JP5708537B2 - Electrophoresis device, electrophoresis method, and program for electrophoresis device - Google Patents

Description

  The present invention provides an electrophoresis apparatus for identifying a fragment contained in each sample by intermittently introducing a plurality of sample solutions into the flow path and detecting light emission of each sample solution passing through the flow path. The present invention relates to an electrophoresis method and a program for an electrophoresis apparatus.

  When analyzing a sample using electrophoresis, for example, a migration medium (separation medium) is filled in a flow path such as a capillary, and the sample solution is introduced into the flow path (for example, see Patent Document 1 below). In Patent Document 1, an internal standard substance whose electrophoresis speed is higher than that of a fragment contained in a sample is mixed with the sample solution, and the fragment contained in the sample is identified based on the detected peak of the internal standard substance. (Paragraphs [0006] and [0008] etc.)

  In the electrophoresis method as exemplified in Patent Document 1, the electrophoresis medium in the flow path is generally replaced every time each sample solution is analyzed. However, since it takes time to fill the electrophoresis medium in the flow path, when it is desired to analyze a plurality of sample solutions in a short time, the electrophoresis medium in the flow path is replaced every time each sample solution is analyzed. That is not preferable. Therefore, for example, a method of shortening the analysis time by introducing a plurality of sample solutions intermittently into the channel without replacing the electrophoresis medium in the channel until the electrophoresis medium deteriorates may be employed. is there.

  In addition, the internal standard substance in patent document 1 is used in order to analyze the sample solution with which the said internal standard substance is mixed. That is, Patent Document 1 does not disclose an electrophoresis method in which a plurality of sample solutions are intermittently introduced into a flow path, and the relationship between a plurality of sample solutions introduced intermittently is determined as an internal standard. It is not something that can be specified by substance.

JP 2007-24610 A

  In the electrophoresis method in which a plurality of sample solutions are intermittently introduced into the flow path, the analysis time can be further shortened by shortening the interval at which each sample solution is introduced. However, if the interval is too short, fragments contained in samples in different sample solutions cannot be distinguished from each other, and it may not be possible to recognize which sample contains the detected fragment.

  FIG. 8 is a diagram illustrating an example of a signal pattern detected when a plurality of sample solutions 1 are intermittently introduced into the flow path. In FIG. 8, for a plurality of sample solutions 1 introduced intermittently into the flow path, fragments F included in the samples in each sample solution 1 are shown in time series. In this example, a case where the sample solution 1a and the sample solution 1b are introduced into the flow path in this order will be described.

  As shown in FIG. 8 (a), after all the fragments Fa contained in the sample in the sample solution 1a previously introduced into the channel are detected, the sample solution 1b introduced into the channel later is detected. When the fragment Fb included in the sample is detected, it is possible to recognize which sample the detected fragments Fa and Fb are included in. However, if the interval at which the sample solutions 1a and 1b are introduced is too short, a part of the fragment Fb included in the sample in the sample solution 1b introduced into the flow path later as shown in FIG. 8B. May be detected before a part of the fragment Fa included in the sample in the sample solution 1a previously introduced into the flow path. In such a case, the fragments Fa and Fb included in the samples in the different sample solutions 1a and 1b cannot be distinguished from each other, and it is impossible to recognize which sample the detected fragment is included in. When such an error occurs, the reliability of the analysis result is lowered. However, conventionally, there has been no method capable of detecting the error from such a viewpoint.

  That is, even if it is easy to shorten the interval for introducing each sample solution, it has not been possible to determine whether or not an error has occurred. Whether or not an error has occurred is important information related to the reliability of the analysis results. Conventionally, the interval between introduction of each sample solution is generally sufficiently large so that no error always occurs. Met.

  The present invention has been made in view of the above circumstances, and when a plurality of sample solutions are intermittently introduced into a flow path, an error can be reliably detected even if the interval for introducing each sample solution is shortened. It is an object to provide an electrophoresis apparatus, an electrophoresis method, and a program for the electrophoresis apparatus that can be performed.

  The electrophoresis device according to the present invention intermittently introduces a plurality of sample solutions into a flow path, and identifies the fragments contained in each sample by detecting the light emission of each sample solution passing through the flow path. Luminescence of each sample solution mixed with a start marker whose electrophoresis speed is faster than the fragment contained in each sample and an end marker whose electrophoresis speed is slower than the fragment contained in each sample A detection unit for detecting the detection marker, a marker identification unit for identifying the start marker and the end marker mixed in each sample solution passing through the flow channel based on a detection result by the detection unit, and the flow channel Before the end marker mixed with any sample solution passing through the inside, the sample solution mixed with the sample solution introduced into the flow path after the sample solution If the start marker is identified by the marker identifying portion, characterized by comprising an error detection unit for detecting an error.

