JP2009085797A - Testing method and device using biochip - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a testing method and a device by which the contact area of a sample solution and the wall of a biochip is kept small and a reaction rate between a substance to be tested in a sample solution and a specific binding substance in the biochip is enhanced. <P>SOLUTION: In the biochip 10, the specific binding substance for specifically reacting with the substance to be tested in the sample solution is fixed in a specific sector 16 in an upper face of a plate 12. The sample solution is dripped into the specific sector 16 of the biochip 10, and the substance to be tested in the sample solution is detected by the specific binding substance. The testing device includes a hydrophobic face 18 for keeping the contact face between the droplet 36 of the sample solution and the surface of the plate 12 in the specific sector 16, and a shaking device 34 for changing the shape of the droplet 36 of the sample solution which is dripped into the specific sector 16. <P>COPYRIGHT: (C)2009,JPO&INPIT

Description

  The present invention relates to an inspection method and apparatus using a biochip, and more particularly to an inspection method and apparatus for inspecting a biological substance such as DNA using a biochip.

  The examination of biological substances for examining the presence and amount of in-vivo substances is important for knowing the health condition and determining the treatment method. In addition, in recent years, the inspection of biological substances has been required to be quicker and more sensitive. In particular, the inspection of in-vivo substances that inform health abnormalities is performed in order to expedite treatment and reduce the burden on patients. There is a need for higher sensitivity.

  In the inspection of such a biological substance, there is an inspection method in which only the substance to be extracted is specifically reacted and adsorbed on a specific region of the biochip from the sample liquid extracted from the living body. Since this inspection method can prevent unreacted substances and contaminants from becoming noise and inhibiting detection, the inspection can be performed with high sensitivity.

  By the way, in the inspection of biological substances, the amount of liquid used for the inspection is very small in order to reduce the burden on the patient due to sample collection or to reduce the cost when the chemical liquid is scarce and expensive. There are many cases. When the amount of liquid is very small, the representative dimension of the liquid is also small, so the ratio of the surface area to the volume is large, and the ratio of the contact area of the liquid that comes into contact with the container wall surface when the liquid is placed in the container is large. As a result, there is a possibility that the detection target substance in the liquid (hereinafter also referred to as a test substance) is adsorbed non-specifically outside the specific region of the wall surface.

  When a test substance is adsorbed outside a specific area, the problem of reduced test sensitivity occurs, and when quantification is required for the test, the test substance is adsorbed at an unintended location, resulting in measurement accuracy. Problem arises.

  In addition, when the amount of the liquid is very small, the contact area ratio between the liquid and the wall surface increases, so that the viscous force becomes dominant and the flow in the liquid is suppressed. As a result, since the amount of the test substance supplied to the wall surface decreases, the reaction speed decreases accordingly, and there arises a problem that the inspection time becomes long.

  With respect to the problem of a decrease in the reaction rate, an inspection apparatus with an improved substance supply rate has been developed. For example, a DNA chip reaction promoting device in which a container is formed into a thin film and a part thereof is formed of an elastic body has been commercialized. According to this DNA chip reaction promoting device, a part of the container is elastically deformed to generate a flow in the liquid, so that the reaction rate can be improved.

On the other hand, Patent Document 1 describes an inspection apparatus that mixes liquid by supplying liquid between two parallel wall surfaces and moving one of the wall surfaces relative to the other wall surface. According to this inspection apparatus, since the interface between the liquid layer and the medium functions as an elastic film, the mixing speed of the liquid can be improved.
JP-T-2002-509247

  However, all of the inspection apparatuses described above are for improving the reaction rate, and the problem that the contact area between the liquid and the wall surface is large cannot be solved. For example, the above-described DNA chip reaction promoting device is configured to generate a liquid flow in a state where the liquid is in contact with the entire wall surface, and the reaction rate cannot be improved when the contact area is reduced. On the other hand, the prior art document 1 has a configuration in which the liquid mixing rate increases as the contact area increases, and the reaction rate cannot be improved in a state where the contact area is reduced. Therefore, in the conventional apparatus, there is a problem that the contact area between the liquid and the wall surface is kept small and the reaction speed in the liquid cannot be improved, and both the inspection accuracy and the inspection time cannot be sufficiently improved. there were.

  The present invention has been made in view of such circumstances, and keeps the contact area between the sample liquid and the substrate wall surface of the biochip small, and the reaction between the test substance of the sample liquid and the specific binding substance of the biochip. An object of the present invention is to provide an inspection method and apparatus using a biochip capable of increasing the speed and sufficiently improving the inspection accuracy and the inspection time.

