JP6796561B2 - Biological sample analyzer and method - Google Patents

Description

The present invention relates to an analyzer that performs sequence analysis of nucleic acids using a thin film having nano-sized pores, and particularly relates to a technique for improving the accuracy of nucleotide sequence decoding with respect to environmental vibration.

As a background technique in this technical field, there is Patent Document 1 relating to nanopore DNA sequencing. In this publication, it is stated that "the inner diameter of the nanopore was about 10 nm." The nanopore DNA sequencer is a method of directly electrically measuring the base sequence of DNA without performing an extension reaction or a fluorescent label. Is in the spotlight. Several methods have been proposed for this direct measurement method, one of which is the blockade current method. Pore (nanopore) of several nm is prepared on the thin film by a transmission electron microscope or the like, and liquid tanks filled with an electrolyte solution are provided on both sides of the thin film. When electrodes are provided in each liquid tank and a voltage is applied between these electrodes, an ionic current flows through the nanopores. The ionic current is proportional to the cross-sectional area of the nanopore as a first-order approximation. When the DNA passes through the nanopores, the DNA blocks the nanopores, reducing the effective cross-sectional area through which the ions can pass, thus reducing the ion current. This amount of decrease is called the blocking current. Based on the magnitude of the blocking current, the difference between single-strand and double-strand DNA and the type of base are determined.

Since there are problems such as the large Brownian motion of DNA passing through the nanopores and the speed of passing through the nanopores being too fast for the detector to catch up, it is difficult to ensure the measurement accuracy for distinguishing each base. .. In order to control the Brownian motion and transit speed of DNA, several methods have been proposed in which one end of DNA is fixed and the motion is controlled before measurement. A method of fixing DNA to a cantilever used in an AFM (Atomic Force Microscope) or the like (Patent Document 2), a method of fixing a biological sample to a nanostepper arm mounted on a stage (Patent Document 3), and the like are disclosed.

Then, when fixing one end of a single strand or double strand of DNA to a plane parallel to the plane on which the nanopores are created and driving the DNA on an axis perpendicular to that plane, the nanopore passage speed of DNA is controlled. Further, there may be a case where a sample transfer stage is provided so that the position of the single or double strand of DNA can be positioned at a desired position on the nanopore.

International Publication No. WO2012-043028A1 U.S. Patent Publication No. 2004-014658 Japanese Unexamined Patent Publication No. 2006-078491

One drawback of the conventional DNA transfer technique described above is the vibration between the thin film in which the nanopores, which are the detectors, are opened and the driving component that controls the passing speed of the DNA. For a DNA sequencer that is required to accurately decode the base sequence, it is extremely difficult to control it ultra-precisely without reversing the sequence order of DNA whose base-to-base distance is sub-nanometer, such as floor vibration and acoustic vibration. Due to the environmental vibration of the DNA, vibration may occur between the thin film in which the nanopores that are the detectors are opened and the driving component that controls the passing speed of DNA. On the other hand, a method of physically suppressing environmental vibration by using an active vibration isolator or the like and not reversing the sequence order of DNA is also conceivable. As described above, in the nanopore DNA sequencer system, if the order of the base sequences is reversed due to environmental vibration, accurate sequence analysis cannot be performed.

According to the technique disclosed in Patent Document 2, one end of DNA is fixed to a cantilever and moved up and down. In general AFM, the amount of deflection of the cantilever is measured by some method and feedback control is performed. Since what can be measured is the amount of deflection of the cantilever, there is a problem that the vibration between the thin film in which the nanopore, which is a detector, is opened and the cantilever that controls the passing speed of DNA cannot be measured.

Further, according to the technique disclosed in Patent Document 3, one end of DNA is fixed to an electrostatic actuator called a nanostepper, generally a piezostage, to perform biaxial operation. The piezo stage measures the moving parts in some way and performs feedback control. Even with this configuration, what can be measured is the amount of movement of the piezo stage, so the vibration between the thin film with the nanopores that are the detectors opened and the nanostepper that controls the DNA passage speed is measured. There was a problem that I couldn't do.

