JP2009168734A - Liquid tester - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a liquid tester for preventing liquid flow from one container to another container when liquid levels of the containers are different and preventing invasion of foreign objects into liquid in the containers. <P>SOLUTION: The liquid tester includes a plurality of pipette chips 101, a plurality of ducts 103, a fan 106, a suction member 107, and a control portion 200. The duct 103 isolates one pipette chip 101 and the inside of a container corresponding to the pipette chip 101 from the other pipette chip 101 and the insides of the other containers. Each pipette chip 101 is so constructed to move relative to the duct 103. The fan 106 is communicated with the inside of the duct 103 to apply a pressure to the inside of the duct 103. When at least the pipette chip 101 starts to discharge, the control portion 200 moves the duct 103 until it touches the container and drives the fan 106. <P>COPYRIGHT: (C)2009,JPO&INPIT

Description

  The present invention relates to a liquid inspection apparatus that prevents inadvertent mixing of substances to be inspected used when performing a solution operation with a pipette.

  Conventionally, the DNA testing process is performed manually and requires a great amount of labor, which may cause an adjustment error, and requires skill. In order to solve these problems, Patent Document 1 discloses an automatic apparatus that consistently performs a process from extraction of a nucleic acid from a biological sample to detection. This automatic device consists of a dispenser, a warmer, a cold room, a detector, a transporter, a controller, etc., and the container moves the container between the dispenser, warmer, cooler, and detector by the transporter. It has become.

  In an automatic apparatus that handles nucleic acids as described in Patent Document 1, when handling a plurality of test objects at the same time, care must be taken not to inadvertently mix each other's nucleic acids.

  The dispenser that dispenses samples and reagents configured in this automatic apparatus has the following problems.

When the solution sucked into the pipette tip attached to the dispenser is discharged to another container, the solution may float in the atmosphere as follows.
・ Liquid with bubbles formed on the tip of the pipette tip or the liquid surface when it is discharged or a solution that is torn off from the main drop ・ Solution that is mist-like from the tip of the pipette tip when discharged ・ Solution discharged from the pipette tip The solution that bounces by hitting the liquid surface or the inner wall of the container already stored. When stirring a plurality of solutions in one container, the solution may float in the atmosphere as follows.
・ Mist-like solution from the liquid level due to stirring ・ Bubble formed on the liquid surface due to stirring and then splashed.Furthermore, when the pipette tip enters and exits the solution in the container, The generated droplet, mist, and the volume of air that has been pushed away when the pipette tip enters the container may move in the container opening direction, and the mist in the air may be scattered outside and float.

  There is a possibility that such floating substances may be mixed in another adjacent container or floating in the apparatus and mixed in a container used for the next analysis or inspection. When nucleic acids are contained in these floating substances, if floating substances from a container adjacent to the container to be amplified are mixed, the number of nucleic acids that should not exist is increased exponentially, and the accuracy of the test results is increased. Remarkably decreases the performance.

Patent Documents 2 and 3 disclose methods for solving these problems.
JP-A-7-107999 JP-A-2-31165 JP 2006-158335 A

  In Patent Document 2, it is described as a problem that a sample that has become mist at the time of discharge is scattered around and is secondarily infected to an operator. In order to prevent this, in Patent Document 2, gas suction means is provided in the vicinity of the dispensing means. Specifically, the duct as the suction means is disposed at a position slightly away from the liquid discharge port of the probe (pipette tip) and at a position where the atomized gas can be sucked from obliquely above in the discharge direction.

  The problem with this configuration is that the arrangement of the probe and duct is limited. When the container is thin and deep, it is necessary to penetrate the probe into the container in order to discharge the solution. In order to cope with this, it is necessary to arrange the duct so that it does not hit the container, and the distance between the tip of the probe and the duct is increased. There is a possibility that it cannot be recovered sufficiently.

  Therefore, it is necessary to take measures such as increasing the suction force, that is, increasing the pressure generating means such as a fan, or improving the driving force. However, there arises a problem that the operation sound of the apparatus increases.

  In addition, when the present inventor observed the periphery of the tip of the pipette tip at the time of discharge, a droplet of about 10 μm that seems to have been torn off from the discharged main droplet was in a direction substantially perpendicular to the discharge direction at an initial speed of about 100 mm / second, In other words, it was seen to fly almost horizontally.

  These droplets may fly in any direction of 360 °, and in order to collect them, ducts must be arranged around the entire pipette tip, the pitch between adjacent containers is increased, and the apparatus is large. It will become.

  In addition, the device disclosed in Patent Document 1 tries to recover the inside of the cover by sucking it from the duct while preventing the liquid droplets from being scattered outside, but depending on the shape and arrangement of the components in the device. The airflow may not be formed in the desired direction.

  In particular, in an apparatus that processes multiple specimens, that is, specimens that should not be mixed in an adjacent positional relationship, the mist-like specimen may adhere to the probe of another specimen before it reaches the duct when aspirated. There is. If the probe is re-entered into the container with the mist-like sample attached to the probe of another sample, there is a high possibility that the other sample will be mixed in the container and unnecessary reaction will start.

  Furthermore, in order to suck a wide area inside the cover, the suction force is increased and a large amount of air is sucked, so the amount of dust and foreign matter inside and outside the device contained in the suction air also increases. In addition, there is a possibility of adhering when passing around the container. If such dust and foreign matter are mixed in the solution, another problem of inhibiting the reaction occurs.

  Patent Document 3 describes a method of processing splashes when a liquid is discharged from a dispensing port, but has the following problems.

  Since the suction port and the jet port are formed integrally with the tip for manipulating the liquid, a general-purpose pipette tip cannot be used and a running cost is required. In addition, since the suction port and the jet port are formed integrally with the chip for manipulating the liquid, it is difficult to pipette in a thin and deep container such as a commercially available PCR plate. If the entire chip is thinned to cope with a thin container, the droplet processing performance is lowered, and the droplets that could not be processed are scattered outside the container.

