JP2014020779A - Nucleic acid extraction device - Google Patents

Abstract

PROBLEM TO BE SOLVED: To achieve full automation including previous and subsequent processing operations such as carrying-in/out operations of a column or a container or the like to be used in a pressurization column type nucleic acid extraction device.SOLUTION: A nucleic acid extraction device includes: a dispensing mechanism 9 for dispensing a solution for nucleic acid extraction or the like; a gripper mechanism 8 for conveying and holding a column 4 and a container 6 to be used for nucleic acid extraction; and an XYZ arm mechanism 7 capable of moving those mechanisms 8 and 9 in an XYZ axial directions. Furthermore, the nucleic acid extraction device includes a column holder moving mechanism 30 for moving the column 4 between a column delivery/liquid injection position 22 and a column pressurizing position 23 while holding the column 4 by a column holder 3. The column moving mechanism 30 is constituted of a simultaneous two-axial operation mechanism, for example, a parallel link mechanism 31, for allowing the column holder 3 to simultaneously perform a biaxial action between the column pressurizing position 23 and the column delivery/liquid injection position 22 by one drive shaft.

Description

  The present invention relates to a nucleic acid extraction apparatus for extracting nucleic acid from a liquid sample containing nucleic acid using a column having a filter (nucleic acid binding solid phase).

  Conventional nucleic acid extraction methods include a magnetic bead method, a centrifugal method, and a method using a filter (for example, Patent Document 1 and Patent Document 2). In the filter-type nucleic acid extraction method, a pressure difference is generated between the inlet and the outlet of a column in which a filter composed of a nucleic acid-binding solid phase is mounted, and a liquid sample containing nucleic acid is prepared using the pressure difference. Through the filter, the nucleic acid is adsorbed to the filter. Thereafter, after a washing step in the column with a washing liquid, a liquid (so-called eluent) for removing the nucleic acid from the filter is passed through the filter, and the nucleic acid is separated from the filter and collected. As shown in Patent Documents 1 and 2, as a typical filter, a porous film of an organic material having a hydroxyl group on its surface, for example, a surface saponified product of acetylcellulose, is disclosed. As a porous film, for example, a glass filter containing a silica compound, which is an inorganic material having a hydrophilic group, is disclosed (Patent Document 2).

  One of nucleic acid extraction methods using a filter is a pressurized column method (Patent Document 2). In the pressurized column method, a liquid sample containing nucleic acid is injected into the column via a dispensing mechanism, and then a pressurized fluid (eg, pressurized air) is supplied to the column, and the liquid sample is passed through a filter. The washing liquid and the eluent are similarly passed through the filter using pressurized air.

  In the pressurized column type nucleic acid extraction apparatus in Patent Document 2, a pressurized air supply mechanism that is movable in the vertical direction is disposed above a column holder that holds a column (nucleic acid extraction cartridge). Below the column holder is disposed a container holder that can move in a direction (horizontal direction) just below the column holder, and a waste liquid container and a collection container are set in the container holder. In this nucleic acid extraction apparatus, when a liquid sample or washing liquid is passed through a column, a waste liquid container is located directly under the column. After the liquid sample or the cleaning liquid is supplied to the column via the dispensing mechanism, the pressurized air supply mechanism is lowered and the nozzle of the pressurized air supply mechanism is inserted into the upper opening of the column. In this state, by supplying pressurized air to the column, the liquid flows out of the column through the filter, and the effluent liquid (waste liquid) is received by the waste liquid container. When the eluent for removing nucleic acid is passed through the column, the movement of the container holder is controlled, and the collection container is positioned directly under the column. In the same manner as in the liquid sample supply and washing liquid supply steps described above, eluent injection and pressurized air are performed, and the liquid (eluent) containing nucleic acid flowing out from the column is recovered via the recovery container. Is done.

  In the nucleic acid extraction apparatus of Patent Document 2, a dispensing mechanism and a guide rail for moving the dispensing mechanism are arranged between the column holder and the pressurized air supply mechanism. When the column is mounted on the column holder, it is performed in a relatively narrow space between the column holder and the pressurized air supply mechanism, so it is performed through a hand, and the hand interferes with the dispensing mechanism. In order to avoid this, the dispensing mechanism is retracted from between the column holder and the pressurized air supply mechanism.