  According to such a configuration, when a plurality of sample solutions are intermittently introduced into the flow path, even if the interval between the introduction of each sample solution is shortened, the start marker and the end marker mixed in each sample solution By identifying the marker, an error can be detected reliably. That is, the start marker mixed with the sample solution introduced into the channel after the sample solution is identified before the end marker mixed with any sample solution passing through the channel. In such a case, there is a high possibility that fragments contained in those samples cannot be distinguished from each other. In such a case, it is possible to determine that the reliability of the analysis result is low by detecting an error in the error detection unit. Therefore, by using the error detection result by the error detection unit, it is possible to determine that the reliability of the analysis result is high when no error is detected.

  Moreover, the correction process regarding the fragment contained in each sample can also be performed using the start marker and the end marker. For example, the electrophoresis time of the fragment contained in each sample can be corrected with reference to the electrophoresis time of the start marker and the end marker. Therefore, by identifying the start marker and the end marker mixed in each sample solution, the result can be used for analysis.

  In addition, “luminescence” in the present invention includes not only “self-luminescence” in which a component contained in a sample solution itself emits light but also “fluorescence” in which light is emitted when a component contained in the sample solution is excited.

  At least one of the start marker and the end marker mixed in each sample solution may emit light at a wavelength different from the fragment contained in each sample. In this case, the detection unit may be able to identify and detect a plurality of wavelengths. According to such a configuration, in any case of analyzing any sample, based on the difference in the wavelength of light detected by the detection unit, the fragment included in each sample is changed to at least one of the start marker and the end marker. Can be identified.

  When both the start marker and the end marker emit light at the same wavelength as the fragment included in each sample, the fragment included in each sample is converted into the start marker and the fragment based on only the intensity of the signal obtained by the detection unit. Must be identified as an end marker. However, in this case, depending on the type of sample, there is a possibility that the intensity of the signal obtained by the detection unit is comparable between the fragment included in each sample and the start marker or the end marker. In such a case, it becomes difficult to distinguish the fragment included in each sample from the start marker and the end marker based only on the intensity of the signal obtained by the detection unit. Therefore, by adopting a configuration as in the present invention, it is possible to distinguish a fragment contained in each sample from at least one of a start marker and an end marker, even when analyzing any sample.

  The sample and at least one of the start marker and the end marker may have dyes that emit light at different wavelengths. In this case, the start marker and the end marker may be dye labels having predetermined wavelength characteristics.

  The fragment contained in each sample, the start marker, and the end marker may emit light at different wavelengths. In this case, the marker identification unit may identify the start marker and the end marker based on a difference in wavelength of light detected by the detection unit.

  According to such a configuration, since the fragment, the start marker, and the end marker all emit light at different wavelengths, they can be reliably identified. Thereby, since the possibility that an error is erroneously detected is reduced, the error can be detected more reliably.

  The start marker and the end marker mixed in each sample solution may emit light at the same wavelength. In this case, the marker identification unit may identify the start marker and the end marker based on a difference in signal strength obtained by the detection unit.

  According to such a configuration, by using the start marker and the end marker that emit light at the same wavelength, the types of wavelengths detected by the detection unit can be reduced, and thus the configuration of the apparatus can be simplified. is there. Thus, even when a start marker and an end marker that emit light at the same wavelength are used, the start marker and the end marker can be reliably identified based on the difference in the intensity of the signal obtained by the detection unit. it can. Even in the configuration of the present invention, since the start marker and the end marker emit light at a wavelength different from that of the fragment included in each sample, the start marker and the end marker are combined with the fragment included in each sample. Can be reliably identified.

  One of the start marker and the end marker mixed in each sample solution emits light at a wavelength different from the fragment included in each sample, and the other marker is the same as the fragment included in each sample. It may emit light at a wavelength. In this case, the marker identification unit may identify the one marker based on a difference in wavelength of light detected by the detection unit. Further, the error detection unit may detect an error based on the one marker identified by the marker identification unit and the number of fragments included in each sample.

  According to such a configuration, since the marker that emits light at the same wavelength as the fragment included in each sample can be used as the other marker, the types of wavelengths detected by the detection unit can be reduced. The configuration can be simplified. Even in the configuration of the present invention, the one marker emits light at a wavelength different from that of the fragment contained in each sample, so that the one marker and the fragment contained in each sample are reliably identified. can do. Therefore, an error can be reliably detected based on the one marker and the number of fragments included in each sample. Further, since the number of fragments contained in each sample is known, the other marker can be reliably identified.