  In addition, since the suction port, the spout and the dispensing port are close in the height direction, when the dispensing port is plunged into the solution and discharged or sucked, depending on the depth of penetration, the suction port and the spout May come into contact with the liquid surface. As a result, there is a possibility that a film is formed at the suction port, which is broken and becomes mist, or extra mist is generated near the jet port.

  The height relationship between the dispensing port, the suction port, and the ejection port may be determined so that this does not occur, but it cannot cope with the case where the relationship between the dispensing port and the liquid surface height varies relatively. For example, when a plurality of liquid volumes are set for one container, in order to operate the solution by inserting a chip into the liquid, the suction port and the jet are also applied to the highest liquid level. The positional relationship with the dispensing port must be determined so that the outlet does not touch. Then, when operating without inserting the tip into the liquid, the suction port and jet port are far from the liquid surface, and the liquid droplets flying in the horizontal direction from the vicinity of the dispensing port as described above are processed. Becomes very difficult.

  A common problem in the devices disclosed in Patent Document 2 and Patent Document 3 is that there is a limit to the droplet processing capacity when handling a solution by inserting a chip into a thin container or inserting a chip into a liquid. It can be said that there is a point. In other words, the devices disclosed in Patent Document 2 and Patent Document 3 are based on the premise that the positional relationship between the liquid level and the tip of the chip is limited to a certain range.

  Therefore, the present invention can prevent the liquid in one container from mixing into the liquid in another container even if the height of the liquid level in the container fluctuates, and the liquid in the container can have foreign matters. It is an object of the present invention to provide a liquid inspection apparatus that can prevent mixing.

In order to achieve the above object, the liquid inspection apparatus of the present invention includes a plurality of liquid discharge suction means for discharging the liquid into the container and sucking the liquid held in the container,
A plurality of shielding means for shielding the inside of the container corresponding to the liquid discharge suction means and the liquid discharge suction means from the other liquid discharge suction means and the inside of the other container;
A pressure generating means communicating with the shielding means and applying pressure to the inside of the shielding means;
An absorbent member that is disposed inside the shielding means and captures foreign matter and liquid;
The liquid discharge suction means and the shielding means are configured to be movable with respect to the container, and the liquid discharge suction means are configured to be relatively movable with respect to the shielding means,
A control unit for controlling the discharge operation and suction operation of the liquid discharge suction means, the movement operation of the liquid discharge suction means and the shielding means, and the driving of the pressure generation means;
The control unit moves the shielding means so as to contact the container and drives the pressure generating means at least during the discharging operation of the liquid discharge / suction means.

  According to the present invention, even when the height of the liquid level in the container fluctuates, it is possible to prevent the liquid in one container from mixing into the liquid in the other container, and foreign matter is present in the liquid in the container. Mixing can be prevented.

(First embodiment)
FIG. 1 shows a schematic diagram of the DNA testing apparatus of this example.

  The DNA testing apparatus 1 includes an extraction unit 2, an amplification unit 3, a hybridization unit 4, a detection unit 5, a control unit 200, and a storage unit 201. The extraction unit 2 extracts DNA from the biological sample, the amplification unit 3 amplifies the DNA, the hybridization unit 4 combines the amplified DNA with the probe DNA, and the detection unit 5 detects whether the DNA is combined with the probe DNA. .

  The control unit 200 controls each operation in the clogging inspection of the filter in addition to the drive control of the liquid discharge suction unit, the pressure generation unit, and the shielding unit, which will be described later. The storage unit 201 stores and holds information necessary for sequence control of the apparatus. In the storage unit 201, for example, the arrangement of the liquid discharge suction means according to the liquid level height, the amount of solution in the container, which will be described later, the discharge, the amount of solution to be sucked, and the suction operation and discharge operation of the liquid discharge suction means Alternatively, operating conditions such as a moving operation and operating conditions of the pressure generating means are stored and held.

  The DNA testing apparatus 1 also accommodates a transport unit (not shown) for transporting a PCR plate (not shown), a pipette unit 6 for supplying and discharging a solution, a pipette transport unit, a pipette tip holding unit, and a used pipette tip. Has pipette disposal section.

  The pipette unit 6 is configured to be movable up and down, front, back, left and right as indicated by arrows 7 by a pipette transport unit using a lead screw, and moves the solution between extraction, amplification and hybridization steps.

  FIG. 2 is a cross-sectional view showing the detailed structure of the pipette unit 6.

  The DNA testing apparatus described in this embodiment can test three samples at the same time, and the pipette unit 6 is configured so that three pipette tips 101 can be attached.

  The pipette tip 101 which is a liquid discharge / suction unit is disposable and discharges and sucks liquid.

  A pipette tip mounting portion 102 that is a liquid operation means is configured such that a cylinder can be moved up and down inside, and a pipette tip 101 is detachably held at the tip. The pipette tip 101 is configured to be movable relative to the duct 103.

  The duct 103 is a hollow circular member formed so as to cover the periphery of the pipette tip 101. That is, the duct 103 shields the inside of the pipette tip 101 and the corresponding container (for example, see FIG. 8) from the inside of the other pipette tips 101 and the container. The material of the duct 103 is not particularly limited, and may be either metal or resin as long as no foreign matter is mixed in from the surroundings.

  The filter member 104 has fine holes, and has a function of capturing and holding foreign matter or liquid while allowing air to pass therethrough. The filter member 104 is attached to an opening formed on the side wall surface of the duct 103 and is disposed in the vicinity of the end portion on the pipette tip 101 side.

  Referring to FIG. 2, an example in which the filter member 104 is disposed behind the pipette tip 101 is illustrated, but the filter member 104 may be disposed on the near side. Moreover, you may arrange | position both in the back and near side of the pipette tip 101. FIG. The filter member 104 is configured with a pore diameter and density that perform the function of capturing and holding dust and scattered nucleic acids while allowing air to pass through. As long as it fulfills this function, the material and structure are not limited and may be selected from foams, bundles of fibers, membranes, and the like.

