement target data D102 is stepwise shifted in the time axis direction. The measurement target data D102 can be shifted by a shift amount that facilitates comparison with the data D101. By such processing of FIGS. 4 to 6, the corrected measurement target data D103 as shown in FIG. 3 is obtained.

7. Modification Example of Expansion/Contraction Correction and Shift Correction Not limited to the above embodiment, the expansion/contraction correction step of expanding or contracting the measurement target data D102 in the time axis direction, and the measurement target data D102 before or after the expansion/contraction correction step. The following configuration is also possible as long as the configuration includes a shift correction step of shifting in the time axis direction.

For example, without first determining the expansion/contraction direction, the measurement target data D102 is expanded/contracted stepwise in the time axis direction within a predetermined expansion/contraction range and expansion/contraction step (expansion/contraction correction step). The expansion/contraction range is, for example, 95% to 105%, and the expansion/contraction step is, for example, 1%. As a result, eleven pieces of measurement target data D102 including data that has not been expanded or contracted are obtained. The shift correction process is performed on the plurality of measurement target data D102 thus obtained.

Specifically, the plurality of measurement target data D102 are shifted stepwise in the time axis direction, and the cross-correlation between the measurement target data D102 and the reference data D101 at each shift amount is calculated. Then, for each of the plurality of measurement target data D102, the measurement target data D102 (optimal shift data) shifted by the shift amount at which the calculated cross-correlation becomes maximum is obtained. The correlation coefficient between the plurality of optimum shift data thus obtained and the reference data D101 is calculated, and the optimum shift data (measurement target data D102) having the maximum calculated correlation coefficient is the measurement target after correction. The data is D103.

However, the configuration is not limited to the configuration in which the shift correction process is performed after the expansion/contraction correction process as described above, and the shift correction process may be performed before the expansion/contraction correction process. Further, when determining the expansion/contraction direction, first, two expansion/contraction data (for example, 99% and 101%) are obtained, and the correlation coefficient between these two expansion/contraction data and the reference data D101 and the pre-expansion and contraction data By calculating the correlation coefficient between the measurement target data (100%) and the reference data D101, the expansion/contraction direction can be determined from the one having the maximum correlation coefficient.

8. First Example of Similarity Determining Hereinafter, a specific mode in which the similarity determining unit 23 determines the similarity will be described. The similarity determination unit 23 determines the correlation coefficient A between each reference data D101 and the corrected measurement target data D103, the peak area B of the corrected measurement target data D103, and the corrected measurement of the peak area of each reference data D101. The degree of similarity is determined using the area ratio C of the peak areas of the target data D103.

FIG. 7 is a diagram showing an example of the reference data D101 and the measurement target data D103 after correction. As shown in FIG. 7, when the peak position of the reference data D101 and the peak position of the corrected measurement target data D103 are close to each other, the correlation coefficient A has a value close to “1”. Therefore, it is also possible to select the reference data having the largest correlation coefficient A from the plurality of reference data based on only the correlation coefficient A, and use the reference data as the matching data that matches the measurement target data. .. However, in this case, since the signal intensity of the reference data D101 and the signal intensity of the corrected measurement target data D103 are not taken into consideration, the determination result is the same even when the signal intensities of both are significantly different, It may not be possible to accurately determine whether the data is appropriate.

Therefore, in the present embodiment, in addition to the correlation coefficient A between each reference data D101 and the measurement target data D103, the peak area B of the measurement target data D103 and the peak of the measurement target data D103 with respect to the peak area of each reference data D101. The area ratio C of the areas is also used to determine the similarity. Thereby, not only the peak position but also the peak area B and the area ratio C are considered, and the similarity can be determined more accurately.