As described above, in the conventional configuration, the vibration between the thin film in which the nanopores that are the detectors are opened and the driving component that controls the passage speed of DNA is not directly measured, and floor vibration or acoustic vibration is not used. When relative vibration occurs due to environmental vibration such as, the order of the base sequence of DNA is reversed, and there is a problem that accurate sequence analysis cannot be performed.

An object of the present invention is to provide a biological sample analyzer capable of solving the above-mentioned problems and solving a decrease in base sequence decoding accuracy due to environmental vibration, and an analysis method thereof.

In order to achieve the above object, in the present invention, it is a bioanalyzer that analyzes the base sequence of DNA, and the inside is separated by a nanopore substrate having a nanometer-sized hole through which a measurement object passes and a nanopore substrate. A tank filled with an electrolyte solution, a DNA-fixed substrate that is located facing the nanopore substrate and on which DNA is fixed, a drive mechanism that drives the DNA-fixed substrate, and a voltage that applies voltage to both sides of the nanopore substrate. It includes an application unit, a detection unit that detects an ion current when DNA passes through the nanopore substrate, a vibration detection unit that detects the vibration of the bioanalyzer, and a recording unit that records the vibration information detected by the vibration detection unit. A biological sample analyzer having a configuration is provided.

Further, in order to achieve the above object, the present invention is a bioanalytical method of a bioanalyzer that analyzes a base sequence of DNA, and the bioanalyzer is a nanometer-sized hole through which a measurement object passes. A tank filled with an electrolyte solution, a DNA-fixed substrate facing the nanopore substrate, and a DNA-fixed substrate, and a DNA-fixed substrate, whose inside is separated by a nanopore substrate to which a voltage is applied on both sides of the nanopore substrate. It is equipped with a drive mechanism that drives the device, a detection unit that detects the ion current when the DNA passes through the nanopore substrate, and a vibration detection unit that detects the vibration information of the bioanalytical device. Provided is a biological sample analysis method for analyzing a base sequence using the vibration information obtained.

According to the present invention, even if a vibration in which the base sequence order is reversed occurs, it can be detected and recorded, and the base sequence decoding can be improved by excluding or correcting the blocking signal at the time of occurrence of the vibration.

It is a figure which shows an example of the structure of the biological sample analyzer and the enlarged view of the biological sample analysis chip. It is a figure which shows an example of the sequence reading signal acquired by a biological sample analysis chip. It is a figure which shows an example of the case where the blockade current without vibration is measured by the biological sample analyzer. It is a figure which shows an example of the case where the blockade current with vibration is measured by the biological sample analyzer. It is a figure which shows one structural example of the biological sample analyzer and the analysis method which concerns on Example 1. FIG. It is a figure which shows an example of the vibration signal measured at the same time as the sequence reading example acquired by the biological sample analysis chip which concerns on Example 1. FIG. It is a figure which shows one structural example of the biological sample analyzer and method which concerns on Example 2. FIG. It is a figure which shows one structural example of the biological sample analyzer and method which concerns on Example 3. FIG. It is a figure which shows one structural example of the biological sample analyzer and method which concerns on Example 4. FIG. It is a figure which shows one structural example of the biological sample analysis method which concerns on Example 5. It is a figure which shows one structural example of the biological sample analyzer and method which concerns on Example 6.

Hereinafter, examples of the present invention will be described with reference to the drawings. Although the drawings show specific examples based on the principles of the present invention, these are for the purpose of understanding the present invention and are not used for the limited interpretation of the present invention. Deoxyribonucleic acid (DNA) is exemplified as a living body to be analyzed, but it is not limited to DNA and may be nucleic acid such as ribonucleic acid (RNA).