  Note that the position of the filter member 104 is not limited to the vicinity of the end as shown in FIG. 2, but is arranged in a position (height), area, and number where air can be effectively taken in and out during the splash treatment described later. May be.

  The elastic body 105 is fixed to the tip of the duct 103 so that when the duct 103 is at the lowest point, the elastic body 105 is in close contact with a container (not shown) without a gap so that no air communication path other than the filter member 104 is formed. ing.

  FIG. 2 shows main members constituting the pipette section 6 of FIG. Reference numeral 131 denotes a base plate of the pipette unit 6.

  Reference numeral 132 denotes a support plate fixed to the base plate 131, to which a motor 133 is fixed, and the motor 133 drives a lead screw 135 via a timing belt 134. Reference numeral 136 denotes a pipette holder to which three pipette tip mounting portions 102 are connected, and is driven up and down by the rotation of the lead screw 135.

  A piston motor holding plate 137 is fixed to the pipette holding jig 136, and the motor 138 is fixed. The motor 138 drives the lead screw 140 via the timing belt 139. 141 is a piston shaft having a piston that moves up and down in the pipette mounting portion 102 at the tip, and 142 is a piston shaft holder to which three piston shafts 141 are connected, and is driven up and down by the rotation of the lead screw 140. .

  The pipette tip 101, the pipette tip mounting portion 102, and the duct 103 are integrally movable in the vertical direction with respect to the base plate 131 by a driving means (not shown). Further, the pipette tip 101 and the pipette tip mounting portion 102 are configured to be movable up and down relatively with respect to the duct 103 by the lead screw 135.

  Reference numeral 143 denotes a seal member made of a rubber material, which is held on the inner wall of the duct 103 and seals the gap when the pipette tip mounting portion 102 moves up and down with respect to the duct 103 so that air does not pass through. ing.

  Reference numeral 107 denotes an absorbing member that is disposed inside the inner wall of the duct 103 and that absorbs and holds a mist-like solution through which air passes and is the same as the filter member 104.

  A flow path 144 extends upward from the duct 103 via the absorbing member 107 and is connected to the buffer space 127, and the fan 106 serving as a pressure generating unit is disposed on the buffer space 127. Thereby, the inside of the duct 103 and the fan 106 are in communication. The fan 106 can apply a positive pressure or a negative pressure in the duct 103. The mist-like solution is moved around the tip of the pipette tip 101 by the pressure generated by the fan 106. The three ducts 103 pass through the respective absorbing members 107 and the flow paths 144, and the three flow paths 144 are assembled in the buffer space 127.

  The three specimens handled in the present embodiment need to handle each solution individually, and in order to ensure the accuracy of the test data, it is absolutely necessary to avoid that solutions containing nucleic acids are mixed between different specimens. Don't be.

  A duct 103 is provided for each pipette tip 101. The sealed space formed between the pipette tip 101 and the container excluding the filter member 104 when the pipette tip 101 is at the lowest point is separated for each pipette tip 101 by the duct 103 so that different samples are not mixed. .

  Further, when the distance between adjacent samples in the container is short and there is a possibility that the air flow may interfere between adjacent samples when the fan 106 is driven, the filter member 104 does not face between adjacent ducts as shown in FIG. What is necessary is just to arrange | position to positional relationship.

  Or the guide member extended in the direction which shields between adjacent containers toward the exterior from the periphery of the filter member 104 should just be formed, and the mutual influence of the airflow between adjacent ducts should be reduced significantly. FIG. 3A is a front view showing a configuration including a guide member, and FIG. 3B is a side view thereof. In the figure, 126 is a guide member that reduces airflow interference between adjacent ducts formed so as to cover the periphery of the filter member 104.

  FIG. 4 shows the PCR plate 8. It is a commercial product made of plastic, and 8 × 12 sample tubes are configured at a pitch of 9 mm. The tip (bottom) portion 9 of each well has a shape that can be fitted to a metal block described later. The well opening 10 may be in an open state, or if it is desired to prevent foreign matter from entering from outside, a film material such as aluminum is adhered and sealed, and before filling the sample tube with the solution, the film portion is formed by a punching means. You may break.

  FIG. 5 is a schematic diagram showing the configuration of the amplifying unit 3.

  The metal block 11 into which the PCR plate is fitted is made of a metal having good thermal conductivity such as aluminum or copper alloy, and 96 wells (holes) are formed as indicated by broken lines.

  A Peltier element 12 for heating the metal block 11 is disposed below the metal block 11, and is coated with grease (not shown) for bringing the contact surface into close contact with each other and reliably transferring heat. There is also a sheet material having a good thermal conductivity as a material that obtains the same effect other than grease.

  Below the Peltier element 12, there is a base plate 13 on which the components of the amplifying unit are arranged. The base plate 13 has a hollow structure with openings 14 and 15, and is not shown connected to the openings 14 and 15. The cooling water flows through the pipe. When the temperature of the metal block 11 is lowered, the cooling of the lower surface of the Peltier element 12 is promoted by this cooling water.

  A heating plate 16 made of a metal having high thermal conductivity such as aluminum or copper alloy is disposed on the PCR plate 8. A Peltier element 17 is disposed on the upper portion of the heating plate 16, and is coated with grease (not shown) for bringing the contact surface into close contact with each other and reliably transferring heat.

  A cooling block 18 formed of a metal material is fixed to the contact surface via grease on the upper part of the Peltier element 17. The cooling block 18 promotes cooling when heat is radiated from the upper surface of the Peltier element 17.

  The cooling block 18 has a hollow structure provided with openings 19 and 20, and has a structure in which cooling water flows through piping (not shown) connected to the openings 19 and 20.