JP 2003-128691 A JP 2005-95113 A

  As described above, in the conventional pressure column type nucleic acid extraction apparatus, when the column is set in the column holder and when it is taken out after use, the processing by the user remains. Therefore, in the nucleic acid extraction process, if all the processes including pre- and post-processing operations other than pressurization are to be automated, it is difficult to realize with the conventional apparatus configuration.

  In particular, when trying to fully automate, the problem is that, in order to simplify the dispensing mechanism, if the sample is dispensed into the column using the dispensing mechanism, the column pressurizing mechanism and the dispensing mechanism are separated. It was difficult to realize due to interference with the injection mechanism.

  The present invention provides an apparatus that can realize full automation of the nucleic acid extraction process. In particular, a column pressurization mechanism and a dispensing mechanism can be automatically inserted and removed (positioning) at a column pressurization position, which conventionally requires manual processing, without requiring a hand. It is an object of the present invention to provide an apparatus that can eliminate the interference and fully automate the nucleic acid extraction process including before and after the pressurization process.

  The nucleic acid extraction apparatus of the present invention is basically configured as follows.

That is, a column having a filter for nucleic acid extraction;
A column holder for holding the column;
A pressurizing mechanism for applying pressure into the column;
A moving mechanism for a pressurizing mechanism that moves the pressurizing mechanism in the vertical direction;
A column holder moving mechanism that supports the column holder and moves the column holder between a column delivery / liquid injection position for performing column delivery and liquid injection into the column and a column pressurization position for applying pressure to the column; ,
A receiving container for receiving a liquid containing an extracted nucleic acid that leaves the column;
A receiving container transport mechanism for transporting the receiving container directly below the column pressure position;
A dispensing mechanism for inhaling and discharging various liquids used for nucleic acid extraction;
A gripper mechanism for gripping the column and receiving container;
A nucleic acid extraction apparatus comprising: a dispensing mechanism / a gripper mechanism moving mechanism capable of moving the dispensing mechanism and the gripper mechanism in at least three directions of an X axis, a Y axis, and a Z axis,
The column pressurization position is set immediately below the pressurization mechanism, and the column delivery / liquid injection position where the column is delivered is a position deviating from just below the pressurization mechanism, the X axis, the Y axis, and the Z axis Together with the column pressure position on the biaxial surface of any of
The column holder moving mechanism is configured by a simultaneous biaxial operation mechanism that moves the column holder between the column delivery / liquid injection position and the column pressurization position by a simultaneous biaxial operation with a single drive shaft. It is characterized by being.

  With the above configuration, the nucleic acid extraction apparatus of the present invention can fully automate the extraction process of the pressurized column system while reducing contamination and contamination risk.

  As a result, it is possible to improve the inspection efficiency by releasing from manual work and improving the result defect due to work mistakes.

1 is a plan view showing an overall outline of a nucleic acid extraction apparatus according to Embodiment 1 of the present invention. The top view which shows the periphery of the pressurization mechanism used in order to perform the nucleic acid extraction in the said nucleic acid extraction apparatus. The front view which shows the periphery of the pressurization mechanism used in order to perform the nucleic acid extraction in the said nucleic acid extraction apparatus. The side view which shows the periphery of the pressurization mechanism used in order to perform the nucleic acid extraction in the said nucleic acid extraction apparatus. Explanatory drawing which shows an example of the column holder moving mechanism for hold | maintaining and moving the column holder in the said nucleic acid extraction apparatus. Explanatory drawing which shows the other example of the column holder moving mechanism for hold | maintaining and moving the column holder in the said nucleic acid extraction apparatus. Explanatory drawing which shows the other example of the column holder moving mechanism for hold | maintaining and moving the column holder in the said nucleic acid extraction apparatus. Explanatory drawing which shows the other example of the column holder moving mechanism for hold | maintaining and moving the column holder in the said nucleic acid extraction apparatus. The top view which shows the whole outline | summary of the nucleic acid extraction apparatus which concerns on Example 2 of this invention.

  Hereinafter, embodiments of the present invention will be described with reference to examples and the drawings.

  FIG. 1 is a top view showing the overall configuration of a nucleic acid extraction apparatus according to Embodiment 1 of the present invention, and FIGS. 2A, 2B and 2C show the periphery of a pressurizing mechanism used for nucleic acid extraction in the nucleic acid extraction apparatus. FIG. 3 is an explanatory view showing an example of a column holder moving mechanism for holding and moving the column holder used in the first embodiment. FIG. 3 is a front view of the column holder mechanism.