The “nanopore” described in each embodiment of the present specification is a nano-sized hole provided in the thin film and penetrates the front and back surfaces of the thin film. The thin film is mainly formed from an inorganic material. Further, the substrate or beads to which one end of the DNA fragment is fixed are mainly formed from an inorganic material. The material of the thin film, the substrate or the beads may also include an organic substance, a polymer material and the like.

First, a biological sample analyzer using a nanopore device according to Example 1 will be described. FIG. 1 is a diagram showing an example of the configuration of the biological sample analyzer of this embodiment, and shows a base sequence reading mechanism by a biological sample analysis chip.

As shown in FIG. 1, a biological sample analyzer 100 such as a DNA analyzer, for example, includes two tanks 102A and 102B separated by a partition 101. A biological sample analysis chip having nanopores (hereinafter referred to as a nanopore device) is installed in the partition body 101. The two tanks 102A and 102B are filled with the electrolyte solution 103. The two tanks 102A and 102B are electrically connected by an electrode 104 and a power supply 105A, which are voltage application portions for applying voltage to both sides of the nanopore device. The entire flow path partitioned by this nanopore device is called a flow cell 106.

As shown in the enlarged view in the circle 100A of FIG. 1, one end of the DNA is chemically fixed to the DNA fixing substrate 108 attached to the probe 107, and when the DNA is entered through the flow cell opening 110 by the drive mechanism 109, it is in the liquid. DNA 111 migrates from one tank 102A to the other tank 102B through the nanopore 112 of the nanopore device of the partition 101, but does not pass by. By pulling out or pushing one end of the DNA 111 introduced into the nanopore 112 by the drive mechanism 109, the current value is measured by the ammeter 105B, which is a detection unit that detects the ion current. Since the current value changes as the DNA 111 passes through the nanopore 112 of the nanopore device, the base sequence is read from this current value. The current value measured by the ammeter 105B is amplified by an amplifier (not shown) and recorded in a storage unit of a personal computer (PC) (not shown) via an A / C converter (not shown). ..

The nanopore device of the partition plate 101 includes a nanopore 112 and a nanopore thin film 113 having a nanopore of several nm, for example, 5 nm. The area of the very thin nanopore thin film 113 is small, and the thickness of the partition 101 on which the nanopore device is installed is several hundred nm, although not shown, for reinforcement. When the electrolyte solution 103 is filled in the upper and lower tanks 102A and 102B of the partition body 101 and a voltage is applied up and down through the nanopore device, a current derived from ions in the electrolyte solution 103 according to the cross-sectional area of the pore diameter of the nanopore 112. Is detected. When the DNA 111 passes through the nanopore 112 by the drive stage, the flow of ions is obstructed, so that the current value is reduced by the cross-sectional integral of the DNA 111 in the nanopore 112. Although the DNA 111 has been described using beads that are spheres in the description, the base has a shape based on a chemical formula. Hereinafter, thread-like (single-strand) or spherical (bead) -shaped DNA 111 will be described as appropriate.

The DNA analyzer 100 performs base identification based on the difference in the amount of change in the current value for each base type. FIG. 2 is a diagram showing an example of a current change for each base type that can be read by applying a voltage to the electrode 104 and pulling it out or pushing it in by the drive mechanism 109. As shown in the sequence reading example 200, different current values are detected for each of the four types of bases A, C, T, and G, and the base sequence is decoded by driving the DNA 111 in one direction.

Here, the details of the biological sample analyzer will be described with reference to FIG. FIG. 3A of the figure shows a biological sample analyzer 300 having the same configuration as the biological sample analyzer 100, and an enlarged view 301 of DNA 111 is shown on the right side thereof. The DNA fixing substrate 108 to which the DNA 111 is attached is moved in one direction by the drive mechanism 109. Here, assuming that the ion current value moves in the + Z direction, data such as the current waveform 302 shown in (b) of the figure is recorded as the ion current value measured by the ammeter 105B. In order to obtain accurate sequence information, it is important to move in one direction without reversing the sequence order.