  The heating plate 16, the Peltier element 17, and the cooling block 18 are configured as one unit (top plate unit 21), and the top plate unit 21 is configured to be movable by driving means (not shown).

  When the user sets the PCR plate 8 in the amplification section, the top plate unit 21 is in the retracted position and does not interfere with the setting operation. During amplification, the PCR plate 8 is heated from the upper position in the heating position.

  FIG. 6 is a side view showing a state in which the PCR plate 8 and the metal block 11 are fitted. The PCR plate 8 and the metal block 11 are raised and pressed from below in the drawing by a drive mechanism (not shown) against the heating plate 16 of the top plate unit 21 so that the tip 9 of the PCR plate 8 and the metal block 11 are fitted. It is the structure which a joint part sticks closely.

  Next, the operation of the DNA testing apparatus of this example will be described.

  First, containers filled with reagents are set in the extraction, amplification, and hybrid process units of the apparatus. In the extraction unit 2, biological samples such as blood and urine are set, and a solution containing DNA is obtained by a known means. The PCR plate 8 is set in the amplification unit 3 until the extraction is completed.

  The nucleic acid solution is sucked with the pipette tip 101 (see FIG. 2) attached to the pipette section 6 and moved to the amplification section 3. At this time, the pipette tip 101 is replaced with a new one from the one used in the extraction unit 2.

  The pipette tip 101 is attached by moving the pipette portion 6 to a pipette tip holding portion (not shown). The control unit 200 stops the pipette unit 6 at a position where the pipette tip mounting portion 102 and the pipette tip 101 in the pipette tip holding portion face each other. From here, the pipette part 6 is lowered and the pipette tip 101 is press-fitted. During this time, the fan 106 may be driven to suck ambient air through the duct 103. This allows the ambient air to be clean before the pipette tip 101 is attached. The pipette tip 101 is attached and the fan is stopped after a predetermined time.

  7 and 8 are views showing a state where pipetting is performed on the extraction container.

  When the pipette tip 101 is attached, the pipette unit 6 moves toward the opening of the extraction container containing the nucleic acid solution. In the figure, 108 is an extraction container, and the control unit 200 lowers and stops the elastic member 105 at the tip of the duct 103 having an inner diameter larger than the opening of the extraction container 108 until it contacts the opening of the extraction container 108. FIG. 7 shows a state where the elastic member 105 is stopped by contacting the opening of the extraction container 108.

  In this operation, immediately before the elastic member 105 comes into contact with the extraction container 108, the control unit 200 starts sucking air by driving the fan 106 so that dust and foreign matters in the surrounding air are not pushed into the extraction container 108. To. Since the driving at this time is performed very weakly, the surrounding dust and foreign matter are hardly sucked into the duct 103.

  If the duct 103 can be lowered at such a speed that the air is sufficiently exhausted through the filter member 104 and does not push the air into the extraction container 108, the fan 106 is driven after the duct 103 has been lowered. May be.

  FIG. 8 shows a state in which only the pipette tip 101 is lowered from the state of FIG. 7 to reach a height at which the nucleic acid solution 109 can be sucked and stopped. If the fan has been driven weakly before it has been lowered, the driving of the fan 106 is changed from this point in time to increase the flow rate, thereby reliably processing the mist. Since the elastic member 105 is elastically deformed and is in contact with the opening of the extraction container 108, there is no gap between the contact portion of the duct 103 and the extraction container 108. Therefore, when the control unit 200 drives the fan 106, external air enters the duct 103 as indicated by an arrow 110 via the filter member 104.

  When the solution 109 is sucked, the pipette tip 101 is raised and returned to the inside of the duct 103. The controller 200 continues to drive the fan 106 during this time.

  Since the pipette tip 101 is lowered after the duct 103 is lowered in this way, even if the floating mist in the extraction container 108 is pushed up by the pipette tip 101 and rises, it can be processed without leaking to the outside.

  When the fan 106 is driven, external air is supplied into the duct 103 through the filter member 104. However, dust, foreign matter, and floating nucleic acid that affects the test result existing outside are removed by the filter member 104 and do not enter the duct 103. Accordingly, no dust or foreign matter adheres to the outer peripheral portion of the pipette tip 101. Further, since an updraft is generated in the duct 103, even if splashes occur on the liquid surface when the pipette tip 101 enters the solution 109, the uplift airflow raises the inside of the duct 103, and the absorption member 107 Absorbed. Thereafter, the controller 200 drives and stops the fan for a predetermined time, and then raises the duct 103 and the pipette tip 101 integrally.

  Next, the operation of moving the sucked nucleic acid solution to the container set in the amplification unit will be described.

  First, the control unit 200 integrally moves the pipette tip 101 and the duct 103 to a predetermined position on the amplification container. Thereafter, the control unit 200 stops the duct 103 by lowering the duct 103 until the elastic member 105 at the tip thereof is in contact with the amplification container opening surface. Similar to the extraction container, the amplification container has an opening smaller than the duct 103. Next, the control unit 200 lowers only the pipette tip 101 and stops it when reaching the height at which the nucleic acid solution is discharged.

  The operations of the pipette tip 101, the duct 103, and the fan 106 so far are the same as when sucking from the extraction container. However, since there is a tendency for larger droplets to fly at a higher initial speed than in suction, the processing is performed with a higher flow rate of the fan 106 than in suction. The amplification container is pre-filled with reagents used for the amplification reaction, and the solution in the pipette tip 101 is discharged there.

  When all the solution in the pipette tip 101 is discharged, a stirring operation is performed in order to uniformly mix the reagent in the reaction container and the discharged nucleic acid solution. In the agitation, since droplets tend to float more slowly than in discharge, processing is performed with the fan 106 having a lower flow rate than in discharge. The control unit 200 further lowers only the pipette tip 101 from the discharge height, enters the solution, and moves it to the stirring position. When the stirring position is reached, the controller 200 mixes the solution by repeating suction and discharge of the solution by the pipette tip 101 any number of times. When the stirring is completed, the control unit 200 raises the pipette tip 101 and stores it in the duct 103. During this time, the fan 106 is continuously driven.