  In FIG. 1, a nucleic acid extraction apparatus 1 according to the present embodiment includes a pressurizing mechanism 2 for supplying a pressurized fluid such as pressurized air to a column 4 for nucleic acid extraction, and a column holder 3 for holding the column 4. A column holder moving mechanism 30 (illustrated in FIG. 3) that moves while supporting the column holder 3 and a receiving container (extracted nucleic acid receiving container) 6 for receiving a liquid containing the extracted nucleic acid that is detached from the column 4 are conveyed. And a receiving container transport mechanism 5. Details of these mechanisms will be described later.

  The nucleic acid extraction apparatus 1 holds a dispensing mechanism 9 and a gripper mechanism 8, and moves these mechanisms in the X-axis, Y-axis, and Z-axis directions (dispensing mechanism / moving mechanism for gripper mechanism). 7 is provided. The XYZ arm mechanism 7 includes a Y-axis arm 7a that operates in the Y-axis direction, X-axis arms 7b-1 and 7b-2 that are attached to the Y-axis arm 7a so as to be movable in the X-axis direction, and a movement in the Z-axis direction. A gripper mechanism 8 attached to the X-axis arm 7b-1 so as to be able to be provided, and a dispensing mechanism 9 attached to the X-axis arm 7b-2 so as to be movable in the Z-axis direction are provided. Note that the XYZ arm mechanism 7 itself is a well-known mechanism, and is schematically illustrated here. In FIG. 1, for simplification of the drawing, the X-axis arm 7 b-1 and the gripper mechanism 8, and the X-axis arm 7 b-2 and the dispensing mechanism 9 are shown as common blocks. , Each is composed of independent elements. The XYZ arm mechanism 7 includes an actuator (not shown) for movement in each axial direction.

  The nucleic acid extraction apparatus 1 includes a receiving container rack 16 for storing 6 groups of unused receiving containers (extracted nucleic acid receiving containers) within a movable range of the XYZ arm mechanism 7 and a column magazine 14 for storing 4 groups of unused columns. A column magazine setting section (column group setting section) 14 'for setting a sample container and a specimen container rack for setting a specimen container (pretreatment container for nucleic acid extraction) 13 containing a specimen (sample) for nucleic acid extraction 12 (a pretreatment container installation part), a dispensing chip magazine set part (dispensing chip group installation part) 10 ′ for setting a dispensing chip magazine 10 for storing unused dispensing chips 11 groups, Reagent container setting part (reagent container installation part) 17 'for setting a reagent container 17 containing various reagents necessary for nucleic acid extraction, used column, used dispensing tip, used receiving A reject bin 24 for discarding vessels, collecting container setting portion comprises a collection container 25 group for recovering nucleic acids already extracted liquid (recovery container installation section) 25 'are disposed. The column 4 is equipped with a filter made of a nucleic acid-binding solid phase. Since the material of the filter is well known in Patent Documents 1 and 2 as described above, the description thereof is omitted.

  Here, an example of the column holder moving mechanism 30 will be described with reference to FIG. The column holder moving mechanism 30 is constituted by, for example, a parallel link mechanism 31 and includes a pair of parallel link elements 31a and 31b. One end (lower end) of each of the parallel link elements 31a and 31b is supported by the fixed base 34 via a drive shaft 32a and a driven shaft 32b, respectively, and is rotatably attached. The drive shaft 32a is rotated by an actuator (motor) 33, and the driven shaft 32b is rotated through a link by the rotation. The other ends (upper ends) of the parallel link elements 31 a and 31 b are connected to each other via the column holder 3. The parallel link elements 31a and 31b are rotated within a predetermined range by the actuator 33, whereby the column holder 3 performs an arc motion between two points.

  Of the two arc movements of the column holder 3, one point is located near the pressurizing mechanism 2 and in front of the pressurizing mechanism 2 (position 22 in FIGS. 1 and 2C). When the gripper mechanism 8 that grips the unused column 4 immediately above the position 22 is positioned by the XYZ arm mechanism 7, the gripper mechanism 8 is opened and the column 4 is transferred from the gripper mechanism 8 to the column holder 3. When the dispensing mechanism 9 is positioned by the XYZ arm mechanism 7 immediately above the column 4 at the column delivery / liquid injection position 22, the liquid from the dispensing mechanism 9 is set to be supplied to the column 4. The other point is located near the pressurizing mechanism 2 and immediately below the pressurization mechanism 2 (column pressurization position 23). It is comprised so that it may be pressurized with pressurized air.