  In order to achieve the above object, the invention according to claim 1 uses a biochip in which a specific binding substance that specifically reacts with a test substance in a sample solution is immobilized in a specific region of a substrate wall surface, In the inspection method of detecting a test substance in the sample liquid by the specific binding substance by supplying a liquid to the specific area, a contact surface between the droplet of the sample liquid and the substrate wall surface is in the specific area. The reaction between the test substance and the specific binding substance is promoted by deforming the droplet of the sample liquid while keeping it.

  According to the present invention, the liquid droplet is deformed while keeping the contact surface of the liquid droplet of the sample liquid in the specific region, thereby causing a flow inside the liquid droplet, and the test substance and the specific binding substance Contact frequency increases and specific reaction is promoted. Thereby, inspection time can be shortened. In addition, according to the present invention, since the contact surface of the liquid droplet is kept in the specific area, it is possible to prevent the liquid droplet from contacting the specific area. Therefore, nonspecific adsorption outside the specific region can be prevented, and the inspection accuracy can be increased. Therefore, according to the present invention, it is possible to achieve both shortening of inspection time and improvement of inspection accuracy. In the present invention, a droplet means having a non-contact portion with a solid around a specific region.

  According to a second aspect of the present invention, in the first aspect of the invention, the movable wall surface is relative to the substrate wall surface in a state in which the movable wall surface is in contact with the sample liquid droplets supplied to the specific region from above. The liquid droplets between the movable wall surface and the substrate wall surface are deformed by moving the liquid crystal to the surface.

  According to the present invention, one of the two wall surfaces in contact with the sample liquid droplet is moved with respect to the other wall surface, so that the droplet can be deformed. In the present invention, the movable wall surface and the substrate wall surface may each be formed of a plate-like plate. Further, in the present invention, the movable wall surface is preferably moved in a direction (that is, up and down direction) to advance and retreat (approach and separate) with respect to the substrate wall surface, but the movable wall surface is moved on the surface (that is, in the horizontal direction). You may do it.

  According to a third aspect of the present invention, in the first aspect of the invention, the movable wall surface is deformed with respect to the substrate wall surface in a state where the movable wall surface is in contact with the sample liquid droplets supplied to the specific region from above. Thus, the liquid droplet between the movable wall surface and the substrate wall surface is deformed.

  According to the present invention, one of the two wall surfaces in contact with the sample liquid droplet is deformed with respect to the other wall surface, so that the droplet can be deformed. In the present invention, the substrate wall surface may be formed of a plate-like plate, and the movable wall surface may be formed of an elastic body or a thin film.

  According to a fourth aspect of the present invention, in the second or third aspect of the invention, the droplet is deformed by applying a repetitive motion having periodicity to the movable wall surface.

  According to the present invention, by giving a repetitive motion to the movable wall surface, the movable wall surface can be moved or deformed, and the droplet can be deformed. In addition, according to the present invention, since the repetitive motion having periodicity is given, the liquid droplets of the sample liquid are less likely to spread out of the specific area than when the non-periodic vibration is given, and the inspection accuracy is improved. It can certainly be improved.

  According to a fifth aspect of the present invention, in any one of the first to fourth aspects of the present invention, a step is formed on the wall surface of the substrate along an outer edge of the specific region, and the droplet of the sample liquid and the substrate are formed by the step. A contact surface with the wall surface is held in the specific region.

  According to the present invention, since the step is provided at the outer edge of the specific region, it is possible to prevent the sample liquid droplet from spreading outside the specific region. The step is preferably a step in which the entire specific region is convex and the periphery thereof is concave. However, only the outer edge of the specific region may protrude in a ring shape.

  The invention according to claim 6 is the invention according to any one of claims 1 to 4, wherein a boundary line of a hydrophobic surface or a hydrophilic surface is formed along an outer edge of the specific region, and the boundary line of the hydrophobic surface or the hydrophilic surface is formed. By this, the contact surface between the sample liquid droplet and the substrate wall surface is held in the specific region.

  According to the present invention, since the boundary line between the hydrophobic surface and the hydrophilic surface is formed along the outer edge of the specific region, it is possible to prevent the sample liquid droplet from spreading outside the specific region. When the sample liquid is hydrophilic, a ring-shaped hydrophobic surface is formed outside the specific region with respect to the hydrophilic substrate wall surface, or the entire inside of the specific region with respect to the hydrophobic substrate wall surface. Is preferably a hydrophilic surface.