Specifically, a predetermined region (region of interest) on the time axis (including the migration index axis) is set, and the peak area B and the area ratio C within the region of interest are calculated. In the example of FIG. 7, a region having a migration index of 60 to 95% is set as the region of interest. The similarity determination unit 23 calculates an area surrounded by the time axis and the reference data D101 as a peak area of the reference data D101 within a preset region of interest, and is surrounded by the time axis and the measurement target data D103. The calculated area is calculated as the peak area of the measurement target data D103. Then, the area ratio C is calculated by dividing the calculated peak area of the measurement target data D103 by the peak area of the reference data D101.

The similarity determination unit 23 uses the calculated correlation coefficient A, peak area B, and area ratio C to obtain the evaluation value E1 by the following formula. Α, β, and γ in the following formula are coefficients, which are determined using known data (teacher data) measured in advance.
E1=A×α+B×β+C×γ

The compatible data selecting unit 24 selects the compatible data from the plurality of reference data D101 using the calculated evaluation value E1. At this time, the reference data D101 having the largest evaluation value E1, that is, the reference data D101 having the highest degree of similarity to the measurement target data D103 is selected as the matching data. However, when the largest evaluation value E1 is not equal to or larger than a predetermined threshold value, it may be determined that there is no matching data and no matching data may be selected.

In the above example, a configuration has been described in which the similarity determination unit 23 determines the similarity using the correlation coefficient A, the peak area B, and the area ratio C. However, the configuration is not limited to such a configuration, and the configuration may be such that the similarity is determined using only the correlation coefficient A and the peak area B, or the similarity is determined using only the correlation coefficient A and the area ratio C. It may be configured to determine the degree.

9. Second Example of Similarity Judgment When calculating the correlation coefficient A between each reference data D101 and the corrected measurement target data D103, the region of interest is divided into a plurality of regions in the time axis direction, and calculated in each region. The average value of the correlation coefficients may be calculated. For example, in FIG. 7, a region of interest having a migration index of 60 to 95% is divided into a first region of 60 to 75%, a second region of 75 to 87.5%, and a third region of 87.5 to 95%. .. Then, a correlation coefficient is calculated in each of the first to third areas, and an average value such as an arithmetic mean, a weighted average, or a geometric mean of the correlation coefficients is obtained, and the average value is calculated. Is calculated as

As described above, as the correlation coefficient A between the reference data D101 and the measurement target data D103, by using the average value of the correlation coefficients respectively calculated in the plurality of regions included in the constant time width, it is possible to more accurately The degree of similarity can be determined. However, not only the correlation coefficient A but also the peak area B or the area ratio C may be the average value of the peak areas or the area ratios calculated in a plurality of regions.

10. Other Modifications In the above embodiment, when the standardized detection data is resampled at the above-mentioned setting cycle in step S103 (reference data acquisition step) or step S106 (measurement target data acquisition step) of FIG. The case has been described where the detected intensities in the reference cycle are interpolated and the interpolated detected intensities are used as they are. However, the configuration is not limited to such a configuration, and the configuration may be such that the reference data D101 or the measurement target data D102 is acquired by multiplying each detection intensity in the reference cycle by a coefficient.

Specifically, each detected intensity is multiplied by a coefficient represented by the following formula (1). The reference sampling timing is set, for example, at the start point of the matching range.

The following formula (2-1) or formula (2-2) is used as the above formula (2).

In Expression (2-1), D is the effective migration length (μm), and T(i) is the migration time (s) corresponding to the i-th sampled timing. When the coefficient is calculated by the equation (1) using the equation (2-1), D is eliminated and becomes a function of only T(i) and T(i+1). As a result, it can be said that the calculated coefficient is the ratio of the difference in the moving distance after the observation.

On the other hand, in Expression (2-2), S is an arbitrary sampling timing (s), which is set, for example, 1 second after the start point of the matching range. D and T(i) are the same as in the case of the equation (2-1). When the coefficient is calculated by the equation (1) using the equation (2-2), D and S are eliminated and only the functions of T(i) and T(i+1) are obtained. As a result, it can be said that the calculated coefficient is the ratio of the difference in the moving distance after 1 second.