  The electrophoresis method according to the present invention identifies a fragment contained in each sample by intermittently introducing a plurality of sample solutions into a channel and detecting light emission of each sample solution passing through the channel. Luminescence of each sample solution in which an initiation marker having a faster electrophoresis speed than the fragment contained in each sample and an end marker having a slower electrophoresis speed than the fragment contained in each sample are mixed A detection step for detecting the start marker, a marker identification step for identifying the end marker and the end marker mixed in each sample solution passing through the flow path based on the detection result in the detection step, and the flow Prior to the end marker mixed with any sample solution passing through the channel, mixed with the sample solution introduced into the flow path after the sample solution. When the start marker has been identified by the marker identifying step, characterized in that it comprises an error detecting step of detecting an error.

  The program for an electrophoresis apparatus according to the present invention includes a fragment included in each sample by intermittently introducing a plurality of sample solutions into the channel and detecting light emission of each sample solution passing through the channel. Is a program for an electrophoresis apparatus for identifying a start marker having a faster electrophoresis speed than a fragment contained in each sample and an end marker having a slower electrophoresis speed than a fragment contained in each sample. Detection step for detecting light emission of the sample solution, and marker identification step for identifying the start marker and the end marker mixed in each sample solution passing through the flow path based on the detection result in the detection step And before the end marker mixed in an arbitrary sample solution passing through the channel, and after the sample solution When the start marker is mixed in the introduced sample solution is identified by the marker identifying step, characterized in that to perform an error detecting step of detecting an error in the computer.

  According to the present invention, when a plurality of sample solutions are intermittently introduced into the flow path, the start marker and the end marker mixed in each sample solution can be reduced even if the interval for introducing each sample solution is shortened. By identifying, an error can be reliably detected.

It is a figure for demonstrating the outline of the electrophoresis method which concerns on one Embodiment of this invention. It is a block diagram which shows the structural example of the electrophoresis apparatus which concerns on this embodiment. It is a figure for demonstrating the specific aspect of the error detection by an error detection part. It is the flowchart which showed an example of the process by a control part. It is a figure for demonstrating the specific aspect of the error detection of the electrophoresis apparatus which concerns on another embodiment. It is a figure for demonstrating the specific aspect of the error detection of the electrophoresis apparatus which concerns on another embodiment. It is the flowchart which showed an example of the process by the control part in embodiment of FIG. It is the figure which showed an example of the signal pattern detected when a some sample solution is intermittently introduce | transduced in a flow path.

  FIG. 1 is a diagram for explaining an outline of an electrophoresis method according to an embodiment of the present invention. In the present embodiment, an example in which a plurality of sample solutions 1 are intermittently introduced into the capillary 2 will be described. The capillary 2 constitutes a flow path for the sample solution 1, and the capillary 2 is filled with an electrophoresis medium (separation medium) made of, for example, a gel. In the following description, a single capillary 2 will be described, but the same processing can be simultaneously performed on a plurality of capillaries 2.

  When analyzing a plurality of sample solutions 1 using electrophoresis, first, the capillary 2 is filled with a buffer solution (buffer), as shown in FIG. It is inserted into the first sample solution 1a. In this state, by applying a voltage between the cathode 3 inserted in the sample solution 1a together with the capillary 2 and an anode (not shown) provided on the other end side of the capillary 2, the sample solution 1a is introduced into the capillary 2 (injection).

  Thereafter, one end of the capillary 2 is inserted into the buffer solution 4 as shown in FIG. In this state, the sample solution 1 in the capillary 2 is applied by applying a voltage between the cathode 3 inserted in the buffer 4 together with the capillary 2 and the anode provided on the other end side of the capillary 2. Electrophoresis. During electrophoresis, the sample solution 1 is gradually separated (separated) while moving in the capillary 2.

  After electrophoresis for a predetermined time, as shown in FIG. 1C, one end of the capillary 2 is inserted into the second sample solution 1b. In this state, by applying a voltage between the cathode 3 inserted into the sample solution 1 b together with the capillary 2 and the anode provided on the other end side of the capillary 2, the sample solution 1 b is contained in the capillary 2. To be introduced.

  Again, as shown in FIG. 1 (d), one end of the capillary 2 is inserted into the buffer solution 4. In this state, the sample solution 1 in the capillary 2 is applied by applying a voltage between the cathode 3 inserted in the buffer 4 together with the capillary 2 and the anode provided on the other end side of the capillary 2. Electrophoresis.

  A plurality of sample solutions 1 can be intermittently introduced into the capillary 2 by repeating the above operation. At this time, by detecting the light emission of each sample solution 1 passing through the capillary 2, the fragments contained in each sample in each sample solution 1 can be identified. The component contained in each sample solution 1 may be self-luminous, or may be excited by excitation light to emit light (emit fluorescence).

  FIG. 2 is a block diagram illustrating a configuration example of the electrophoresis apparatus according to the present embodiment. The electrophoresis apparatus in this example includes a storage unit 20, an operation unit 30, a display unit 40, a detection unit 50, and the like in addition to the control unit 10 for controlling the operation of the apparatus.