  A seventh aspect of the present invention provides the method according to any one of the first to sixth aspects, wherein after the reaction between the test substance and the specific binding substance, a cleaning liquid is supplied onto the wall surface of the substrate. It is characterized by washing away. According to the present invention, since the droplets are washed away after the reaction, the unreacted liquid can be removed and the inspection can be performed accurately.

  The invention according to claim 8 is the invention according to claim 7, characterized in that the cleaning liquid is supplied from one end of the substrate wall surface and absorbed by the absorbing member at the other end. According to the present invention, the used cleaning liquid can be immediately absorbed and removed.

  The invention according to claim 9 is the invention according to claim 7 or 8, wherein the cleaning liquid is supplied by blowing air into the container of the cleaning liquid and extruding the cleaning liquid, and the cleaning liquid is supplied after the cleaning liquid is supplied. The cleaning liquid is pushed out from the substrate wall surface by continuously supplying the air onto the substrate wall surface. According to the present invention, the cleaning of the substrate wall surface and the removal of the cleaning liquid can be performed continuously, and the inspection time can be further shortened.

  In order to achieve the above object, the invention according to claim 10 uses a biochip in which a specific binding substance that specifically reacts with a test substance in a sample solution is immobilized in a specific region of a substrate wall surface. In the inspection apparatus for detecting the test substance in the sample liquid by the specific binding substance by supplying the liquid into the specific area, the contact surface between the droplet of the sample liquid and the substrate wall surface is located in the specific area. Contact surface holding means for maintaining the liquid droplets, and droplet deformation means for deforming the droplets of the sample liquid supplied in the specific region.

  According to the present invention, since the droplet of the sample liquid is deformed by the droplet deforming means, flow occurs inside the sample liquid, the contact frequency between the test substance and the specific binding substance is increased, and the specific reaction is promoted. . Thereby, inspection time can be shortened. In addition, according to the present invention, the contact surface is maintained in the specific area by the contact surface holding means, so that it is possible to prevent the droplet of the sample liquid from coming out of the specific area. Therefore, nonspecific adsorption can be prevented and the inspection accuracy can be increased. Therefore, according to the present invention, it is possible to achieve both shortening of inspection time and improvement of inspection accuracy.

  According to an eleventh aspect of the present invention, in the tenth aspect of the present invention, the droplet deformation means includes a movable wall surface that comes into contact with the droplet of the sample liquid supplied to the specific region from above, and the movable wall surface is defined as the substrate wall surface. And a wall surface moving device that moves relative to each other.

  According to the present invention, since the movable wall surface is moved by the wall surface moving device, it is possible to deform the droplet contacting the movable wall surface and the substrate wall surface. In the present invention, the movable wall surface and the substrate wall surface may each be formed of a plate-like plate.

  According to a twelfth aspect of the present invention, in the tenth aspect of the invention, the droplet deforming means includes a movable wall surface that comes into contact with the droplet of the sample liquid supplied to the specific region from above, and the movable wall surface is defined as the substrate wall surface. And a wall surface deformation device for deforming.

  According to the present invention, by deforming the movable wall surface by the wall surface deforming device, it is possible to deform the liquid droplet that has contacted the movable wall surface and the substrate wall surface. In the present invention, the substrate wall surface may be formed of a plate-like plate, and the movable wall surface may be formed of an elastic body or a thin film.

  A thirteenth aspect of the present invention is characterized in that, in the eleventh or twelfth aspect of the present invention, the droplet deformation means includes a repetitive motion device that applies a cyclic repetitive motion to the movable wall surface. According to the present invention, a repetitive motion device (for example, a vibration device) can apply a repetitive motion to a movable wall surface, and the movable wall surface can be moved or deformed. In addition, according to the present invention, since the repetitive motion having periodicity is given, the liquid droplets of the sample liquid are less likely to spread outside the specific region than when the non-periodic repetitive motion is given, and the inspection accuracy is ensured. Can be improved.

  The invention according to claim 14 is the invention according to any one of claims 10 to 13, wherein the contact surface holding means is a step formed on the substrate wall surface along an outer edge of the specific region. And According to the present invention, since the step is provided at the outer edge of the specific region, it is possible to prevent the sample liquid droplet from spreading outside the specific region. The step is preferably a step in which the entire specific region is convex and the periphery thereof is concave. However, only the outer edge of the specific region may protrude in a ring shape.