  As shown in FIGS. 2B and 2C, the receiving container transport mechanism 5 includes a holder 60 for holding the receiving container 6, a holder standby position 21 for delivering the receiving container (extracted nucleic acid receiving container) 6, and the pressurizing mechanism 2. Are moved between a position 20 immediately below the column pressurization position 23 (position just below the pressurization) and a liquid intake position 19 for sucking a liquid (a liquid containing an extracted nucleic acid as will be described later) in the receiving container 6. is there. The receiving container transport mechanism 5 is configured by an appropriate linear motion mechanism such as a belt conveyor mechanism or a pinion rack mechanism capable of reciprocating. In addition, the receiving container transport mechanism 5 is not limited to a linear motion mechanism, and can be configured by a rotational motion mechanism such as a rotary table.

  When performing the nucleic acid extraction operation, the user sets a sample container (nucleic acid extraction pretreatment container) 13 containing a sample solution (sample) containing nucleic acid in advance in the sample container rack 12, and the dispensing tip magazine 10 and The column magazine 14 and the reagent container 17 are set at predetermined positions, and the receiving container 6 is set on the receiving container rack 16.

  When the start button is pressed, the following series of operations are performed by process control of a controller (not shown). The XYZ arm mechanism 7 positions the dispensing mechanism 9 directly above the dispensing tip magazine 10, the dispensing mechanism 9 descends at this position, and the dispensing tip 11 is attached to the dispensing mechanism 9.

  After the dispensing tip 11 is mounted, the dispensing mechanism 9 moves to the predetermined reagent container 17 via the XYZ arm mechanism 7, and the dispensing tip 11 aspirates the reagent. Thereafter, the dispensing mechanism 9 moves to the position of the sample container rack 12 via the XYZ arm mechanism 7 and discharges (dispenses) the reagent to the sample container 13 there. After the dispensing, the dispensing tip 11 moves up and down together with the dispensing mechanism 9 to stir the solution (sample liquid and reagent) in the sample container 13. Instead of directly discharging the reagent into the specimen container 13, another container (not shown) may be prepared, and the reagent and specimen may be dispensed and mixed there.

  While the dispensing mechanism 9 is performing a series of steps (so-called nucleic acid extraction pretreatment step) from mounting the dispensing tip to stirring the solution in the specimen container 13 as described above, the column holder 3 is at the column delivery / liquid injection position 22. Waiting.

  After the solution is stirred by the dispensing mechanism 9, the gripper mechanism 8 moves to the position of the column magazine 14 via the XYZ arm mechanism 7, and the gripper mechanism 8 grips the column 4 from the column magazine 14. Thereafter, the column 4 held by the gripper mechanism 8 is transported to the column holder 3 at the column delivery / liquid injection position 22 via the XYZ arm mechanism 7 and delivered to the column holder 3.

  After delivery of the column 4, the dispensing mechanism 9 moves to a predetermined sample container 13 position via the XYZ arm mechanism 7. At the position of the specimen container 13, the dispensing mechanism 9 sucks the solution (that is, a mixed solution of a sample containing nucleic acid and a reagent). After sucking the solution, the dispensing mechanism 9 moves to the position immediately above the column holder 3 waiting at the column delivery / liquid injection position 22 via the XYZ arm mechanism 7, and the solution is discharged from the dispensing mechanism 9 to the column 4. To do.

  After discharging the solution, the actuator 33 of the column holder moving mechanism 30 is driven (for example, counterclockwise in FIG. 3), and the parallel link elements 31a and 31b are rotated counterclockwise with the drive shaft 32a and the driven shaft 32b as fulcrums. Rotate the set angle. The set rotation angle of the parallel link element is an angle between the column delivery / liquid injection position 22 and the column pressurization position 23 with reference to the fulcrum of the parallel link. Due to the rotation of the parallel link element, the column holder 3 moves to a column pressurization position 23 that is a position immediately below the pressurization mechanism 2 by an arc motion via the parallel link mechanism. This movement requires two-axis movement in the horizontal direction (Y-axis direction) and the vertical direction (Z-axis direction). However, the movement between the two points via the parallel link mechanism (rotation operation within a predetermined range) By doing so, it is possible to simultaneously perform the movement in the two axial directions with one actuator.

  When the column holder 3 moves to the column pressurizing position 23, the holder 60 of the receiving container transport mechanism 5 in which the receiving container 6 is set is in the standby position 21.