The calculation using the coefficient as described above may be performed only in either one of the reference data acquisition step and the measurement target data acquisition step. Thereby, the detection intensity is multiplied by a coefficient in at least one of the peak group P113 resulting from the detection of the component contained in the reference substance and the peak group P123 resulting from the detection of the component contained in the measurement target substance. With this, it is possible to acquire the reference data D101 or the measurement target data D102 that enables more accurate comparison for each component.

In steps S201 and S202 of FIG. 4, when the peak P121 of the lower limit marker substance or the peak P122 of the upper limit marker substance is moved along the time axis direction, the center of the compatible range after the movement is the same as before the movement. The measurement target data D102 may be shifted so as to be positioned. Similarly, in step S301 of FIG. 5, when the peak P121 of the lower limit marker substance or the peak P122 of the upper limit marker substance is moved along the time axis direction, the center of the fitted range after the movement is the same as before the movement. The measurement target data D102 may be shifted so as to be in the same position.

In the above embodiment, the configuration has been described in which the measurement target data D102 is expanded/contracted and then shifted. However, the configuration is not limited to such a configuration, and the configuration may be such that the measurement target data D102 is shifted and then expanded or contracted, or only the expansion or contraction or shift is performed on the measurement target data D102. It may be. Further, instead of expanding/contracting or shifting the measurement target data D102 in the time axis direction with reference to the reference data D101, the reference data D101 may be expanded/contracted in the time axis direction with reference to the measurement target data D102. That is, the reference data D101 and the measurement target data D102 may be configured to relatively expand or contract or shift in the time axis direction. In this case, the reference data D101 may be resampled at the set cycle.

In the above embodiments, the specific configuration of the data processing device 2 has been described, but it is also possible to provide a program (data processing program) for causing a computer to function as the data processing device 2. In this case, the above-mentioned program may be provided in a state of being stored in a storage medium, or may be provided so that the program itself is provided.

DESCRIPTION OF SYMBOLS 1 Electrophoresis device 2 Data processing device 11 Detection part 21 Data acquisition part 22 Data correction part 23 Similarity determination part 24 Matching data selection part 221 Cross correlation calculation part 222 Shift correction part 223 Expansion/contraction data acquisition part 224 Correlation coefficient calculation part 225 Stretching direction determination unit 226 Stretching correction unit D101 Standard data D102 Measurement target data D103 Measurement target data P111 Lower marker substance peak P112 Upper marker substance peak P113 Peak group P121 Lower marker substance peak P122 Upper marker substance peak P123 Peak group

Claims (18)