  According to a fifteenth aspect of the present invention, in the invention according to any one of the tenth to thirteenth aspects, the contact surface holding means includes a hydrophobic surface or a hydrophilic surface formed on the substrate wall surface along an outer edge of the specific region. It is a boundary line. According to the present invention, since the boundary line of the hydrophobic surface or the hydrophilic surface is formed along the outer edge of the specific region, it is possible to prevent the sample liquid droplet from spreading outside the specific region. When the sample liquid is hydrophilic, a ring-shaped hydrophobic surface is formed outside the specific region with respect to the hydrophilic substrate wall surface, or the entire inside of the specific region with respect to the hydrophobic substrate wall surface. Is preferably a hydrophilic surface.

  A sixteenth aspect of the present invention is characterized in that, in any one of the tenth to fifteenth aspects of the present invention, an end portion of the substrate wall surface is provided with a cleaning liquid supply device for supplying a cleaning liquid onto the substrate wall surface. To do. According to the present invention, since the cleaning liquid is supplied onto the substrate wall surface by the cleaning liquid supply device, the substrate wall surface can be cleaned.

  According to a seventeenth aspect of the present invention, in the sixteenth aspect of the invention, an absorption member that absorbs the cleaning liquid is provided at an end of the substrate wall opposite to the end where the cleaning liquid supply device is provided. And According to the present invention, the used cleaning liquid can be immediately absorbed and removed, and the cleaning liquid can be prevented from contaminating the surroundings.

  According to an eighteenth aspect of the present invention, in the invention of the sixteenth or seventeenth aspect, the cleaning liquid supply device supplies the cleaning liquid onto the substrate wall surface by blowing air into the cleaning liquid container and pushing out the cleaning liquid. The air is supplied onto the substrate wall surface in succession to the supply of the cleaning liquid. According to the present invention, it is possible to perform liquid extrusion by supplying air after supplying the cleaning liquid, and it is possible to continuously clean the substrate wall surface and remove the cleaning liquid.

  According to the present invention, the liquid droplet of the sample liquid is deformed while the contact surface between the liquid droplet of the sample liquid and the wall surface of the substrate is kept in the specific area, so that the liquid droplet contacts the outside of the specific area. In addition to being able to prevent this, it is possible to generate a flow in the droplet to promote a specific reaction, thereby achieving both improvement in inspection accuracy and reduction in inspection time.

  Preferred embodiments of an inspection method and apparatus using a biochip according to the present invention will be described below with reference to the accompanying drawings. The present embodiment is an example in which the present invention is applied to an inspection apparatus for POCT (Point Of Care Testing), but the application of the present invention is not limited to this. The biochip may be a DNA chip or a microchip.

  FIG. 1 is an exploded perspective view showing a biochip 10 used in the first embodiment.

  As shown in these drawings, the biochip 10 is mainly composed of a plate 12 (corresponding to a substrate) and a cover film 14 (corresponding to a movable wall surface). The plate 12 is made of a hard plate such as a white polystyrene plate, and a circular specific region 16 is formed on the upper surface (corresponding to the substrate wall surface) of the plate 12. In the specific region 16, a specific binding substance (not shown) that specifically reacts with the test substance in the sample solution is immobilized. The specific binding substance may be any substance that specifically reacts with the test substance in the sample solution to obtain information on the test substance. The test substance that can be analyzed in the present invention may be any substance that has a specific binding substance that specifically binds to it in the natural world or that can be prepared by chemical means. On the other hand, the specific binding substance is a substance that can specifically bind to the test substance.

  The combination of the test substance and the specific binding substance includes, for example, an antigen and an antibody, certain saccharides and lectins, biotin and avidin, protein A and IgG, a hormone and its receptor, an enzyme and a substrate, a nucleic acid complementary to a nucleic acid, For example. This combination may be reversed. In the most general example, an antigen is a test substance and an antibody is a specific binding substance. The antibody as the specific binding substance may be a polyclonal antibody or a monoclonal antibody. A plurality of types of antibodies may be used. The class of the antibody is not particularly limited, and it may be IgG or IgM, and may be a fragment of an antibody such as Fab, Fab ′, and F (ab ′) 2. In this embodiment, the specific binding substance is immobilized on the plate 12 in order to examine the specific reaction. However, the present invention is not limited to this, and any substance that participates in the reaction with the test substance may be used.

  The method for fixing the specific binding substance to the plate 12 is not particularly limited. For example, the specific binding substance may be fixed by spraying the specific binding substance from a nozzle of an ink jet apparatus. In addition, the specific binding substance may be fixed directly to the plate 12 or indirectly.