  The control unit 10 includes, for example, a CPU (Central Processing Unit). When the CPU executes a program, the detection signal processing unit 11, the analysis processing unit 12, the error detection unit 13, the drive processing unit 14, and a display are displayed. It functions as the processing unit 15 and the like. The storage unit 20 can be configured by, for example, a ROM (Read-Only Memory) and a RAM (Random-Access Memory).

  The operation unit 30 is for a user to give instructions regarding the operation of the electrophoresis apparatus, and may include a keyboard or a mouse, for example. The display unit 40 is for displaying the operation status and analysis result of the electrophoretic device, and can be composed of, for example, a liquid crystal display.

  The detection unit 50 detects the light emission of each sample solution 1 passing through the capillary 2 and outputs a signal corresponding to the amount of received light. For example, when a component contained in each sample solution 1 emits light, the detection unit 50 includes at least a light receiving unit that receives light from each sample solution 1. On the other hand, when the component contained in each sample solution 1 emits fluorescence, the detection unit 50 includes at least a light emitting unit that irradiates each sample solution 1 with excitation light, and a light receiving unit that receives light from each sample solution 1. Is included.

  In this embodiment, each sample solution 1 is mixed with a start marker and an end marker that can be detected by the detection unit 50. The start marker is set so that the electrophoresis speed is faster than the fragment included in each sample. When the sample solution 1 is detected by the detection unit 50, the start marker is detected first in each sample solution 1. The start of each sample solution 1 can be identified. On the other hand, the end marker is set so that the electrophoresis speed is slower than the fragment included in each sample, and when the sample solution 1 is detected by the detection unit 50, the end marker is detected last in each sample solution 1. Thus, the end of each sample solution 1 can be identified.

  The start marker and the end marker mixed in each sample solution 1 are made of a dye label having a predetermined wavelength characteristic, for example. On the other hand, the sample may be one to which a dye is bound by intercalation. In this case, the dye as an intercalator that binds to the sample may be mixed in each sample solution 1 or may be mixed in the electrophoresis medium in the capillary 2.

  The detection signal processing unit 11 performs processing on an input signal from the detection unit 50. In this example, the detection signal processing unit 11 includes a marker identifying unit 111 and a fragment identifying unit 112. The marker identifying unit 111 identifies the start marker and the end marker mixed in each sample solution 1 passing through the capillary 2 based on the input signal from the detection unit 50. On the other hand, the fragment identification unit 112 identifies a fragment included in each sample in each sample solution 1 passing through the capillary 2 based on an input signal from the detection unit 50.

  The analysis processing unit 12 analyzes the fragments included in each sample identified by the fragment identifying unit 112. Through the processing of the analysis processing unit 12, fragments included in each sample are identified, and the analysis result is stored in the storage unit 20. In this example, a series of analysis results for a plurality of sample solutions 1 intermittently introduced into the capillary 2 are stored in the storage unit 20 for each series of analysis results.

  The error detection unit 13 detects an error based on the identification result by the marker identification unit 111. Specifically, for a plurality of sample solutions 1 that are intermittently introduced into the capillary 2, it becomes impossible to distinguish fragments contained in samples in different sample solutions 1, and the detected fragments are included in any sample. An error is detected when there is a risk that it may become impossible to recognize whether the fragment is a fragment. The error information detected by the error detection unit 13 is important information representing the reliability of a series of analysis results for a plurality of sample solutions 1 and is stored in the storage unit 20 in association with each analysis result. A specific mode of error detection by the error detection unit 13 will be described later.

  The drive processing unit 14 performs processing for controlling driving of each unit (for example, the detection unit 50) provided in the electrophoresis apparatus based on an input signal from the operation unit 30 or the like. The display processing unit 15 performs processing for controlling display on the display unit 40. In the present embodiment, each analysis result stored in the storage unit 20 can be displayed on the display unit 40 in association with error information. Therefore, the user can confirm not only the contents of each analysis result but also the reliability of each analysis result by confirming each analysis result and error information displayed on the display unit 40.

  FIG. 3 is a diagram for explaining a specific mode of error detection by the error detection unit 13. In FIG. 3, a plurality of sample solutions 1 that are intermittently introduced into the capillary 2 are included in the sample identified by the marker identifying unit 111 and the sample identified by the fragment identifying unit 112. Fragment F is shown in chronological order.

  In the present embodiment, before the end marker M2 mixed with an arbitrary sample solution 1 passing through the capillary 2, it is mixed with the sample solution 1 introduced into the capillary 2 after the sample solution 1. An error is detected when the start marker M1 is identified.