  After the column holder 3 has moved to the column pressurization position 23, the pressurization mechanism 2 is lowered, and a part of the nozzle portion of the pressurization mechanism 2 comes into close contact with the upper end portion of the column 4 in the column holder 3. The method of close-contact is not limited as long as close contact such as compression by O-ring (not shown) or insertion is achieved.

  After the close contact, the pressurizing mechanism 2 sends a pressurized fluid (for example, pressurized air) to the column 4 to perform a pressurizing operation. By this pressurizing operation, the solution contained in the column 4 is pushed by pressure, the filter of the column 4 is passed through, and the waste liquid is discharged from the lower part of the column. The discharged waste liquid flows into the waste liquid receiver 18 and is stored. The stored waste liquid is collected in the waste liquid receiver 18 and stored in a bottle or the like. When the pressurization is finished, the pressurization mechanism 2 moves upward and moves away from the upper end of the column 4. The waste liquid receiver 18 is provided so as to cover the entire area immediately below the range in which the column holder 4 performs an arc motion. Accordingly, even if the solution in the column 4 is spilled while the column 4 is moving between the column delivery / liquid injection position 22 and the column pressurization position 23 via the column holder 3. Since it is collected in the waste liquid receiver 18, contamination can be prevented.

  When the first pressurization is finished, the column holder 3 then returns from the column pressurization position 23 to the column delivery / liquid injection position 22 by the simultaneous biaxial operation via the column holder moving mechanism 30. Thereafter, the process proceeds to the step of pressurizing and passing a reagent such as a washing solution through the column 4, but from the dispensing tip mounting to the pressurization to the column 4 is performed by the same method as the above solution.

  After the series of steps, an eluent is supplied to the column 4 as a reagent in order to release the nucleic acid adsorbed on the filter of the column 4. From the dispensing tip mounting to the injection of the eluent into the column 4 is performed in the same manner as the process from the dispensing tip mounting to the liquid delivery at the column delivery / liquid injection position 22 such as the above-described solution and washing solution. .

  Thereafter, the gripper mechanism 8 moves to the receiving container rack 16 via the XYZ arm mechanism 7, and the gripper mechanism 8 takes out the receiving container 6 from the rack 16. Thereafter, the gripper mechanism 8 moves to the holder standby position 21 on the receiving container transport mechanism 5, and sets the receiving container 6 in the holder 60 at the standby position 21. When the receiving container 6 is set in the holder 60, the receiving container 6 moves together with the holder 60 to a position 20 immediately below the column pressurizing position 23 via the receiving container transport mechanism 5.

  Under this state, the column holder 3 moves in a circular arc (simultaneous biaxial operation) via the column holder moving mechanism 30 and moves to the column pressurizing position 23. By this movement, the lower end portion of the column 4 held by the column holder 3 is inserted into the receiving container 6 of the receiving container transport mechanism 5. In this state, the column 4 and the receiving container 6 are in a positional relationship where they do not come into contact. This is to prevent contamination.

  Thereafter, the pressurizing operation is started by the pressurizing mechanism 2 as before. The passed liquid (eluent) is a liquid containing nucleic acid (extracted nucleic acid) released from the filter, and is all collected in the receiving container 6.

  Thereafter, the column holder 3 returns to the column delivery / liquid injection position 22 via the column holder moving mechanism 30, and the used column 4 is gripped by the gripper mechanism 8 and moved to the waste box 24 via the XYZ arm mechanism 7. Then, the column 4 is discarded to the disposal box 24. On the other hand, the holder 60 of the receiving container transport mechanism 5 moves to the liquid intake position 19 with the receiving container 6. In this state, the dispensing mechanism 9 moves to the liquid intake position 19 via the XYZ arm mechanism 7, and the dispensing mechanism 9 sucks the liquid containing the extracted nucleic acid from the receiving container 6. Thereafter, the dispensing mechanism 9 moves to the position of the collection container 25 via the XYZ arm mechanism 7, and the liquid containing the extracted nucleic acid is moved to the collection container 25 via the dispensing mechanism 9. Thereafter, the used receiving container 6 is moved from the liquid intake position 19 to the disposal box 24 via the XYZ arm mechanism 7 and the gripper mechanism 8 and discarded.

  Through the above-described series of operations, the fully automated process of the extraction process is completed.

  During operation, liquid dispensing / dispensing (dispensing operation) by the dispensing mechanism 9 is performed when a different type of sample, reagent, etc. is handled, and a new dispensing tip is reattached each time. The chip is discarded in the disposal box 24.

