JP2012202807A - Biochemical inspection device - Google Patents

Abstract

PROBLEM TO BE SOLVED: To uniformly develop a specimen in a development layer for development of the specimen in a biochemical analysis method.SOLUTION: In a biochemical analysis method, an analysis chip 12 with a development layer 12b1 for development of a specimen is used to measure the specimen, in which the specimen is supplied to the analysis chip 12 and ultrasonic vibration is applied to the analysis chip 12 with the specimen supplied.

Description

  The present invention relates to a biochemical analysis method and a biochemical analysis apparatus for supplying a sample to an analysis chip for analyzing a sample such as blood and urine to perform biochemical analysis, and more particularly to biochemistry using a development layer for developing a sample. The present invention relates to a method and a biochemical analyzer.

  Conventionally, a specific color component of a dry analytical element or sample that can quantitatively analyze a specific chemical component or formed component contained in this sample simply by spotting and supplying a small droplet of the sample An electrolyte type dry analytical element capable of measuring ion activity of ions has been developed and put into practical use. Biochemical analyzers using these dry analytical elements are suitable for use in medical institutions, laboratories and the like because they can easily and quickly analyze samples.

  In the colorimetric measurement method using a colorimetric type dry analytical element, after a sample is spotted on the dry analytical element, the sample is held at a constant temperature in an incubator for a color reaction (dye generation reaction), and then the sample The dry analytical element is irradiated with measurement irradiation light containing a wavelength selected in advance by a combination of a predetermined biochemical substance and a reagent contained in the dry analytical element, and the optical density is measured. The concentration of the biochemical substance is obtained using a calibration curve representing the correspondence between the optical density obtained in advance and the substance concentration of the predetermined biochemical substance. On the other hand, the potential difference measurement method using an electrolyte type dry analytical element is included in a sample spotted on an electrode pair consisting of two pairs of the same kind of dry ion selective electrodes instead of measuring the above optical density. The activity of specific ions is determined by quantitative analysis with potentiometry using a reference solution.

  In any of the above methods, a liquid sample is accommodated in a sample container (such as a blood collection tube) and set in the apparatus, and a dry analytical element necessary for the measurement is mounted on the apparatus, and the dry analytical element is mounted from the mounting position. While being transported to the landing part and the incubator, the specimen is supplied from the mounting position to the spotting part by the spotting nozzle of the spotting mechanism and spotted to the dry analytical element.

  On the other hand, in order to analyze samples such as blood and urine, microchips having microchannels are being used. In this microchip, a reagent containing a buffer (buffer solution) is injected into the microchannel by a probe, and then a high voltage is applied to the microchannel. By applying this high voltage, each component in the sample in the flow channel is migrated, and the substance to be measured in the sample is separated by the difference in the migration degree. The separated measurement target substance is identified by the reagent and detected by the detection unit of the biochemical analyzer.

  In the dry analytical element as described above, the specimen is uniformly supplied to the reaction layer by passing the development layer for spreading the specimen uniformly while the specimen is supplied to the reagent layer (reaction layer). Uniform measurement results can be obtained over a wide range of the reaction layer. However, there may be cases where the specimen cannot be sufficiently developed at a position away from the spot on the spreading layer, and the amount of specimen that has moved from the spreading layer to the reaction layer is biased, resulting in uneven reaction on the reaction layer. It was.

  In the case of a flow channel analysis chip that causes a specimen to flow down to a flow path, a deployment layer such as a membrane filter that deploys the specimen is arranged in the flow path, and the specimen is allowed to pass through the development layer. In some cases, the sample is supplied to a sensor unit or the like that detects a detection object to which a binding substance that specifically binds to the detection substance provided in the flow path is fixed. When such a sensor unit is provided on the wall surface of the flow path, the amount of the specimen passing through the flow path is made uniform in the cross section of the flow path by passing through the development layer in order to manage reaction measurement conditions. There is a demand to supply the specimen to the sensor unit after that.

  Here, Patent Document 1 discloses a technique for increasing the diffusion rate of blood into a semipermeable membrane. Specifically, in order to increase the diffusion rate of blood diffusing into the semipermeable membrane, blood flowing through the hollow portion of the hollow fiber exists outside the hollow fiber through the hollow fiber wall membrane (semipermeable membrane). A method of applying ultrasonic vibration when developing into a dialysate solution is disclosed.

  In Patent Document 2, in order to fix the reagent to the porous membrane provided in the dry analytical element, one of the reagent layer and the support is made of a porous matrix and the other material is made of a thermoplastic resin. A method is disclosed in which a reagent layer and a support layer are overlapped at the time of producing an analysis element, and a resin component is inserted into the matrix by vibration heat generated by ultrasonic waves and external pressure.

JP-T-2004-529668 JP-A-8-220083

  However, in the method of Patent Document 1, although the speed of diffusing blood to the semipermeable membrane is increased, the diffusion disclosed in Patent Document 1 is performed by osmotic pressure diffusion due to the difference in concentration between blood and dialysate solution or Since diffusion is performed by applying a potential to the semipermeable membrane, it is unclear whether the method of Patent Document 1 can be applied as it is when the specimen is developed on the development layer by the affinity between the specimen and the development layer as in the above analysis chip. is there.