  On the other hand, any material can be used as the wall surface (insoluble carrier) of the plate 12 as long as it can stably hold the specific binding substance (substance involved in the reaction). Preferred materials for the wall surface include polystyrene, polycarbonate, polyvinyl toluene, polypropylene, polyethylene, polyvinyl chloride, nylon, polymethacrylate, gelatin, agarose, cellulose, polyethylene terephthalate, etc., polymer materials, glass, ceramics, metals, etc. It is done.

  The shape of the specific region 16 is particularly preferably a circle, but various shapes such as an ellipse and a rectangle are possible. The number of specific regions 16 is not limited to one, and in the case of a DNA chip, a plurality of specific regions 16 may be spaced apart from each other.

  A hydrophobic surface 18 is formed in a ring shape on the upper surface of the plate 12 along the outer edge of the specific region 16. The method of forming the hydrophobic surface 18 is not particularly limited as long as it exhibits water repellency. For example, the hydrophobic surface 18 is formed by applying water-repellent ink. By providing the hydrophobic surface 18, the droplet of the hydrophilic sample liquid dropped in the specific region 16 is prevented from spreading outside the specific region 16. In this embodiment, since the surface of the sample liquid and the plate 12 is hydrophilic, the outside of the specific region 16 is the hydrophobic surface 18. However, when the surface of the sample liquid and the plate 12 is hydrophobic, The outer side of the specific region 16 may be formed as a hydrophilic surface.

  On the other hand, the cover film 14 is formed in a thin film shape having flexibility. The cover film 14 is laminated on the upper surface of the plate 12 and is adhered to the plate 12 by the bonding tape 20. The joining tape 20 is formed in a frame shape, and is configured to affix the four edges on the upper surface of the plate 12 and the four edges on the lower surface of the cover film 14. The joining tape 20 has a predetermined thickness and acts as a spacer between the plate 12 and the cover film 14. As a result, a space of a predetermined size (hereinafter referred to as a chamber 28) is formed between the plate 12 and the cover film 14, and the sample liquid dropped on the specific region 16 is kept in a liquid droplet state without being crushed. be able to. The size of the gap between the plate 12 and the cover film 14 is preferably 10 mm or less, more preferably 5 mm or less, and even more preferably 3 mm or less because it is difficult to form a droplet when it is too large.

  FIG. 2 shows a state in which the biochip 10 is mounted on the inspection apparatus. 3A to 3E are explanatory diagrams for explaining the operation of the inspection apparatus, and each drawing schematically shows a vertical cross-sectional view of the biochip 10.

  As shown in these drawings, a cleaning liquid container 22 and an absorption pad 24 are provided at the end of the biochip 10. The cleaning liquid container 22 is filled with a cleaning liquid 26 and attached to one end of the biochip 10. The absorbent pad 24 is made of a water-absorbing material such as urethane sponge, and is attached to the end opposite to the cleaning liquid container 22. The cleaning liquid container 22 and the absorption pad 24 may be integrated with the biochip 10 so that they can be carried as a unit. Alternatively, the cleaning liquid container 22 and the absorption pad 24 are attached to the inspection apparatus in advance, and the biochip 10 is set at a predetermined position of the inspection apparatus. May be arranged at both ends of the biochip 10.

  The cleaning liquid container 22 and the absorption pad 24 are each in communication with a chamber 28 between the plate 12 and the cover film 14. The communication method is not particularly limited. For example, an opening (not shown) with a backflow prevention valve is formed in the joining tape 20, and the communication is made through this opening. Accordingly, the cleaning liquid container 22 and the absorption pad 24 are communicated with each other through the chamber 28, and the cleaning liquid 26 in the cleaning liquid container 22 can be supplied to the absorption pad 24 through the chamber 28.

  An opening 30 with a backflow prevention mechanism is provided on the upper surface of the cleaning liquid container 22. A nozzle 32 (see FIG. 3B) of an air supply device (not shown) of the inspection device is connected to the opening 30, and air is supplied into the cleaning liquid container 22 by supplying air from the air supply device. Then, the cleaning liquid 26 in the cleaning liquid container 22 is pushed out into the chamber 28 as shown in FIG. Then, as shown in FIG. 3E, air is continuously fed into the chamber 28 even after all of the cleaning liquid in the cleaning liquid container 22 has been pushed out into the chamber 28, so that air is fed into the chamber 28 from above the plate 12. The cleaning solution is removed.