  In the said Example, you may provide the XYZ arm mechanism 7, the gripper mechanism 8, the dispensing mechanism 9, and the pressurization mechanism 2 with two or more. Moreover, the structure provided with two or more containers supported by the column holder 3 or the receiving container conveyance mechanism 5 may be sufficient. In particular, when the column holder 3 and the receiving container transport mechanism 5 are provided with a plurality of containers, the risk of contamination between adjacent containers can be reduced by using the mechanism configuration and operation method of this embodiment.

  According to the present embodiment, it is possible to enable full automation including front and rear processing operations such as loading and unloading operations of columns and containers used in the nucleic acid extraction apparatus. In addition, even when the waste liquid drips from the column, the apparatus can be prevented from being contaminated. In addition, since the column holder is operated by a single actuator, smooth operation can be realized, and after dispensing the solution to the column, the risk of spilling the solution while transporting the column can be reduced, and contamination risk can be reduced. is there.

For these reasons, it is possible to provide a nucleic acid extraction apparatus that fully automates the pre- and post-treatment while reducing contamination and contamination risk.

  The column holder moving mechanism 30 may be a simultaneous biaxial operation mechanism that simultaneously performs biaxial operation between the column delivery / liquid injection position 22 and the column pressurization position 23 with a single drive shaft, and various forms are conceivable.

  FIG. 4 shows another example of the column holder moving mechanism 30. FIG. 4 is a front view of the column holder mechanism 30. In this example, the simultaneous biaxial operation mechanism 31 constituting the column holder moving mechanism 30 includes a drive shaft 43 that performs linear motion, and parallel links 31a and 31b that convert this linear motion into a predetermined range of rotational motion. The free ends of the parallel links 31 a and 31 b are connected via the column holder 3. More specifically, each of the parallel links 31 a and 31 b is supported by a cam shaft 45 whose one end converts a linear motion into a rotational motion, and each cam shaft 46 is engaged with a cam groove 46. Each cam shaft 46 is coupled to an output shaft of an actuator 43 that performs a linear operation such as a solenoid. The middle of the parallel links 31a and 31b is supported by the fixed base 44 via a shaft 42 which is a fulcrum of the parallel link. In the column holder moving mechanism 30 of this example, when the output shaft of the actuator 43 performs a reciprocating motion linearly, the parallel links 31 a and 31 b rotate within the cam groove 46. This rotation operation is performed within the range between the column pressurizing position 23 and the column delivery / liquid injection position 22 as in the column holder moving mechanism of FIG. As a result, the column holder 3 performs an arc motion (simultaneous biaxial motion of the Y axis and Z axis) within the range between the column pressurization position 23 and the column delivery / liquid injection position 22.

  FIG. 5 shows another example of the column holder moving mechanism 30. FIG. 5 is a top view of the column holder moving mechanism 30 of this example.

  The column holder moving mechanism 30 of this example has the same components as those in FIG. 3, but the operation direction of the parallel links is different. That is, the column holder moving mechanism 30 in FIG. 3 rotates on the surfaces in the vertical direction (Z-axis direction) and the horizontal direction (Y-axis direction), whereas in this example, two horizontal axes (X (Axis, Y-axis). The column pressurization position 23 and the column delivery / liquid injection position 22 which are the operating points of the parallel link mechanism 31 are both on the X-axis and Y-axis planes.

  Also in this example, one end of each of the parallel link elements 31a and 31b is supported by the fixed base 34 via the drive shaft 32a and the driven shaft 32b, respectively, and is rotatably attached. The drive shaft 32a is rotated by an actuator (motor) 33, and the driven shaft 32b is rotated through a link by the rotation. The other ends (upper ends) of the parallel link elements 31 a and 31 b are connected to each other via the column holder 3. The parallel link elements 31a and 31b are arranged at a height position between the receiving container transport mechanism 5 and the pressurizing mechanism 2, and are rotated by the actuator 33 on the horizontal plane (X-axis Y-axis plane) at this height position. A column pressurizing position 23 and a column delivery / liquid injection position 22 are arranged on the horizontal plane.