Further, the enlarged view 301 of the DNA 111 is a schematic diagram, and it is possible to directly measure the place where the DNA 111, which has a sub-nanometer distance between bases of 0.34 nm, passes through the nanopore 112 having a nanopore diameter of 1 to 2 nm. It is very difficult due to the resolution, and it is difficult to observe a biological sample containing a liquid with an electron microscope in which the observation sample must be evacuated. There are also atmospheric pressure electron microscopes, but this is also a problem of resolution, and high-precision observation is difficult.

The operation of the biological sample analyzer 300 when the actual operating state is taken into consideration will be described with reference to FIG. As shown in (a) of the figure, the environment in which the biological sample analyzer 300 is installed includes environmental vibrations such as floor vibration 401 and acoustic vibration 402, and the entire biological sample analyzer 300 vibrates. Since the distance between the bases of the DNA 111 is sub-nanometer, the biological sample analyzer 300 vibrates even with a very small vibration, and the thin film 113 in which the nanopores that are the detectors are opened and the drive mechanism 109 via the probe 107 Vibration is generated between the attached DNA fixing substrate 108 and the sequence order is reversed. As the current value measured by the ammeter 105B when the arrangement order is reversed, data such as the current waveform 403 in (b) of the figure is recorded. That is, it is the value of the blocking current when the vibration that reverses the sequence order is included in the time of the alternate long and short dash line of the current waveform 403, that is, when the DNA reversion occurs. Due to environmental vibration, the sequence information obtained from the current waveform 403 is (ACTGA GA CTGA) with respect to the original DNA 111 in the base sequence order (ACTGACTGA) shown in the enlarged view 301 of (a) of the same figure, and is underlined. Since the DNA is reversed at the subtracted GA portion, that is, the sequence order is reversed, accurate sequence information cannot be obtained, which greatly affects the decrease in base sequence decoding accuracy.

FIG. 5 shows an analysis execution device and a method of the biological sample analyzer 500 of this example. An enlarged view 301 of the nanopore device of the partition body 101 is shown in a circle. It is configured to include at least one accelerometer 501, an A / C converter 502, and a PC 503, which are vibration detection units for detecting the vibration of the analyzer, such as in the biological sample analyzer 500 or on the floor on which the analyzer is installed. The current value measured by the ammeter 105B and the signal value measured by the accelerometer 501 are recorded in synchronization with the PC 503 which is a recording means via the A / C converter 502. The vibration detection unit may be, for example, a speedometer or a displacement meter.

FIG. 6 shows the biological sample analyzer 500 of this embodiment via the A / C converter 502 when simultaneously measuring the blocking current (Ion Current) and the vibration signal (Acceleration) when the vibration is generated due to the environmental vibration. The waveform 600 recorded in PC503 is shown. The vibration generated between the thin film 113 in which the nanopores, which are detectors, are opened by the environmental vibration and the DNA fixing substrate 108 attached to the drive mechanism 109 via the probe 107 affects the amount of blocking current. It is difficult to accurately sequence the original DNA 111 base sequence sequence (ACTGACTGA) using only the blocking current.

However, in the biological sample analyzer 500 of this embodiment, by simultaneously measuring the blocking current and the vibration signal, it is detected and recorded that vibration is generated during the period during which the vibration signal is detected. Then, the accuracy can be improved by excluding the blocking current value at the time of vibration generation without using it for base sequence decoding by the alarm unit 504 that controls, for example, displaying an alarm on the display of the PC 503. .. The alarm unit 504 can be realized by creating and executing a program of the PC 503.

According to the configuration of this embodiment, by simultaneously measuring not only the blocking current but also the device vibration due to the environmental vibration such as the floor vibration and the acoustic vibration, the vibration due to the environmental vibration is not controlled to the nanometer order or less. Even if vibration in which the sequence order of DNA is reversed occurs, it is detected and recorded, and the blocking signal at the time of vibration is excluded to improve the decoding of the base sequence. Therefore, there is an effect that the base sequence decoding accuracy can be improved, the number of devices that physically suppress vibration to sub-nanometers, for example, expensive devices such as an active vibration isolator can be reduced, and the environmental vibration specifications can be greatly expanded.