  As shown in FIG. 2, a vibration device 34 is provided in the inspection device. The vibration device 34 is brought into contact with the upper surface of the cover film 14 when the biochip 10 is set in the inspection device. In that case, it is preferable to comprise so that the upper surface of the cover film 14 may be adsorb | sucked. The vibration device 34 includes an eccentric motor and a piston-type vibrator, and is configured to give the cover film 14 a repetitive motion having periodicity (for example, a repetitive motion having a frequency of about 10 Hz and an amplitude of about 0.5 mm). . Thereby, the cover film 14 is given a repetitive motion from the vibration device 34 and is deformed up and down.

  The test apparatus is configured to detect a specific binding substance in which the test substance in the droplet 36 has specifically reacted. Examples of the method include a turbidimetric method, a latex agglutination method, an enzyme immunoassay method, There are fluorescence measurement method, chemiluminescence measurement method, (immuno) chromatography method, etc. According to each method, optical inspection such as reflection or scattering from below or above, inspection by magnetic force, electrochemical inspection is possible It is. Among these, laser, ultraviolet rays, infrared rays, visible light, X-rays and the like can be used for optical inspection.

  Next, a method of using and operating the inspection apparatus configured as described above will be described with reference to FIGS. 3 (A) to 3 (E).

  First, as shown in FIG. 3A, the cover film 14 is turned, and in this state, the sample solution is dropped onto the specific region 16 of the plate 12. Thereby, a droplet 36 of the sample liquid is formed on the specific region 16.

  Next, as shown in FIG. 3 (B), the cover film 14 is covered and attached to the plate 12. Thereby, the droplets 36 on the specific region 16 are brought into contact with the lower surface of the cover film 14 and are in contact with both the cover film 14 and the substrate 12.

  Next, a repetitive motion is given to the cover film 14 by the vibration device 34. As a result, the cover film 14 is deformed as shown in FIGS. 3B and 3C, so that the droplet 36 between the plate 12 and the cover film 14 is deformed so as to expand and contract in the vertical direction. When the liquid droplet 36 is expanded and contracted up and down in this way, the liquid droplet 36 is deformed while maintaining the contact area with the plate 12 (that is, without increasing the contact area), and by the deformation, the inside of the liquid droplet 36 is deformed. In this way, vertical flow occurs. Thereby, the contact frequency of the test substance in the droplet 36 and the specific binding substance on the surface of the specific region 16 can be increased, the reaction rate can be increased, and the test time can be shortened.

  Further, since the hydrophobic surface 18 is formed outside the specific region 16, the liquid droplet 36 on the specific region 16 is reliably prevented from spreading outside the specific region 16. Thereby, it can prevent that a nonspecific reaction generate | occur | produces outside the specific area | region 16, and can improve the precision of a test | inspection.

  After the time necessary for the reaction between the test substance and the specific binding substance has elapsed, air is supplied to the cleaning liquid container 22 as shown in FIG. 3D, and the cleaning liquid in the cleaning liquid container 22 is pushed out onto the plate 12. The pushed cleaning liquid flows through the chamber 28 between the plate 12 and the cover film 14 from the cleaning liquid container 22 side to the absorption pad 24 side and is absorbed by the absorption pad 24. Thereby, unreacted substances and contaminants in the droplets 36 can be removed from the plate 12, and the inspection can be performed accurately. Moreover, reaction time can be controlled by performing a washing process.

  As shown in FIG. 3E, air feeding is continued until all of the cleaning liquid is pushed out from the cleaning liquid container 22. In particular, when optical measurement is performed, light scattering by the remaining cleaning liquid can be prevented.

  After the drying process, the specific binding substance is examined by a predetermined method (for example, optically). Thereby, the detection result of the test substance can be obtained accurately.

  As described above, according to the present embodiment, the cover film 14 is deformed to deform the droplet 36, and a flow is generated inside the droplet 36. Therefore, the test substance in the droplet 36 is identified. The reaction rate with the specific binding substance on the surface of the region 16 can be increased, and the test time can be shortened. Furthermore, since the washing process and the extrusion process are continuously performed after the reaction between the test substance and the specific binding substance, the test time can be further shortened.