  In the example of FIG. 3, the column holder 3 performs an arc motion in the Y-axis and Z-axis directions. Therefore, as shown in FIG. 2C, when the column holder 3 comes to the column pressurization position 23, the column holder 3 naturally moves to the receiving container 6. Although the tip is inserted, in FIG. 5, the column holder 3 performs an arc motion in the X-axis and Y-axis directions. Therefore, such an operation cannot be expected, and the column holder 3 is moved to the column pressurization position 23. When it comes, a part of the column 4 is directly above the receiving container 6 (not inserted). However, since the parallel link elements 31a and 31b are cantilevered by the shafts 32a and 32b, the nozzle of the pressurizing mechanism 2 is attached to the column 4 by using a member having elasticity (flexibility). If the nozzle is pushed downward by this nozzle when inserted, the tip of the column 4 can be inserted into the receiving container 6.

  FIG. 6 shows another example of the column holder moving mechanism 30. FIG. 6 is a top view of the column holder moving mechanism 30 of this example.

  The column holder moving mechanism 30 of this example has the same components as those in FIG. 4, but the operation direction of the parallel links is different. That is, the column holder moving mechanism 30 in FIG. 4 rotates on the surfaces in the vertical direction (Z-axis direction) and the horizontal direction (Y-axis direction), whereas in this example, as in the example in FIG. It is configured to rotate on the surface of two horizontal axes (X-axis and Y-axis).

  In the column holder moving mechanism 30 of this example, the parallel links 31 a and 31 b rotate within the cam groove 46 when the output shaft of the actuator 43 reciprocates linearly, as in FIG. Also in this example, the parallel link elements 31a and 31b are disposed at a height position between the receiving container transport mechanism 5 and the pressurization mechanism 2, and the actuator 43 is set to a horizontal plane (X-axis Y-axis plane) at this height position. The column pressurization position 23 and the column delivery / liquid injection position 22 are arranged on the horizontal plane. The mechanism described in the example of FIG. 5 in which a part of the column 4 at the column pressurizing position 23 can be inserted into the receiving container 6 can be applied to the column holder mechanism 30 of FIG.

  FIG. 7 is a plan view showing an overall outline of the nucleic acid extraction apparatus according to Embodiment 2 of the present invention. In this example, an apparatus configuration in which the nucleic acid extraction apparatus of Example 1 and an amplification detection apparatus are combined is shown.

  In Example 1, the extracted nucleic acid sample was collected in the collection container 25, but in this example, it is dispensed in the reaction container 50 for nucleic acid amplification instead of the collection container 25. Thereafter, the dispensing mechanism 9 dispenses the reagent used for amplification from the pretreatment reagent for amplification 26 to the reaction container 50 with a predetermined movement operation by the XYZ arm mechanism 7. Thereafter, after performing a dispensing and stirring operation and adding a reagent as necessary, the reaction vessel 50 is transported and installed from the reaction vessel magazine 51 to the closing mechanism 27 by the gripper mechanism 8 of the XYZ arm mechanism 7.

  The reaction vessel 50 is closed by the closing mechanism 27 and sealed so that the reaction solution does not leak. Thereafter, the reaction vessel 50 is unloaded from the closing mechanism 27 by the gripper mechanism 8 of the XYZ arm mechanism 7 and is transported and set to the stirring mechanism 28.

  Thereafter, a stirring operation is performed by the stirring mechanism 28 to stir the reaction solution in the reaction vessel 50. After the stirring operation, the gripper mechanism 8 of the XYZ arm mechanism 7 conveys and sets the amplification detection mechanism 29.

  The amplification detection mechanism 29 performs amplification and fluorescence measurement by applying temperature, and outputs the result. Amplification prefectures and fluorescence measurement are well-known technical matters, and therefore their explanation is omitted. The configuration of the amplification detection mechanism 29 may be either a mechanism corresponding to the batch processing method or a mechanism corresponding to the random access method.

  With the above-described operation, the extraction process and the amplification detection process can be performed fully automatically.

DESCRIPTION OF SYMBOLS 1 ... Nucleic acid extraction apparatus, 2 ... Pressurization mechanism, 3 ... Column holder, 4 ... Column, 5 ... Reception container conveyance mechanism, 6 ... Reception container, 7 ... XYZ arm mechanism, 8 ... Gripper mechanism, 9 ... Dispensing mechanism, DESCRIPTION OF SYMBOLS 10 ... Dispensing tip magazine, 11 ... Dispensing tip, 12 ... Sample container rack, 13 ... Sample container, 14 ... Column magazine, 15 ... Column, 16 ... Receiving container rack, 17 ... Reagent container, 18 ... Waste liquid receptacle, 19 DESCRIPTION OF REFERENCE SIGNS ... Transport mechanism liquid take-in position, 20 ... Position just below column pressurization, 21 ... Transport mechanism holder standby position, 22 ... Column delivery / liquid injection position, 23 ... Column pressurization position, 24 ... Waste box, 25 ... Extracted nucleic acid collection container , 26 ... Pretreatment reagent for amplification, 27 ... Closing mechanism, 28 ... Stirring mechanism, 29 ... Amplification detection mechanism, 50 ... Reaction vessel, 51 ... Reaction vessel magazine.