  For example, the case where the sample solution 1a and the sample solution 1b are introduced into the capillary 2 in this order will be described with reference to FIG. The sample solution 1a includes a fragment Fa, a start marker M1a, and an end marker M2a, and the sample solution 1b includes a fragment Fb, a start marker M1b, and an end marker M2b. As shown in FIG. 3A, after the end marker M2a mixed with the sample solution 1a previously introduced into the capillary 2 is identified, it is mixed with the sample solution 1b introduced into the capillary 2 later. If a start marker M1b is identified, no error is detected. On the other hand, as shown in FIG. 3 (b), after the start marker M1b mixed in the sample solution 1b introduced into the capillary 2 later is identified, the sample solution 1a introduced into the capillary 2 earlier is identified. If the mixed end marker M2a is identified, an error is detected.

  In the present embodiment, the fragment F, the start marker M1, and the end marker M2 included in each sample employ light emitting at different wavelengths. In order to make it easier to understand, in FIG. 3, different hatching is applied to the signal patterns of the fragment F, the start marker M1, and the end marker M2 included in each sample. In this case, the marker identifying unit 111 can identify the start marker M1 and the end marker M2 based on the difference in the wavelength of the light detected by the detection unit 50.

  FIG. 4 is a flowchart showing an example of processing by the control unit 10. During a series of analyzes performed by intermittently introducing a plurality of sample solutions 1 into the capillary 2, the control unit 10 identifies the start marker M1 of each sample solution 1 unless an error is detected by the error detection unit 13. (Yes in step S101), the process of identifying the end marker M2 of the sample solution 1 (Yes in step S102) is repeated. Then, when the end marker M2 of the final sample solution 1 is identified without detecting an error (Yes in step S103), the process ends as it is.

  On the other hand, after the start marker M1 is identified for any sample solution 1 (Yes in step S101), the end marker M2 is not identified (No in step S102), and the start marker M1 is identified. (Yes in step S104), the error detection unit 13 detects an error (step S105). In this case, since it is recognized that the start marker M1 mixed in the sample solution 1 introduced into the capillary 2 later is identified in step S104, error information is associated with the series of analysis results. By storing in the storage unit 20, it is possible to determine that the reliability of the analysis result is low.

  In the present embodiment, when a plurality of sample solutions 1 are intermittently introduced into the capillary 2, even if the interval at which each sample solution 1 is introduced is shortened, the start marker M 1 mixed with each sample solution 1 and By identifying the end marker M2, an error can be reliably detected. That is, the start mixed with the sample solution 1 introduced into the capillary 2 after the sample solution 1 before the end marker M2 mixed with the arbitrary sample solution 1 passing through the capillary 2 When the marker M1 is identified, there is a high possibility that the fragments F included in those samples cannot be distinguished from each other. In such a case, it is possible to determine that the reliability of the analysis result is low by detecting an error with the error detection unit 13. Therefore, by using the error detection result by the error detection unit 13, it is possible to determine that the reliability of the analysis result is high when no error is detected.

  In addition, correction processing related to the fragment F included in each sample can be performed using the start marker M1 and the end marker M2. For example, the electrophoresis time of the fragment F contained in each sample can be corrected based on the electrophoresis time of the start marker M1 and the end marker M2. Therefore, by identifying the start marker M1 and the end marker M2 mixed in each sample solution 1, it is also possible to use the result for analysis.

  Furthermore, in this embodiment, since the fragment F, the start marker M1, and the end marker M2 all emit light at different wavelengths, they can be reliably identified. Thereby, since the possibility that an error is erroneously detected is reduced, the error can be detected more reliably.

  However, the fragment F, the start marker M1, and the end marker M2 are not limited to configurations that emit light at different wavelengths. For example, the fragment F and the start marker M1, or the fragment F and the end marker M2, or the start marker. The configuration may be such that M1 and the end marker M2 emit light at the same wavelength. That is, it is only necessary that at least one of the start marker M1 and the end marker M2 mixed in each sample solution 1 emits light with a wavelength different from that of the fragment F included in each sample. In this case, the detection unit 50 needs to be configured to be able to identify and detect a plurality of wavelengths.

  With the above-described configuration, regardless of the sample to be analyzed, the fragment F included in each sample is converted into the start marker M1 and the sample based on the difference in the wavelength of light detected by the detection unit 50. It can be distinguished from at least one of the end markers M2.

  When both the start marker M1 and the end marker M2 emit light at the same wavelength as the fragment F included in each sample, the fragment F included in each sample is based only on the intensity of the signal obtained by the detection unit 50. Must be distinguished from the start marker M1 and the end marker M2. However, in this case, depending on the type of the sample, there is a possibility that the intensity of the signal obtained by the detection unit 50 is comparable between the fragment F included in each sample and the start marker M1 or the end marker M2. In such a case, it becomes difficult to distinguish the fragment F included in each sample from the start marker M1 and the end marker M2 based only on the intensity of the signal obtained by the detection unit 50. Therefore, with the above-described configuration, the fragment F included in each sample can be distinguished from at least one of the start marker M1 and the end marker M2 in any case of analyzing any sample.