  External air is supplied into the duct 103 through the filter member 104 by driving the fan 106. However, since dust and foreign matter from the outside are removed by the filter member 104 and do not enter the duct 103, dust and foreign matter do not adhere to the outer periphery of the pipette tip 101.

In addition, since an ascending airflow is generated in the duct 103 by sucking air, the following splashes rise in the duct 103 and are absorbed by the absorbing member 107.
・ Splashes generated on the liquid surface when the pipette tip 101 enters the solution 109 ・ Splashes generated when the solution discharged from the pipette tip 101 becomes bubbles in the liquid and is broken on the liquid surface. Thereafter, the control unit 200 drives and stops the fan 106 for a predetermined time and then raises the duct 103 and the pipette tip 101 integrally.

  Here, since the duct 103 is sized to cover the container opening and to cover one container, it is blocked from adjacent containers, that is, containers that handle different specimens. Is not mixed in another container.

  The size of the duct 103 of this embodiment is a size that covers the opening of one container, but is not limited to one container, and is not related to contamination, that is, the same specimen is used. You may make it the structure which covers the container group to handle collectively.

  Further, since the duct 103 has a minimum necessary size to cover the container opening, it is not necessary to suck a large amount of air. Therefore, it is not necessary to enlarge the fan which is a pressure generating means, and noise can be suppressed.

  In addition, the pipette tip 101 can move relative to the duct 103. Therefore, the duct 103 can be arranged at an appropriate position in each operation (discharge, suction, stirring) of the pipette tip 101 with respect to the solution. As a result, even if the pipette tip 101 is inserted deeply into the liquid, the duct 103 does not reach the liquid surface and a stable splashing process is possible.

  Next, the amplification process will be described with reference to FIGS.

  After the pipette tip 101 discharges the solution to a predetermined container, the pipette unit 6 is retracted from the PCR plate 8, and the retracted top unit 21 returns to a position facing the opening 10 of the PCR plate 8. The PCR plate 8 and the metal block 11 are raised from the lower side of the drawing with respect to the heating plate 16 of the top plate unit 21 by a drive mechanism (not shown) and pressed to fit the tip 9 of the PCR plate 8 and the metal block 11. The portions come into close contact with each other and the state shown in FIG. At this time, the top plate unit 21 has a structure capable of pressing the container of the PCR plate 8 with a force of 0.49 N to 0.98 N per place, or more in some cases.

  PCR is started in such a state that the container is pressed. DNA in the container is amplified by performing PCR at a predetermined holding time and the number of cycles between about 92 ° C. and 55 ° C. to 72 ° C.

  After completion of amplification, the amplification product in each container is sucked by a pipette and conveyed to a purification unit (not shown).

  Containers similar to extraction containers and PCR plates are also set in the purification section, and the containers are filled with magnetic particles, purification liquid, washing liquid, elution liquid, etc. used for purification in advance. Since the operation of the duct 103 and the pipette tip 101 when discharging the amplification product to the purification container is the same as when the solution is moved from the extraction unit to the amplification unit, description thereof is omitted.

  Purification is performed in a container filled with magnetic particles in the purification container. The magnetic particles are filled in 10 μl. After 50 μl of the purified solution and about 50 μl of the amplification product are discharged and stirred, the pipette tip 101 is pulled up from the container. After the magnetic particles and the nucleic acid are bound, the solution is sucked by the pipette tip 101 in a state where a magnet (not shown) is brought close to the container and the magnetic particles are captured on the wall surface of the container. During this time, the fan 106 is continuously driven. After a predetermined time has elapsed after the pipette tip 101 is lifted, the fan 106 is stopped and the duct 103 and the pipette tip 101 are raised integrally.

  Since the solution sucked here is a waste liquid, it is discharged to a predetermined waste disposal unit. The movement and discharge operation of the duct 103 and the pipette tip 101 to the disposal unit at this time are the same as the movement and discharge of the solution described so far.

  At the time of discharge, it is driven at a larger flow rate than at the time of suction, agitation, entry and withdrawal of the pipette tip 101.

  Next, 150 μl of the cleaning solution is transferred to the container where the magnetic particles remain. The operation of the duct 103 and the pipette tip 101 when moving is the same as before. However, at this time, since almost no liquid remains in the container in which the magnetic particles remain, the liquid is directly discharged onto the wall surface of the container, and the distance over which the bounced liquid scatters may be longer. The same applies to the eluate to be filled next to the cleaning liquid.

  Further, when discharging to the waste liquid section, it is desirable not to insert the pipette tip 101 into the waste liquid in order to avoid the waste liquid from being mixed in the subsequent steps. Therefore, when the process progresses and the waste liquid increases, the liquid level of the waste liquid part rises and the discharge position must be raised, and the occurrence of droplet scattering and rebounding moves to a higher position. FIG. 9 shows the state. 111 is a waste liquid container, 112 is the tip position of the pipette tip 101 when it is first discarded, and then gradually rises to the second 113 and the third 114.

  Therefore, the control unit 200 performs control such that the fan 106 is driven strongly or for a long time when the position where the rebound is expected to be low. When the liquid level is high, it may be driven for a long time or for a long time, but when the liquid level is high, in order to avoid the adverse effect of applying unnecessary strong pressure to the liquid level and splashing droplets, It is desirable to drive appropriately.

  In the present embodiment, the control unit 200 changes the control for the cleaning liquid and the elution liquid that are discharged when the container is almost empty. That is, in this case, the control unit 200 drives the fan 106 strongly, or drives the fan 106 for a long time, or both, and reliably collects the mist generated at the bottom of the container in the duct 103.