Claims (10)

A column having a filter for nucleic acid extraction;
A column holder for holding the column;
A pressurizing mechanism for applying pressure into the column;
A moving mechanism for a pressurizing mechanism that moves the pressurizing mechanism in the vertical direction;
A column holder moving mechanism that supports the column holder and moves the column holder between a column delivery / liquid injection position for performing column delivery and liquid injection into the column and a column pressurization position for applying pressure to the column; ,
A receiving container for receiving a liquid containing an extracted nucleic acid that leaves the column;
A receiving container transport mechanism for transporting the receiving container directly below the column pressure position;
A dispensing mechanism for inhaling and discharging various liquids used for nucleic acid extraction;
A gripper mechanism for gripping the column and receiving container;
A nucleic acid extraction apparatus comprising: a dispensing mechanism / a gripper mechanism moving mechanism capable of moving the dispensing mechanism and the gripper mechanism in at least three directions of an X axis, a Y axis, and a Z axis,
The column pressurization position is set immediately below the pressurization mechanism, and the column delivery / liquid injection position where the column is delivered is a position deviating from just below the pressurization mechanism, the X axis, the Y axis, and the Z axis Together with the column pressure position on the biaxial surface of any of
The column holder moving mechanism is configured by a simultaneous biaxial operation mechanism that moves the column holder between the column delivery / liquid injection position and the column pressurization position by a simultaneous biaxial operation with a single drive shaft. A nucleic acid extraction apparatus characterized by comprising:   The nucleic acid extraction apparatus according to claim 1, wherein the column delivery / liquid injection position is set at a position in front of the pressurizing mechanism and obliquely above the column pressurizing position.   A pretreatment container installation portion in which a sample for nucleic acid extraction to be subjected to nucleic acid extraction pretreatment is placed in an area where the movement mechanism for the dispensing mechanism / gripper mechanism is movable; and a reagent container installation portion for containing a reagent; The installation section of the column group before use, the column delivery / liquid injection position, the installation section of the reception container group before use, and the standby position of the reception container holder of the reception container transport mechanism are set. Item 3. The nucleic acid extraction apparatus according to Item 1 or 2.   The simultaneous two-axis operation mechanism is a rotating shaft in which the driving shaft is rotated by a motor, and includes a parallel link that rotates in a predetermined range around the driving shaft and the driven shaft, and the free ends of the parallel links are The nucleic acid extraction apparatus according to any one of claims 1 to 3, wherein the nucleic acid extraction apparatus is connected via the column holder.   The simultaneous biaxial operation mechanism includes a drive shaft that performs a linear motion, and a parallel link that converts the linear motion into a rotational motion within a predetermined range, and free ends of the parallel links are connected to each other via the column holder. The nucleic acid extraction apparatus according to any one of claims 1 to 3, which is connected.   The moving range of the receiving container transport mechanism includes a position for delivering the receiving container, a position immediately below the column pressurizing position, and a dispensing position for sucking the liquid in the receiving container. The nucleic acid extraction apparatus according to any one of 1 to 5.   A waste liquid receiver is disposed below the column pressurization position, and when the receiving container is located at a position other than the column pressurization position, the waste liquid from the column holder flows down directly to the waste liquid receiver. Item 7. The nucleic acid extraction device according to any one of Items 1 to 6.   The nucleic acid extraction apparatus according to claim 7, wherein the waste liquid receiver is provided so as to cover the entire area immediately below the range in which the column holder moves.   When the column holder moving mechanism is simultaneously biaxially operated from the column delivery / liquid injection position to the column pressurization position while the column is held by the column holder, the tip of the column is inserted into the receiving container. The nucleic acid extraction apparatus according to any one of claims 1 to 8, wherein a movement range of the column holder movement mechanism is set.   The pressurizing mechanism is set so that the outer wall of the nozzle part of the pressurizing mechanism is brought into close contact with the upper inner periphery of the column in a state where the tip of the column is inserted into the receiving container. Item 10. The nucleic acid extraction device according to Item 9.

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