A reference substance, a lower limit marker substance containing a component having an electrophoretic rate faster than the component contained in the reference substance, and an upper limit marker substance containing a component having a lower electrophoretic rate than the component contained in the reference substance were mixed. A first electrophoresis step of obtaining detection data by subjecting a reference sample to electrophoresis,
The reference data is obtained by normalizing the detection data obtained by the first electrophoresis step to a certain time width with reference to each peak of the lower limit marker substance and the upper limit marker substance contained in the detection data. The reference data acquisition process to be acquired,
Measurement target substance, the lower limit marker substance containing a component having an electrophoretic velocity faster than the component contained in the measurement target substance, and the upper limit marker containing a component having an electrophoretic velocity slower than the component contained in the measurement target substance A second electrophoresis step in which detection data is obtained by subjecting a measurement target sample mixed with a substance to electrophoresis,
The detection data obtained by the second electrophoresis step is standardized to the constant time width with reference to the peaks of the lower limit marker substance and the upper limit marker substance contained in the detection data, to thereby obtain a measurement target. A measurement target data acquisition process for acquiring data,
And a data correction step of relatively expanding or contracting or shifting the reference data and the measurement target data in the time axis direction. In the data correction step,
A cross-correlation calculation step of calculating the cross-correlation between the measurement target data and the reference data in each shift amount by shifting the measurement target data stepwise in the time axis direction,
The electrophoretic measurement method according to claim 1, further comprising a shift correction step of shifting the measurement target data by a shift amount at which the cross correlation calculated by the cross correlation calculation step becomes maximum. In the first electrophoretic step and the second electrophoretic step, detection data is acquired by sampling at a predetermined reference cycle,
At least one of the measurement target data before or after being stepwise shifted in the cross-correlation calculation step, or at least one of the reference data is resampled at a setting cycle different from the reference cycle. Item 3. The electrophoretic measurement method according to Item 2. In the data correction step,
A first expansion/contraction data acquisition step of acquiring two expansion/contraction data based on the measurement target data when the measurement target data is expanded and contracted in the time axis direction, and the measurement target data when expanded and the measurement target data when contracted. When,
A first correlation coefficient calculation step of calculating a correlation coefficient between the two expansion/contraction data and the reference data, and a correlation coefficient between the measurement target data before expansion and the reference data,
The expansion/contraction direction determination step of determining the expansion/contraction direction of the measurement target data on the time axis based on each correlation coefficient calculated by the first correlation coefficient calculation step. The electrophoretic measurement method according to any one of 3 above. In the first correlation coefficient calculation step, the two expansion/contraction data are stepwise shifted in the time axis direction, and the cross-correlation between the two expansion/contraction data and the reference data in each shift amount is calculated, The correlation coefficient between the two expansion/contraction data and the reference data shifted by the shift amount when the calculated cross-correlation becomes maximum is calculated, and the measurement target data before expansion/contraction is stepped in the time axis direction. The target object data shifted by the shift amount when the calculated cross-correlation becomes maximum by calculating the cross-correlation between the target object data and the reference data in each shift amount. The electrophoretic measurement method according to claim 4, wherein a correlation coefficient between the reference data and the reference data is calculated. In the first electrophoresis step and the second electrophoresis step, detection data is acquired by sampling at a predetermined reference cycle,
At least one of the two expansion/contraction data and the measurement target data before expansion or the reference data acquired in the first expansion/contraction data acquisition step is resampled at a setting cycle different from the reference cycle. The electrophoretic measurement method according to claim 4 or 5. In the data correction step,
A second expansion/contraction data acquisition step of acquiring or expanding a plurality of expansion/contraction data based on the measurement target data in each expansion/contraction amount, in which the measurement target data is expanded or shortened stepwise in the time axis direction,
A second correlation coefficient calculating step of calculating a correlation coefficient between the plurality of expansion/contraction data and the reference data, and a correlation coefficient between the measurement target data before expansion/contraction and the reference data;
The expansion/contraction correction step of using the expansion/contraction data having the maximum correlation coefficient calculated by the second correlation coefficient calculation step or the measurement object data before expansion as the measurement object data after correction is included. The electrophoresis measurement method according to any one of claims 1 to 6. In the second correlation coefficient calculation step, the plurality of expansion/contraction data are stepwise shifted in the time axis direction, and the cross-correlation between the plurality of expansion/contraction data and the reference data in each shift amount is calculated, While calculating the correlation coefficient between the plurality of expansion/contraction data and the reference data shifted by the shift amount when the calculated cross-correlation becomes maximum, the measurement target data before expansion/contraction is stepped in the time axis direction. The target object data shifted by the shift amount when the calculated cross-correlation becomes maximum by calculating the cross-correlation between the target object data and the reference data in each shift amount. The electrophoretic measurement method according to claim 7, wherein a correlation coefficient between the reference data and the reference data is calculated. In the first electrophoretic step and the second electrophoretic step, detection data is acquired by sampling at a predetermined reference cycle,