FIG. 7 shows a configuration example of the biological sample analyzer of Example 2. Similar to the biological sample analyzer 500 of FIG. 5, at least one accelerometer 501, which is a vibration detection unit for detecting the vibration of the analyzer, is provided in the biological sample analyzer 700 or on the floor where the biological sample analyzer is installed. Is. The current value measured by the ammeter 105B and the signal value measured by the accelerometer 501 are recorded in synchronization with the PC503 which is a recording means via the A / C converter 502, and further, in this embodiment, it is necessary. Correspondingly, a command is given to the drive mechanism 109 by executing the program of the alarm unit 504 of the PC 503. Further, as in the first embodiment, the vibration detection unit may be, for example, a speedometer or a displacement meter.

In the configuration of this embodiment, when vibration is generated in the biological sample analyzer 700 due to environmental vibration, the blocking current and the vibration signal are simultaneously measured, and when the waveform 600 as shown in FIG. 6 is measured, an alarm is issued. The alarm unit 504 that emits a command to stop the drive of the drive mechanism 109 to which the DNA fixing substrate 108 is attached via the probe 107 when vibration is generated, and after the vibration has subsided, the drive is restarted to achieve accuracy. It is possible to improve. It is also possible to move to the position of the base sequence when it starts to vibrate by the drive mechanism 109, measure the ion current with the ammeter 105B, and remeasure the blocking current.

FIG. 8 shows a configuration example of the biological sample analyzer of Example 3. This example is an example of the biological sample analyzer 800 having a configuration in which the active vibration isolator 801 is further provided in the biological sample analyzer 700 of the second embodiment. The active vibration isolation table 801 includes a sensor 805 which is a vibration detection unit that detects vibration of at least one or more analyzers, and at least one or more springs 803A, B and an actuator 804A for insulation from the installed floor. , B, and a plate or surface plate 802 on which a nanopore sequencer is installed. By placing the biological sample analyzer on the active vibration isolation table, the influence of floor vibration and the like can be reduced. Similar to the second embodiment, the current value measured by the ammeter 105B and the signal value measured by the sensor 805 are recorded in synchronization with the PC503, which is a recording means, via the A / C converter 502, and if necessary, It is assumed that a command is given to the drive mechanism 109 from the PC 503. Further, the vibration detection unit may be, for example, a speedometer or a displacement meter.

When the biological sample analyzer 800 of this embodiment vibrates due to environmental vibration, especially floor vibration, the blocking current and the vibration signal are simultaneously measured, and when the waveform 600 as shown in FIG. 6 is measured, an alarm is issued. By the processing of the alarm unit 504 that is issued, when vibration is generated, the drive mechanism 109 to which the DNA fixing substrate 108 is attached is instructed to stop via the probe 107, and after the vibration has subsided, it is driven again to improve the accuracy. Is possible. There is also a method in which the drive mechanism 109 moves to the position of the base sequence when the vibration starts and the blocking current is remeasured.

FIG. 9 shows a configuration example of the biological sample analyzer and method of Example 4. In the biological sample analyzer 900 of this embodiment, the displacement meter 902 that eliminates the DNA and the electrolyte solution and measures the DNA fixing substrate 108 based on the nanopore substrate, and the vibration detector that detects the vibration of at least one or more analyzers. As a configuration including the sensor 901, the environmental vibration is measured and the transfer function 903 from the sensor 901 to the displacement meter 902 is acquired in advance. Again, the vibration detection unit may be, for example, a speedometer or a displacement meter.

The transfer function 903 acquired in advance with the above configuration Laplace transforms the value of the sensor 901 that measured the environmental vibration into X (s), and Laplaces the value of the displacement meter 902 that measured the DNA fixing substrate 108 based on the nanopore substrate. It is known that the transformed Y (s) and G (s) are calculated as in Equation 1.