The following two ways can be considered for determining the control conditions (driving force and driving time) of the fan 106.
1) The control condition is determined by a combination of the detection result of the liquid level in the container obtained by using known means and the operation condition of the pipette tip 101 in the future. In addition, the following can be applied as a known means for detecting the liquid level in the container. For example, it is conceivable to use the pressure change in the pipette tip 101 when the tip of the pipette tip 101 is in contact with the liquid surface or the change in the resistance value of the conductive pipette tip 101. The pipette operating conditions represent the amount of suction and discharge, time and amount of stirring, and the number of times (time).
2) The operation sequence is monitored, and the control condition is determined by a combination of the liquid level height, the pipette arrangement at that time, and the operation condition stored in advance in the storage unit 201 at each stage. Here, in addition to the liquid level, the storage unit 201 also stores information on the amount of solution in the container, and the amount of solution to be discharged and sucked.

  That is, the method 1) is a method in which the control unit 200 determines the control condition of the fan 106 each time, and the method 2) is based on the information stored in the storage unit 201 and sets the control condition of the fan 106 in advance. It can be said that it is a method to decide.

  In this apparatus, it is possible to clean the inside of the apparatus after the inspection is completed and before the pipette tip 101 is discarded. Since it is desirable to avoid dust and foreign matter from adhering to the pipette mounting portion 102 of the dispenser, the pressure generating means is driven while the pipette tip 101 used for the inspection is mounted on the dispenser, and air is blown into the duct 103. It is a method of taking in.

Basically, when the liquid is operated with the pipette tip 101, the droplet periphery is processed in a substantially sealed state, so that the possibility that the nucleic acid is floating in the air is very low. However, just in case, before the next inspection, air is sucked while stopping or moving at an arbitrary position in the apparatus. Thereby, the possibility of contamination can be further reduced. At this time, the pressure adjusting means may be driven more than the flow rate at the time of discharge.
(Second embodiment)
In the first embodiment, the air inside the duct is sucked by the fan. However, the DNA testing apparatus to which the present invention is applied can pressurize the inside of the duct by the fan. Examples thereof will be described below. Members and the like that perform the same functions as in the first embodiment will be described using the same reference numerals.

  The structure around the duct 103 is almost the same as in the first embodiment. The difference is that an absorbing member 115 capable of holding foreign matter and liquid is disposed in the vicinity of the opening on the side of the extraction container 108 shown in FIG. This may be made of the same material as the absorbing member 107 disposed inside the duct described in the first embodiment. The absorbing member 115 is circular like the duct 103 and is arranged around the entire opening of the extraction container 108 (the same applies to other containers).

  As in the first embodiment, the operation when moving from the extraction unit 2 to the amplification unit 3 will be described.

  The pipette tip 101 is moved toward the opening of the extraction container 108 containing the nucleic acid solution. The duct 103 configured to be larger than the opening of the extraction container 108 is lowered and stopped until the elastic member 105 at the tip thereof contacts the absorbing member 115. The fan 106 is driven to suck air into the duct 103 until it comes into contact after being lowered to some extent, so that dust and foreign matters in the surrounding air are not pushed into the container. Since the driving at this time is performed very weakly, the surrounding dust and foreign matter are hardly sucked into the duct 103.

  In addition, if the duct 103 can be lowered at such a speed that the air is sufficiently exhausted through the filter member 104 disposed in the vicinity of the front end of the duct 103 and the air is not pushed into the extraction container 108, the fan 106 is not necessarily driven. You don't have to.

  As the duct 103 is lowered and stopped, the driving of the fan 106 (not shown) is also stopped. Next, only the pipette tip 101 is lowered, and when it reaches a height at which the nucleic acid solution can be sucked, the lowering of the pipette tip 101 is stopped. At this time, the fan starts to be driven before the tip of the pipette tip 101 enters the solution, and air is supplied from above the duct 103. There is no gap between the contact portion of the duct 103 and the extraction container 108 by the elastic member 105, and air is discharged to the outside via the filter member 104 when the fan is driven.

  In this embodiment, the absorbing member 115 is formed on the container side, but it may be at the tip of the duct 103. At this time, if the absorbing member 115 is made of an elastic material and a material that allows air to pass through, the elastic member 105 and the filter member 104 can be used together.

  Further, even in the case where the absorbing member 115 is arranged near the opening as in the second embodiment, it is natural that a method of sucking air with a fan as in the first embodiment can be configured.

  When the solution is sucked, the pipette tip 101 is raised and returned to the duct 103. During this time, the fan 106 is continuously driven. When the fan 106 is driven, the air inside the duct 103 is discharged to the outside through the filter member 104. However, the solution that has been misted during pipetting flows downward and returns into the extraction container 108 or is held by the filter member 104 or the absorption member 115 and is not discharged outside. Therefore, it is possible to prevent nucleic acids that affect the test result from floating in the apparatus. Thereafter, the fan is driven and stopped for a predetermined time, and then the duct 103 and the pipette tip 101 are integrally raised.

  When performing both suction and pressurization as in this embodiment, it is desirable to have a configuration in which a plurality of flow paths are provided and the suction path and pressurization path are provided independently of each other. FIG. 11 shows a configuration example in which the suction path and the pressure path are formed separately. A buffer space 128 in which the suction fan 116 and the suction flow path 117 are gathered, and a buffer space 129 in which the pressurization fan 118 and the pressurization flow path 119 are gathered are individually configured. The control unit 200 drives only the necessary one of the suction fan 116 or the pressurizing flow path 119. An absorption member 145 for suction and a filter member 146 for pressurization are disposed on the inner wall of the duct 103.

  In order to prevent the suction filter member 145 and the pressurization filter member 146 from affecting the mutual suction pressure at all, a configuration using a valve may be used.

  FIG. 12 shows a configuration example provided with a valve.