At least one of the plurality of expansion/contraction data and the measurement target data before the expansion/contraction, or the reference data acquired in the second expansion/contraction data acquisition step is resampled at a setting cycle different from the reference cycle. The electrophoretic measurement method according to claim 7 or 8. In the data correction step,
A stretch correction step of stretching or shortening the measurement target data in the time axis direction,
The electrophoresis measurement method according to claim 1, further comprising a shift correction step of shifting the measurement target data in the time axis direction before or after the expansion/contraction correction step. In at least one of the reference data acquisition step and the measurement target data acquisition step, the reference intensity or the measurement target data is acquired by multiplying the detection intensity in each cycle of the detection data resampled at the set cycle by a coefficient. The electrophoretic measurement method according to any one of claims 1 to 10, wherein: In the first electrophoresis step, a plurality of reference samples containing different reference substances are electrophoresed to obtain a plurality of detection data corresponding to each reference sample,
In the reference data acquisition step, a plurality of reference data is acquired from the plurality of detection data acquired in the first electrophoresis step,
In the data correction step, with respect to the plurality of reference data, the measurement target data is relatively expanded or contracted or shifted in the time axis direction,
A similarity determination step of determining the similarity between the reference data and the measurement target data that is relatively expanded or contracted or shifted,
12. A matching data selecting step of selecting matching data from the plurality of reference data based on a judgment result in the similarity judging step, further comprising: Electrophoretic measurement method. The said similarity determination process determines similarity based on the correlation coefficient of each reference|standard data and the said measurement object data, and the peak area of the said measurement object data. Electrophoretic measurement method. In the similarity determination step, the similarity is determined based on the correlation coefficient between each reference data and the measurement target data, and the area ratio of the peak area of the measurement target data to the peak area of each reference data. The electrophoresis measurement method according to claim 12, wherein: In the similarity determination step, based on the correlation coefficient between each reference data and the measurement target data, the peak area of the measurement target data, and the area ratio of the peak area of the measurement target data to the peak area of each reference data. 13. The electrophoretic measurement method according to claim 12, wherein the similarity is determined. An average value of correlation coefficients calculated respectively in a plurality of regions included in the certain time width is used as a correlation coefficient between each reference data and the measurement target data. The electrophoretic measurement method according to any one of 1. A reference substance, a lower limit marker substance containing a component having an electrophoretic rate faster than the component contained in the reference substance, and an upper limit marker substance containing a component having a lower electrophoretic rate than the component contained in the reference substance were mixed. Detection data obtained by subjecting a reference sample to electrophoresis was obtained by normalizing to a certain time width with reference to each peak of the lower limit marker substance and the upper limit marker substance contained in the detection data. Standard data,
Measurement target substance, the lower limit marker substance containing a component having an electrophoretic velocity faster than the component contained in the measurement target substance, and the upper limit marker containing a component having an electrophoretic velocity slower than the component contained in the measurement target substance The detection data obtained by electrophoresing the sample to be measured in which the substance is mixed, with reference to the respective peaks of the lower limit marker substance and the upper limit marker substance contained in the detection data, in the constant time width A data processing device that performs data processing using measurement target data acquired by normalization,
A data processing device comprising: a data correction unit that relatively expands or contracts or shifts the reference data and the measurement target data in a time axis direction. A reference substance, a lower limit marker substance containing a component having an electrophoretic rate faster than that contained in the reference substance, and an upper limit marker substance containing a component having an electrophoretic rate slower than the component contained in the reference substance were mixed. Detection data obtained by subjecting a reference sample to electrophoresis was obtained by normalizing to a certain time width with reference to each peak of the lower limit marker substance and the upper limit marker substance contained in the detection data. Reference data,
Measurement target substance, the lower limit marker substance containing a component having an electrophoretic velocity faster than the component contained in the measurement target substance, and the upper limit marker containing a component having an electrophoretic velocity slower than the component contained in the measurement target substance The detection data obtained by electrophoresing the sample to be measured in which the substance is mixed, with reference to the respective peaks of the lower limit marker substance and the upper limit marker substance contained in the detection data, in the constant time width A data processing program for performing data processing using the measurement target data acquired by normalization,
A data processing program that causes a computer to function as a data correction unit that relatively expands or contracts the reference data and the measurement target data in the time axis direction.

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