When the biological sample analyzer 700 shown in FIG. 7 vibrates due to environmental vibration, the partition body 101 in which the nanopore, which is a detector, is opened from the device vibration by the transfer function 903 acquired in advance by the above equation. , It is possible to predict the vibration between the DNA fixing substrate 108.

Then, when the blocking current and the vibration signal are simultaneously measured and the waveform 600 as shown in FIG. 6 is measured, the partition body 101 in which the nanopores, which are detectors, are opened from the vibration signal by the transfer function 903, and the DNA fixing substrate 108. The alarm unit 504, which predicts the vibration between the two and issues an alarm, can perform feedback control to the drive mechanism 109 to which the DNA fixing substrate 108 is attached via the probe 107. That is, when the vibration detection unit detects the vibration, the alarm unit 504 feeds back the vibration information to the drive mechanism by the transfer function and drives the DNA fixing substrate 108 so as to suppress the vibration of the DNA. With the configuration of this embodiment, it is possible to suppress the vibration between the partition body in which the nanopore is opened and the DNA fixing substrate, and to improve the accuracy of base sequence decoding.

FIG. 10 is a diagram for explaining the biological sample analysis method of Example 5. As shown in the current waveform 1000 in the figure, the blockage current and the vibration signal are measured at the same time, and the partition body 101 in which the nanopore, which is a detector, is opened from the vibration signal by the transfer function 903, and the DNA fixing substrate 108. The merit of predicting displacement vibration is described. When the transfer function 903 is acquired in the configuration of the fourth embodiment, accurate prediction can be performed by using the high-precision displacement meter 902. From accurate displacement vibration prediction, the behavior of DNA in units of one base can be predicted, and even if the sequence order is reversed, accurate sequence information can be obtained by correction using a vibration signal. That is, there is a merit that accurate sequence information can be obtained without physically controlling the nanometer order.

The lower part of FIG. 10 shows an enlarged view 1001 of the nanopore device. The environment in which the biological sample analyzer 500 is installed as shown in FIG. 5 includes floor vibration 401 and acoustic vibration 402, and the current value measured by the ammeter 105B when the entire biological sample analyzer 500 vibrates is , Data such as current waveform 1000 is recorded. The current waveform 1000 shows the value of the blocking current when a vibration that reverses the arrangement order is input at the time of the dotted line. In the example of FIG. 10, inversion of the sequence order of two bases by accurate displacement vibration prediction can distinguish the blockade current and the vibration signal from the simultaneous measurement.

In this way, the sequence information obtained from the current waveform 403 is (CTGA GTGA CTGA) with respect to the base sequence order (CTGACTGA) of the original DNA 111, but this GTGA is read twice from the vibration prediction. It is possible to make a judgment, and by correcting this part, accurate sequence information can be obtained, so that the base sequence decoding accuracy can be improved.

As described above, according to the biological sample analysis method of the present embodiment, the device size can be reduced by vibration prediction without using an expensive device such as an active vibration isolator. Further, when vibration exceeding the control range of the active vibration isolation table occurs, the active vibration isolation table does not function, but according to the analysis method of this embodiment, the blocking current and the vibration signal are simultaneously measured to measure the vibration itself. When vibration is detected by the vibration detection unit and the reversal of the DNA sequence order is detected instead of suppressing the vibration, the ion current detected by the detection unit can be corrected after the measurement.

Further, it is generally said that the physical properties of DNA111 are not rigid but springy. Therefore, there is a possibility that the vibration and the behavior of the DNA between the thin film 101 in which the nanopores that are the detectors are opened and the driving component 109 that controls the passing speed of the DNA are different.

Therefore, the vibration between the thin film 101 in which the nanopores that are detectors are opened and the driving component 109 that controls the passage speed of DNA is predicted, and the springiness of DNA 111 is predicted by simulation to predict the behavior of DNA 111. , It is also possible to make corrections using this information.