A valve 147 is provided on the suction member 145 opposite to the side on which the suction fan 116 is disposed. Further, a valve 148 is disposed on the side of the pressurizing filter member 146 opposite to the side on which the pressurizing fan 118 is disposed.
It is the composition which arranged.

  When the controller 200 drives the suction fan 116 to process mist or the like from the duct 103 via the suction flow path 117, the control unit 200 opens the valve 147 and keeps the valve 148 closed.

  On the other hand, when the controller 200 drives the pressurizing fan 118 to process mist or the like from the duct 103 via the pressurizing flow path 119, the control unit 200 opens the valve 148 and keeps the valve 147 closed.

  By performing such control by the control unit 200, no negative pressure is applied to the pressurizing filter member 146 at the time of suction. Therefore, the nucleic acid or the like once held on the pressurizing filter member 146 is peeled off and re-entered in the flow path. There is no floating. Further, since one valve is closed, there is no inflow of air from the pressurizing flow path at the time of suction and from the suction flow path at the time of pressurization, and the drive of the fan can be efficiently transmitted to the container direction.

  11 and 12 show a configuration in which the suction fan 116 and the pressurization fan 118 are individually used, but a common fan may be used and the control unit 200 may change the driving direction each time.

  FIG. 13 shows a configuration example provided with a fan whose driving direction can be changed.

  The configuration is such that the valves 121 and 122 are arranged in the buffer space 130 in front of the fan 120 to open the necessary valve. By adopting such a configuration, it is possible to prevent the mist and foreign matter sucked once and held by the absorbing member from being pressurized and floating in the apparatus again.

  In both the suction system of the first embodiment and the pressurization system of the second embodiment, a pressure sensor may be arranged in the flow path so that filter clogging can be detected. This will be described below.

  In FIG. 14, reference numeral 123 denotes a flat plate arranged in a clogging detection unit configured at an arbitrary position in the apparatus. Reference numeral 124 denotes a pressure sensor arranged in the flow path to detect the pressure in the duct 103 while the fan is driven. A fan driving control unit 125 is connected to the pressure sensor 124 and receives a signal to control driving of the fan. Note that the fan drive control unit 125 may be included in the control unit 200.

  The clogging detection is performed at a timing other than during the inspection at a predetermined period, for example, every day, every week, or every month according to the procedure of the operation flow shown in FIG.

  The pipette unit 6 is moved on the flat plate 123 every predetermined period and lowered until the elastic member 105 at the tip of the duct 103 contacts the flat plate 123 (step S1).

  After being lowered to a predetermined position and stopped, a fan (not shown) is driven to perform suction or pressurization at a flow rate in normal operation for a required time. The pressure at this time is monitored by the pressure sensor 124 (step S2).

  If the pressure value within the predetermined time (differential pressure from the atmospheric pressure) is within the predetermined range, the control unit 200 determines that there is no problem (Y) (step S3) and ends the clogging inspection (step S4). ). In this case, the filter is not clogged, and it is possible to continue the inspection by this apparatus.

  On the other hand, in step S3, when the differential pressure from the atmospheric pressure increases with time and becomes a predetermined value or more (N), the fan driving is stopped at that time, and the duct 103 is once raised to raise the pressure in the duct 103. Return pressure to atmospheric pressure. The duct 103 is again lowered to a predetermined position or lower, the flow velocity is lowered from the previous time, and the suction or pressurization is performed for a longer time (step S5). If the difference from the atmospheric pressure at this time is within a predetermined range, the control unit 200 determines that there is no problem (Y) (step S6), ends clogging detection, and returns to the standby state.

  The fan drive control in the subsequent inspections is carried out by changing to the drive conditions at this time. At this time, the control unit 200 notifies the display unit (not shown) of the apparatus that the filter replacement time is approaching (step S7), and ends the clogging detection (step S8). In this case, the filter is not clogged, and it is possible to continue the inspection by this apparatus.

  In step S6, when the pressure does not fall within the predetermined range within the predetermined time, the control unit 200 repeatedly confirms the pressure by decreasing the flow rate several times (steps S9 to S11).

  If the increase in the differential pressure from the atmospheric pressure does not stop even when driven at a reduced flow rate to a predetermined value, the control unit 200 stops driving the fan at that time. And the control part 200 alert | reports to a display part that a filter is replacement | exchange time (step S12), and does not perform a subsequent test | inspection (step S13).

  In addition, a time until the pressure difference from the atmospheric pressure in the duct 103 when the fan is driven becomes equal to or higher than a predetermined value (when not exceeded, the driving is stopped at a certain upper limit value), and a predetermined time in normal operation, There is also a method of calculating a time difference by comparison and performing control based on the difference. In other words, when the time difference of the comparison results falls within a predetermined range, it is informed that the filter replacement time is near, and when the predetermined time in normal operation becomes longer, the filter is in time for replacement. However, the subsequent inspection should not be performed.

  The pressure sensor 124 may be arranged for each duct 103 as shown in FIG. 14, or one pressure sensor 124 may be arranged in a portion where the ducts 103 are assembled and assembled. In the former case, if one of the plurality of ducts 103 detects clogging, the subsequent inspection may not be performed, or the clogged portion is informed but only the remaining portion is used for inspection. It may be a continuing specification.

  Moreover, the duct part front-end | tip with which the filter part was comprised can be attached or detached, and a user may replace | exchange the duct with a filter, and the service person may replace | exchange.

  As described above, by appropriately performing the filter clogging inspection and the filter replacement, it can be used without causing any adverse effect such as inadvertently applying a large pressure to the liquid surface and causing droplets to scatter around. .