  In addition, according to the present embodiment, since the hydrophobic surface 18 is formed outside the specific area 16, it is possible to prevent the liquid droplet 36 from spreading outside the specific area 16 when the liquid droplet 36 is deformed. Further, according to the present embodiment, since the cover film 14 is given a repetitive motion having periodicity, the liquid droplet 36 repeats deformation in the same range, and the liquid droplet 36 is moved to the specific region 16 by the reaction at the time of deformation. Can be prevented from spreading outside. Therefore, according to the present embodiment, it is possible to prevent the droplet 36 from coming into contact with the outside of the specific region 16 to generate a nonspecific reaction, and the inspection accuracy can be improved.

  In addition, according to the present embodiment, the chamber 28 is formed between the plate 12 and the cover film 14, and the cleaning liquid is supplied into the chamber 28 for cleaning, so that the cleaning liquid is scattered outside. It is possible to prevent the device from being contaminated.

  In the above-described embodiment, the cover film 14 having flexibility is covered. However, as shown in FIG. 4, a plate-like cover plate 38 (not flexible) may be provided. In this case, it is preferable that the cover plate 38 is moved up and down by giving a repetitive motion. Thereby, since the liquid droplet 36 is deformed so as to expand and contract vertically, it is possible to generate a flow in the liquid droplet 36 without increasing the contact area and increase the reaction rate. The cover plate 38 may be moved in the horizontal direction as shown in FIG. In this case, the droplet 36 can be deformed without increasing the contact area of the droplet 36 by adjusting the amplitude of the cover plate 38 to an appropriate range.

  In the above-described embodiment, the hydrophobic surface 18 is formed outside the specific region 16 as the contact surface holding unit for the liquid droplet 36. However, the contact surface holding unit is not limited to this, and for example, the specific region 16 A step may be provided on the upper surface of the plate 12 along the outer edge. For example, the plate 40 shown in FIG. 6 has a recess 40 </ b> A formed in the outer peripheral portion of the specific region 16. The recess 40A is formed in a ring shape along the outer edge of the specific region 16, and the entire specific region 16 is a convex portion. In this case, since the droplet 36 is prevented from spreading outside the specific region 16, the contact surface of the droplet 36 can be held in the specific region 16. Thereby, it can prevent that the droplet 36 contacts the exterior of the specific area | region 16, and nonspecific reaction generate | occur | produces, and can improve a test | inspection precision.

  Further, the plate 42 shown in FIG. 7 has a protruding portion 42 </ b> A formed along the outer edge of the specific region 16, and only the outer edge of the specific region 16 protrudes. Also in this case, it is possible to prevent the liquid droplet 36 from spreading outside the specific region 16, and it is possible to prevent the occurrence of non-specific reactions and increase the inspection accuracy.

10 μl of an anti-anti-mouse 1 gG antibody 64 pM solution was dropped on a polyethylene substrate on which 2 ng / mm ^{2 of} anti-mouse 1 gG antibody was physically adsorbed. In order to prevent nonspecific adsorption, protein was also adsorbed on the substrate. For this reason, the contact angle of the antibody solution with respect to the substrate was reduced, and a φ5 mm groove was cut so as to draw a circumference adsorbing the anti-mouse 1 gG antibody. Next, a polyethylene substrate was pressed from above so as to crush the droplets, and the distance between the upper and lower substrates was set to about 2 mm. This example was made by pressing a vibrator on the upper substrate, moving up and down with a frequency of 10 Hz and an amplitude of 0.5 mm, and measuring the change in the amount of reaction over time. Further, as a comparative example, the change over time in the reaction amount was measured in the absence of vibration.

  In the comparative example, the reaction amount required for 60 minutes was obtained in 15 minutes in this example.

Exploded perspective view showing the biochip of this embodiment The perspective view which shows the state which mounted | wore the test | inspection apparatus with the biochip of FIG. Explanatory drawing explaining the effect | action of this invention The figure explaining the biochip from which the structure of a liquid deformation | transformation means differs The figure explaining the biochip from which the moving direction of a movable wall surface differs from FIG. Illustration of biochip with different contact surface holding means Illustration of biochip with different contact surface holding means

Explanation of symbols

  DESCRIPTION OF SYMBOLS 10 ... Biochip, 12 ... Plate, 14 ... Cover film, 16 ... Specific area, 18 ... Hydrophobic surface, 20 ... Bonding tape, 22 ... Cleaning liquid container, 24 ... Absorption pad, 26 ... Cleaning liquid, 28 ... Chamber, 30 ... Opening 32 ... Nozzle 34 ... Excitation device 36 ... Droplet 38 ... Cover plate 40 ... Plate 42 ... Plate

Claims (18)