FIG. 11 is a diagram for explaining a biological sample analyzer and a method according to Example 6. In general, it is considered that the measurement efficiency per thin film can be improved by using a so-called multi-nanopore in which a plurality of nanopores are provided on one nanopore thin film 1101. In this embodiment, the vibration detection unit detects vibration using an ion current for DNA of a known sequence, and the PC including the alarm unit analyzes the sequence information of DAN of an unknown sequence.

In the multi-pore system according to the biological sample analysis method 1100 of the present embodiment shown in FIG. 11, the accelerometer 501 and other speedometers and displacements are used for the vibration detection unit for detecting the vibration of the device on which the biological sample analyzer is mounted. You can think of a method that is not a measure. A plurality of DNA strands 1102 and 1103 are fixed to the DNA fixing substrate 108, and a plurality of nanopores of the multi-nanopore substrate 1101 are blocked. Of these, the DNA strand 1103 has a known base sequence.

Since the DNA sequencer is a device that analyzes unknown base sequence information, the base sequence order is reversed due to environmental vibration, and the base sequence decoding accuracy deteriorates. However, by simultaneously measuring the DNA 1103 whose base sequence is known and the DNA 1102 strand whose base sequence is unknown by the biological sample analysis method 1100, it is possible to detect the vibration by the known DNA strand 1103 when an unexpected vibration occurs. it can. Therefore, by correcting the unknown DNA strand 1102 by the detected vibration, it is possible to improve the base sequence decoding accuracy of the unknown DNA strand. Here, it is important that the known DNA strand 1103 is known, and other proteins, identification compounds, and the like may be used.

The present invention is not limited to the above-mentioned examples, and includes various modifications. For example, the above-mentioned examples have been described in detail for a better understanding of the present invention, and are not necessarily limited to those having all the configurations of the description. Further, it is possible to replace a part of the configuration of one embodiment with the configuration of another embodiment, and it is possible to add the configuration of another embodiment to the configuration of one embodiment. Further, it is possible to add / delete / replace a part of the configuration of each embodiment with another configuration.

Further, for each configuration, function, processing, etc. of the alarm unit and the like described above, an example of creating a program that realizes a part or all of them has been described, but a part or all of them is designed by, for example, an integrated circuit. Needless to say, it may be realized by hardware. That is, all or part of the functions of the processing unit may be realized by, for example, an integrated circuit such as an ASIC (Application Specific Integrated Circuit) or an FPGA (Field Programmable Gate Array) instead of the program.

100, 300, 500, 700, 800, 900 Biological sample analyzer 100A Partial enlarged view of biological sample analyzer 101: Partition 102A, 102B in which nanopore device is formed Tank 103 Electrolyte solution 104 Electrode 105A Power supply 105B Current meter 106 Flow cell 107 Probe 108 DNA fixing substrate 109 Drive mechanism 110 Flow cell opening 111 DNA
112 Nanopore 113 Nanopore thin film 200 Sequence reading example 301 Enlarged view of DNA111 302 Current waveform (no vibration)
401 Floor vibration 402 Acoustic vibration 403, 1000 Current waveform (with vibration)
501 Accelerometer 502 A / C Converter 503 PC
504 Alarm unit 600 Sequence reading example 801 Active vibration isolation table 802 Plate 803 Spring 804 Actuator 805, 901 Sensor 902 Displacement meter 903 Transfer function 1001 Enlarged view of nanopore device 1100 Biological sample analysis method 1101 Multi-nanopore substrate 1102 Unknown DNA
1103 Known DNA

Claims (11)