  In the following, the above-described configuration, that is, a configuration in which at least one of the start marker M1 and the end marker M2 mixed in each sample solution 1 emits light at a wavelength different from that of the fragment F included in each sample. Another embodiment will be described. However, the configuration is not limited to such a configuration, and it is also possible to employ a configuration in which the fragment F, the start marker M1, and the end marker M2 all emit light at the same wavelength.

  FIG. 5 is a diagram for explaining a specific mode of error detection of the electrophoresis apparatus according to another embodiment. In FIG. 5, a plurality of sample solutions 1 that are intermittently introduced into the capillary 2 are included in the sample identified by the marker identifying unit 111 and the sample identified by the fragment identifying unit 112. Fragment F is shown in chronological order.

  In the present embodiment, the start marker M1 and the end marker M2 mixed in each sample solution 1 emit light at the same wavelength. In order to make this easy to understand, the same hatching is applied to the signal patterns of the start marker M1 and the end marker M2 in FIG. In this case, the start marker M1 and the end marker M2 are set so that the intensity of the signal obtained by the detection unit 50 is different in order to be identified by the marker identification unit 111. In this example, the start marker M1 is set to have a smaller signal intensity than the end marker M2. However, the present invention is not limited to this configuration, and the start marker M1 is more than the end marker M2. You may set so that the intensity | strength of the signal obtained may become large.

  In such a configuration, the marker identifying unit 111 can identify the start marker M <b> 1 and the end marker M <b> 2 based on the difference in signal strength obtained by the detection unit 50. The start of mixing with the sample solution 1 introduced into the capillary 2 before the end marker M2 mixed with the arbitrary sample solution 1 passing through the capillary 2 and after the sample solution 1 An error is detected when the marker M1 is identified.

  For example, the case where the sample solution 1a and the sample solution 1b are introduced into the capillary 2 in this order will be described with reference to FIG. The sample solution 1a includes a fragment Fa, a start marker M1a, and an end marker M2a, and the sample solution 1b includes a fragment Fb, a start marker M1b, and an end marker M2b. As shown in FIG. 5A, after the end marker M2a mixed with the sample solution 1a previously introduced into the capillary 2 is identified, it is mixed with the sample solution 1b introduced into the capillary 2 later. If a start marker M1b is identified, no error is detected. On the other hand, as shown in FIGS. 5B and 5C, after the start marker M1b mixed with the sample solution 1b introduced into the capillary 2 later is identified, it is introduced into the capillary 2 first. If the end marker M2a mixed in the sample solution 1a is identified, an error is detected.

  In the example of FIG. 5B, the sample solution 1a previously introduced into the capillary 2 is mixed with the sample solution 1b introduced into the capillary 2 before the fragment Fa contained in the sample. This shows a case where the start marker M1b is identified. In the example of FIG. 5C, the sample solution 1a introduced into the capillary 2 is mixed with the sample solution 1b introduced into the capillary 2 later after the fragment Fa contained in the sample in the sample solution 1a. The case where the start marker M1b is identified is shown. In any case, an error can be detected by the same processing as the processing described in FIG.

  In the present embodiment, since the types of wavelengths detected by the detection unit 50 can be reduced by using the start marker M1 and the end marker M2 that emit light at the same wavelength, the configuration of the apparatus can be simplified. is there. Thus, even when the start marker M1 and the end marker M2 that emit light at the same wavelength are used, the start marker M1 and the end marker M2 are surely determined based on the difference in the intensity of the signal obtained by the detection unit 50. Can be identified. Even in the configuration of the present embodiment, since the start marker M1 and the end marker M2 emit light at a wavelength different from that of the fragment F included in each sample, the start marker M1, the end marker M2, and each sample Can be reliably identified.

  FIG. 6 is a diagram for explaining a specific mode of error detection of the electrophoresis apparatus according to still another embodiment. In FIG. 6, a plurality of sample solutions 1 intermittently introduced into the capillary 2 are included in the sample identified by the marker identifying unit 111 and the start marker M1 and end marker M2 identified by the fragment identifying unit 112. Fragment F is shown in chronological order.

  In the present embodiment, among the start marker M1 and the end marker M2 mixed in each sample solution 1, the start marker M1 emits light at a wavelength different from that of the fragment F included in each sample. On the other hand, the end marker M2 emits light at the same wavelength as the fragment F included in each sample. In order to make it easier to understand, in FIG. 6, the same hatching is applied to the signal patterns of the fragment F and the end marker M2, and the signal patterns of the start marker M1 are applied to different hatchings.