Using a biochip in which a specific binding substance that specifically reacts with a test substance in the sample liquid is immobilized in a specific area of the substrate wall surface, supplying the sample liquid to the specific area, In a test method for detecting a test substance with the specific binding substance,
The reaction between the analyte and the specific binding substance is promoted by deforming the droplet of the sample liquid while maintaining the contact surface between the droplet of the sample liquid and the wall surface of the substrate within the specific region. An inspection method using a biochip, characterized in that   By moving the movable wall surface relative to the substrate wall surface in a state where the movable wall surface is in contact with the droplet of the sample liquid supplied to the specific region from above, the movable wall surface and the substrate wall surface The inspection method using the biochip according to claim 1, wherein the liquid droplets are deformed.   The liquid between the movable wall surface and the substrate wall surface is obtained by deforming the movable wall surface with respect to the substrate wall surface while the movable wall surface is in contact with the droplet of the sample liquid supplied to the specific region. The test method using the biochip according to claim 1, wherein the droplet is deformed.   The inspection method using a biochip according to claim 2 or 3, wherein the droplet is deformed by applying a repetitive motion having periodicity to the movable wall surface.   2. A step of the substrate wall surface is formed along an outer edge of the specific region, and a contact surface between the sample liquid droplet and the substrate wall surface is held in the specific region by the step. A test method using the biochip according to any one of -4.   A boundary line of a hydrophobic surface or a hydrophilic surface is formed along an outer edge of the specific region, and a contact surface between the droplet of the sample liquid and the substrate wall surface is formed in the specific region by the boundary line of the hydrophobic surface or the hydrophilic surface. The inspection method using the biochip according to any one of claims 1 to 4, wherein   The biochip according to any one of claims 1 to 6, wherein after the reaction between the test substance and the specific binding substance, the droplet is washed away by supplying a cleaning liquid onto the substrate wall surface. Inspection method using   The inspection method using a biochip according to claim 7, wherein the cleaning liquid is supplied from one end portion of the substrate wall surface and is absorbed by an absorption member at the other end portion.   The cleaning liquid is supplied by blowing air into the cleaning liquid container and extruding the cleaning liquid, and after supplying the cleaning liquid, supplying the air onto the substrate wall surface continuously with the cleaning liquid. 9. The inspection method using a biochip according to claim 7, wherein the cleaning liquid is removed from the substrate wall surface. Using a biochip in which a specific binding substance that specifically reacts with a test substance in a sample solution is immobilized in a specific region of a substrate wall surface, and supplying the sample solution into the specific region, In a test apparatus for detecting a test substance with the specific binding substance,
Contact surface holding means for maintaining the contact surface between the sample liquid droplet and the substrate wall surface in the specific region;
An inspection apparatus using a biochip, comprising a droplet deformation means for deforming a droplet of the sample liquid supplied in the specific region. The droplet deformation means includes
A movable wall surface that comes into contact with droplets of the sample liquid supplied to the specific region from above;
A wall surface moving device for moving the movable wall surface relative to the substrate wall surface;
An inspection apparatus using the biochip according to claim 10. The droplet deformation means includes
A movable wall surface that comes into contact with droplets of the sample liquid supplied to the specific region from above;
A wall surface deformation device for deforming the movable wall surface with respect to the substrate wall surface;
An inspection apparatus using the biochip according to claim 10.   The inspection apparatus using a biochip according to claim 11 or 12, wherein the droplet deformation means includes a repetitive motion device that applies a repetitive repetitive motion to the movable wall surface.   The inspection apparatus using a biochip according to claim 10, wherein the contact surface holding means is a step formed on the substrate wall surface along an outer edge of the specific region.   The biochip according to claim 10, wherein the contact surface holding means is a boundary line of a hydrophobic surface or a hydrophilic surface formed on the substrate wall surface along an outer edge of the specific region. Inspection equipment.   The inspection apparatus using a biochip according to claim 10, wherein a cleaning liquid supply device that supplies a cleaning liquid onto the substrate wall surface is provided at an end of the substrate wall surface.   17. The inspection apparatus using a biochip according to claim 16, wherein an absorption member that absorbs the cleaning liquid is provided at an end of the substrate wall opposite to the end provided with the cleaning liquid supply device. .   The cleaning liquid supply device supplies the cleaning liquid onto the substrate wall surface by blowing air into the cleaning liquid container and pushing out the cleaning liquid, and continuously supplying the cleaning liquid to the substrate wall surface. 18. The inspection apparatus using a biochip according to claim 16 or 17, wherein air is supplied to the air.

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