It is a schematic perspective view of the DNA test | inspection apparatus in 1st Example of this invention. It is a schematic block diagram of the pipette part in 1st Example of this invention. It is the front view and side view of a structural example of the duct provided with the guide member for filters. It is a perspective view of an example of a PCR plate applicable to the present invention. It is a typical side view which shows the structure of the amplification part in 1st Example of this invention. It is a side view which shows the state in the amplification reaction of the amplification part shown in FIG. In 1st Example of this invention, it is a side view which shows the state which the duct contact | connected the container. In 1st Example of this invention, it is a side view which shows the state which is attracting | sucking air in a duct. In 1st Example of this invention, it is a figure which shows the position of the pipette tip front end according to discharge in a waste liquid part. It is a side view of the pipette part in 2nd Example of this invention. In 2nd Example of this invention, it is the schematic of the pipette part of the structure which divided | segmented the suction path | route and the pressurization path | route. In 2nd Example of this invention, it is the schematic of the pipette part of the other structure which divided | segmented the suction path | route and the pressurization path | route. In 2nd Example of this invention, it is the schematic of the pipette part of another structure which divided | segmented the suction path | route and the pressurization path | route. It is a figure which shows the condition of the clogging test | inspection of the filter in the DNA test | inspection apparatus of this invention. It is a flowchart of the operation | movement flow in the clogging inspection of a filter.

Explanation of symbols

101 Pipette Tip 103 Duct 104 Filter Member 106 Fan 107 Absorber 200 Control Unit

Claims (13)

A plurality of liquid discharge suction means for discharging the liquid into the container and sucking the liquid held in the container;
A plurality of shielding means for shielding the interior of the container corresponding to the liquid ejection and suction means and the liquid ejection and suction means from the interior of the other liquid ejection and suction means and the other containers;
A pressure generating means that communicates with the shielding means and applies pressure to the shielding means;
An absorbing member that is disposed inside the shielding means and captures the foreign matter and the liquid;
The liquid discharge suction means and the shielding means are configured to be movable with respect to the container, and the liquid discharge suction means are configured to be relatively movable with respect to the shielding means,
A control unit for controlling the discharge operation and suction operation of the liquid discharge suction unit, the movement operation of the liquid discharge suction unit and the shielding unit, and the driving of the pressure generation unit;
The control unit is a liquid inspection apparatus that moves the shielding means to contact the container and drives the pressure generating means at least during the discharging operation of the liquid discharging and sucking means. Provided on the wall surface of the shielding means in communication with the outside, and has a filter that allows air to pass therethrough and traps foreign matter and the liquid;
The space formed by the shielding means and the container in a state in which the shielding means is in contact with the container communicates with the outside only through the filter and the pressure generating means. Liquid testing equipment.   The liquid inspection apparatus according to claim 2, wherein the filter is disposed at a position not facing the container corresponding to the adjacent shielding unit. Pressure detecting means for detecting the pressure in the shielding means, and notification means for notifying the replacement time of the filter,
When the control unit determines that the differential pressure between the pressure detected by the pressure detection unit and the atmospheric pressure when the pressure generation unit is driven for a predetermined time is greater than or equal to a predetermined value, the control unit 4. The liquid according to claim 2, wherein the driving condition of the pressure generating unit is repeatedly changed until the differential pressure becomes equal to or less than the predetermined value, and the notification unit is notified that the replacement time of the filter is approaching. Inspection device.   When the control unit determines that the differential pressure is equal to or greater than a predetermined value, and the differential pressure does not fall below a predetermined value even when the control unit repeatedly changes the driving condition of the pressure generating unit, The liquid inspection apparatus according to claim 4, wherein the control unit stops driving the pressure generating unit and informs the notification unit that the filter member must be replaced. Pressure detecting means for detecting the pressure in the shielding means, and notification means for notifying the replacement time of the filter,
The control unit compares the time from when the pressure generating unit is driven until the differential pressure between the pressure detected by the pressure detecting unit and the atmospheric pressure becomes a predetermined value or more, and a predetermined time in normal operation. 4. The liquid inspection apparatus according to claim 2, wherein a time difference is calculated, and when the time difference is within a predetermined range, the control unit causes the notification unit to notify that the time for replacement of the filter is approaching. .   As a result of the comparison, when the predetermined time in the normal operation is longer than the time until the predetermined value or more, the control unit stops the driving of the pressure generating means, and informs the notification means to the filter member The liquid inspection apparatus according to claim 6, which informs that it has to be replaced.   During the suction operation of the liquid discharge suction means, during the stirring operation of the liquid in the container by repeating the discharge operation and the suction operation of the liquid discharge suction means, and for the liquid in the container The liquid inspection apparatus according to claim 1, wherein the controller drives the pressure generation unit when the liquid discharge / suction unit enters or exits.   9. The liquid inspection apparatus according to claim 1, wherein the control unit drives the pressure generating unit until the shielding unit is moved and brought into contact with the container. A liquid level detector for detecting the liquid level of the liquid in the container;
The control unit determines a driving condition of the pressure generating unit based on a detection result by the liquid level detecting unit and an operating condition of the liquid discharge / suction unit, and drives the pressure generating unit based on the driving condition. The liquid inspection apparatus according to any one of claims 1 to 9. Storage means for storing information on the amount of liquid in the container and the arrangement and operating conditions of the liquid ejection and suction means in each step for inspecting the liquid;
The liquid inspection apparatus according to claim 1, wherein the control unit drives the pressure generating unit based on the information stored in the storage unit. A plurality of communication passages connecting the shielding means and the pressure generating means;
The communication path used for applying a positive pressure in the shielding means by the pressure generating means and the communication path used for applying a negative pressure in the shielding means by the pressure generating means are mutually The liquid inspection apparatus according to claim 1, wherein the liquid inspection apparatus is provided independently.   The control unit moves the shielding unit and the liquid discharge / suction unit to a position away from the container with the liquid discharge / suction unit used for the liquid inspection being mounted after the liquid test is completed, and The liquid inspection apparatus according to claim 1, wherein the pressure generating unit is driven so that a negative pressure is applied to the shielding unit at the distant position.

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