  In such a configuration, the marker identifying unit 111 can identify the start marker M1 based on the difference in the wavelength of the light detected by the detecting unit 50. In this embodiment, the error detection unit 13 detects an error based on the start marker M1 identified by the marker identification unit 111 and the number of fragments F included in each sample.

  That is, the marker identifying unit 111 can identify the end marker M2 detected after the fragment F by counting the number of fragments F detected after identifying the start marker M1. The start of mixing with the sample solution 1 introduced into the capillary 2 before the end marker M2 mixed with the arbitrary sample solution 1 passing through the capillary 2 and after the sample solution 1 When the marker M1 is identified, an error is detected.

  For example, the case where the sample solution 1a and the sample solution 1b are introduced into the capillary 2 in this order will be described with reference to FIG. The sample solution 1a includes a fragment Fa, a start marker M1a, and an end marker M2a, and the sample solution 1b includes a fragment Fb, a start marker M1b, and an end marker M2b. As shown in FIG. 6A, after the end marker M2a mixed with the sample solution 1a previously introduced into the capillary 2 is identified, it is mixed with the sample solution 1b introduced into the capillary 2 later. If a start marker M1b is identified, no error is detected. On the other hand, as shown in FIG. 6 (b), after the start marker M1b mixed with the sample solution 1b introduced into the capillary 2 later is identified, the sample solution 1a introduced into the capillary 2 earlier is identified. If the mixed end marker M2a is identified, an error is detected.

  FIG. 7 is a flowchart showing an example of processing by the control unit 10 in the embodiment of FIG. During a series of analyzes performed by intermittently introducing a plurality of sample solutions 1 into the capillary 2, the control unit 10 identifies the start marker M1 of each sample solution 1 unless an error is detected by the error detection unit 13. (Yes in step S201), the number of fragments F contained in the sample in the sample solution 1 is counted (step S202), and it is determined whether or not the number reaches a specified number (step S203). Repeat the process.

  The specified number is one larger than the number of fragments F included in each sample. That is, if the number of fragments F included in each sample is “3”, the specified number is “4”. Thereby, when the specified number is counted (Yes in step S203), the signal detected by the detection unit 50 at that time can be identified as the end marker M2. In the processing as described above, when the end marker M2 of the final sample solution 1 is identified without detecting an error (Yes in step S204), the processing ends as it is.

  On the other hand, for any sample solution 1, after the start marker M1 is identified (Yes in step S201), the specified number is not counted (No in step S203), and the start marker M1 is identified. (Yes in step S205), the error detector 13 detects an error (step S206). In this case, since it is recognized that the start marker M1 mixed in the sample solution 1 introduced into the capillary 2 later is identified in step S205, error information is associated with the series of analysis results. By storing in the storage unit 20, it is possible to determine that the reliability of the analysis result is low.

  In the present embodiment, since the marker that emits light at the same wavelength as the fragment F included in each sample is used as the end marker M2, the types of wavelengths detected by the detection unit 50 can be reduced. It can be simplified. Even in the configuration of the present embodiment, since the start marker M1 emits light at a wavelength different from that of the fragment F included in each sample, the start marker M1 and the fragment F included in each sample can be reliably detected. Can be identified. Therefore, an error can be reliably detected based on the start marker M1 and the number of fragments F included in each sample. In addition, since the number of fragments F included in each sample is known, the end marker M2 can also be reliably identified.

  However, the present invention is not limited to the above configuration. For example, among the start marker M1 and the end marker M2 mixed in each sample solution 1, the end marker M2 emits light at a wavelength different from the fragment F included in each sample. Such a configuration may be adopted. In this case, the start marker M1 may be configured to emit light at the same wavelength as the fragment F included in each sample.

  In the above embodiment, the detection step for detecting the luminescence of each sample solution 1, the marker identification step for identifying the start marker M1 and the end marker M2 based on the detection result in the detection step, and the identification result in the marker identification step In the above description, the error detection step for detecting an error based on the above is executed by the electrophoresis apparatus. In this case, a program for causing a computer to execute each step may be provided separately from the electrophoresis apparatus. Thereby, the computer which controls using the said program functions as an electrophoresis apparatus, and there can exist an effect similar to the said embodiment. It is also possible to provide the program stored in a storage medium.

  Further, the configuration is not limited to a configuration in which each of the above steps is executed by a program, and a configuration in which at least a part of the steps is executed by a human may be employed. In this case, the configuration may be such that only a part of each step is executed by a human, or the configuration may be such that all steps are executed by a human.

  Furthermore, the present invention can be applied not only to capillary electrophoresis in which the sample solution 1 is introduced into the capillary 2 but also to various other types of electrophoresis. For example, a configuration in which electrophoresis is performed by a so-called cross injection method may be employed. In this case, after loading the sample solution 1 into the first channel between the well and the drain, a voltage is applied to both ends of the second channel orthogonal to the first channel, thereby the second channel. A configuration in which the sample solution 1 is introduced intermittently may be employed. When the present invention is applied to the cross-injection method, for example, it is possible to employ a configuration in which a plurality of wells are provided and each well is connected to a branch path branched from the first flow path.