A bioanalyzer that analyzes the base sequence of DNA.
A nanopore substrate with nanometer-sized holes through which the object to be measured passes,
A tank whose inside is separated by the nanopore substrate and filled with an electrolyte solution,
A DNA fixing substrate that is located opposite to the nanopore substrate and on which the DNA is fixed,
The drive mechanism that drives the DNA fixing substrate and
A voltage application unit that applies voltage to both sides of the nanopore substrate,
A detector that detects the ion current when the DNA passes through the nanopore substrate, and
A vibration detection unit that detects the vibration of the bioanalyzer, and
A recording unit that records vibration information detected by the vibration detection unit,
It is provided with an alarm unit that issues an alarm when vibration is detected by the vibration detection unit.
The alarm unit controls so that the ion current detected while issuing an alarm is not used for nucleotide sequence decoding.
The alarm unit controls to stop the driving of the DNA fixing substrate by the driving mechanism while issuing an alarm.
A biological sample analyzer characterized by this. The biological sample analyzer according to claim 1 .
When the vibration detection unit detects that the vibration has subsided, the alarm unit restarts the driving of the DNA fixing substrate by the drive mechanism, and controls the detection unit to detect the ion current.
A biological sample analyzer characterized by this. The biological sample analyzer according to claim 1 .
When the vibration is detected by the vibration detection unit, the alarm unit feeds back the vibration information to the drive mechanism by a transfer function and drives the DNA fixing substrate so as to suppress the vibration of the DNA.
A biological sample analyzer characterized by this. The biological sample analyzer according to claim 1 .
When the vibration detection unit detects vibration and the DNA sequence reversal is detected, the alarm unit corrects the ion current detected by the detection unit.
A biological sample analyzer characterized by this. The biological sample analyzer according to claim 1 .
The nanopore substrate is provided with a plurality of nanometer-sized holes.
The DNA fixed substrate, and the DNA of the DNA and the known sequence of the unknown sequence Ru fixed,
A biological sample analyzer characterized by this. The biological sample analyzer according to claim 5 .
The vibration detection unit detects vibration by using the ion current with respect to the DNA of the known sequence.
A biological sample analyzer characterized by this. It is a bioanalytical method of a bioanalyzer that analyzes the base sequence of DNA.
The bioanalyzer has a nanometer-sized hole through which an object to be measured passes, and the inside is separated by a nanopore substrate to which a voltage is applied on both sides to face a tank filled with an electrolyte solution and the nanopore substrate. A DNA fixing substrate on which the DNA is fixed, a driving mechanism for driving the DNA fixing substrate, a detection unit for detecting an ion current when the DNA passes through the nanopore substrate, and the bioanalyzer. It is equipped with a vibration detection unit that detects the vibration information of
The base sequence is analyzed using the detected ion current and the detected vibration information.
An alarm is issued when the vibration information is detected, and the ion current detected when the alarm is issued is not used for base sequence decoding .
While the alarm is issued, the driving of the DNA fixing substrate by the driving mechanism is stopped.
A biological sample analysis method characterized by the above. The biological sample analysis method according to claim 7 .
When the vibration is detected based on the vibration information, the driving of the DNA fixing substrate by the driving mechanism is restarted, and the ion current is detected.
A biological sample analysis method characterized by the above. The biological sample analysis method according to claim 8 .
When the vibration information is detected and the sequence order of the DNA is reversed, the ion current detected by the detection unit is corrected.
A biological sample analysis method characterized by the above. It is a bioanalytical method of a bioanalyzer that analyzes the base sequence of DNA.
The bioanalyzer has a nanometer-sized hole through which an object to be measured passes, and the inside is separated by a nanopore substrate to which a voltage is applied on both sides to face a tank filled with an electrolyte solution and the nanopore substrate. A DNA fixing substrate on which the DNA is fixed, a driving mechanism for driving the DNA fixing substrate, a detection unit for detecting an ion current when the DNA passes through the nanopore substrate, and the bioanalyzer. It is equipped with a vibration detection unit that detects the vibration information of
The base sequence is analyzed using the detected ion current and the detected vibration information.
The nanopore substrate is provided with a plurality of nanometer-sized holes, and the DNA fixing substrate is used to fix an unknown sequence and a DNA having a known sequence.
A biological sample analysis method characterized by the above. The biological sample analysis method according to claim 10 .
The vibration detection unit detects vibration by using the ion current with respect to the DNA of the known sequence.
A biological sample analysis method characterized by the above.

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