  The present invention can be applied to an analyzer for analyzing a biomolecule such as DNA, RNA, or protein. In this case, the present invention can also be applied to, for example, an analyzer used for quality control of food. However, the present invention is not limited to these and can be applied to all other technical fields.

  For example, when the present invention is applied to a DNA sequencer (DNA analyzer), the effects of the present invention can be achieved by simply applying the program of the present invention to an existing hardware configuration. That is, in an analyzer equipped with a detection unit capable of identifying and detecting a plurality of wavelengths, such as a DNA sequencer, the above-described program can be applied only by applying the program of the present invention without newly adding a hardware configuration. A configuration similar to that of the embodiment can be obtained. Thereby, the application range of various analyzers such as a DNA sequencer can be expanded.

DESCRIPTION OF SYMBOLS 1 Sample solution 2 Capillary 3 Cathode 4 Buffer 10 Control part 11 Detection signal processing part 12 Analysis processing part 13 Error detection part 14 Drive processing part 15 Display processing part 20 Storage part 30 Operation part 40 Display part 50 Detection part 111 Marker identification part 112 Fragment identification part M1 start marker M2 end marker

Claims (7)

An electrophoretic apparatus for intermittently introducing a plurality of sample solutions into a flow path and detecting fragments of each sample solution passing through the flow path to identify fragments contained in each sample,
A detection unit for detecting luminescence of each sample solution mixed with a start marker having a faster electrophoresis speed than the fragment contained in each sample and an end marker having a slower electrophoresis speed than the fragment contained in each sample;
Based on the detection result by the detection unit, a marker identification unit for identifying the start marker and the end marker mixed in each sample solution passing through the flow path,
The start marker mixed with the sample solution introduced into the flow path after the sample solution before the end marker mixed with any sample solution passing through the flow path is An electrophoresis apparatus, comprising: an error detection unit that detects an error when the marker is identified by the marker identification unit. At least one of the start marker and the end marker mixed in each sample solution emits light at a wavelength different from the fragment included in each sample,
The electrophoresis device according to claim 1, wherein the detection unit is capable of identifying and detecting a plurality of wavelengths. The fragment contained in each sample, the start marker and the end marker emit light at different wavelengths,
The electrophoresis apparatus according to claim 2, wherein the marker identifying unit identifies the start marker and the end marker based on a difference in wavelength of light detected by the detection unit. The start marker and the end marker mixed in each sample solution emit light at the same wavelength,
The electrophoresis apparatus according to claim 2, wherein the marker identification unit identifies the start marker and the end marker based on a difference in intensity of signals obtained by the detection unit. One of the start marker and the end marker mixed in each sample solution emits light at a wavelength different from the fragment included in each sample, and the other marker is the same as the fragment included in each sample. Which emits light at a wavelength,
The marker identifying unit identifies the one marker based on a difference in wavelength of light detected by the detecting unit,
The electrophoresis according to claim 2, wherein the error detection unit detects an error based on the one marker identified by the marker identification unit and the number of fragments included in each sample. apparatus. An electrophoresis method for identifying a fragment contained in each sample by intermittently introducing a plurality of sample solutions into a flow path and detecting light emission of each sample solution passing through the flow path,
A detection step for detecting luminescence of each sample solution mixed with a start marker having a faster electrophoresis speed than the fragment contained in each sample and an end marker having a slower electrophoresis speed than the fragment contained in each sample;
Based on the detection result in the detection step, a marker identification step for identifying the start marker and the end marker mixed in each sample solution passing through the flow path;
The start marker mixed with the sample solution introduced into the flow path after the sample solution is ahead of the end marker mixed with any sample solution passing through the flow path. And an error detection step of detecting an error when the marker is identified in the marker identification step. A program for an electrophoretic device that identifies a fragment contained in each sample by intermittently introducing a plurality of sample solutions into the flow channel and detecting light emission of each sample solution passing through the flow channel. ,
A detection step for detecting luminescence of each sample solution mixed with a start marker having a faster electrophoresis speed than the fragment contained in each sample and an end marker having a slower electrophoresis speed than the fragment contained in each sample;
Based on the detection result in the detection step, a marker identification step for identifying the start marker and the end marker mixed in each sample solution passing through the flow path;
The start marker mixed with the sample solution introduced into the flow path after the sample solution is ahead of the end marker mixed with any sample solution passing through the flow path. A computer program for an electrophoresis apparatus, which causes a computer to execute an error detection step of detecting an error when the marker is identified in the marker identification step.

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