  In addition, the method of Patent Document 2 is used in a dry analytical element for spotting a specimen or a flow-path type analysis chip that causes a specimen to flow down into a flow path and measures the specimen with a sensor unit or the like provided in the flow path. Cannot be applied because the specimen cannot be supplied as a thermoplastic resin.

  In view of the above, an object of the present invention is to provide a biochemical analysis method and a biochemical analysis apparatus in which a specimen is uniformly developed on a development layer included in an analysis chip.

  A biochemical analysis method and apparatus according to the present invention is a biochemical analysis method or apparatus for measuring a sample using an analysis chip having a development layer for developing the sample, and supplying the sample to the analysis chip In addition, ultrasonic vibration is applied to the analysis chip supplied with the specimen.

  The “developing layer” is any material, thickness, as long as the liquid is spread along the gap surface in the spreading layer by the affinity between the gap surface in the spreading layer and the liquid as in the so-called capillary phenomenon. It may be a structure. For example, a membrane filter or a porous film can be suitably applied as the spreading layer. Further, a hydrophilic membrane filter can be suitably applied to develop the specimen as the development layer.

  The period during which the ultrasonic vibration is applied to the analysis chip may be any part or all of the period from when the sample is supplied to when the components contained in the sample are measured, but the sample is supplied to the analysis chip. Preferably, the period starts as early as possible. Moreover, it is preferable that this period is a sufficiently long period in the range which does not extend over the long period which extends analysis time more than necessary. For example, in the case of a dry analytical element, it is conceivable to apply ultrasonic vibration for a period of about several seconds. The ultrasonic vibration may be applied continuously or may be applied intermittently in a plurality of times.

  Further, the supply of the sample in the biochemical analysis method according to the present invention may be performed by spotting the sample on the analysis chip.

    Further, as the “analysis chip”, any analysis chip may be used as long as it has a developing layer for allowing the specimen to pass uniformly. For example, as an analysis chip that supplies a specimen uniformly developed through a development layer to a reagent layer, a dry analytical element including a plurality of layers including a reagent layer corresponding to a plurality of measurement items for the specimen may be used. In addition, an analysis chip of a type that supplies various substances, reagents, or pretreatment agents to a specimen passing through the development layer by including various substances, reagents, or pretreatment agents necessary for analysis in the development layer. Or an analysis chip of a type that includes a development layer and a reagent layer in order from the upstream of the flow path, and a specimen that is uniformly developed in the cross section of the flow path through the development layer is supplied to the reagent layer provided downstream of the flow path May be used.

  In addition, in the biochemical analysis method according to the present invention, the sample is supplied by the analysis chip provided with a microchannel through which the sample is circulated in the channel member, and a measurement unit is provided in a part of the microchannel. In the case where the sample is provided, the sample may be supplied to a development layer provided on the upstream side of the flow channel from the measurement part of the flow channel member of the analysis chip.

  Moreover, it is preferable to apply the ultrasonic vibration in the biochemical analysis method according to the present invention by applying ultrasonic vibration to the analysis chip from a plurality of directions.

  Moreover, it is preferable that the application of the ultrasonic vibration in the biochemical analysis method according to the present invention is performed by sealing a space in contact with the development layer. In addition, the space in contact with the development layer may be sealed with a piezoelectric element for applying ultrasonic vibration, and the space in contact with the development layer may be sealed with a sealing member that covers the space in contact with the development layer.

  According to the present invention as described above, in a biochemical analysis method or apparatus for measuring a sample using an analysis chip provided with a development layer for developing the sample, the analysis chip can be used when the sample is developed in the development layer. Since the sonic vibration is applied, the movement of the specimen is promoted by the vibration energy applied to the specimen, and the specimen can be uniformly and rapidly developed on the development layer, so that analysis with higher accuracy can be performed.

The partial cross section front view which shows schematic structure of the biochemical analyzer by 1st Embodiment FIG. 1 is a plan view of the main part mechanism excluding the spotting mechanism at the element transfer position of the biochemical analyzer of FIG. Sectional front view of the transport path portion of the dry analytical element of FIG. Top view and bottom view of dry analytical element Schematic which expanded a part of spotting part in a 1st embodiment. The elements on larger scale of the spotting part in the modification of 1st Embodiment Schematic side view showing an apparatus according to a second embodiment The schematic perspective view which shows the external appearance of the sensor chip by 2nd Embodiment. The figure which shows the evaluation result of an Example and a comparative example

  Hereinafter, embodiments of the present invention will be described with reference to the drawings. In this embodiment, it is an example of the automatic analyzer by a biochemical analyzer, FIG. 1 is a partial sectional front view which shows the schematic structure of the biochemical analyzer by 1st Embodiment, FIG. 2 is the biochemical analysis of FIG. FIG. 3 is a plan view of the main part mechanism excluding the spotting mechanism at the element transport position of the apparatus, and FIG. 3 shows the main part of the transport path portion of the dry analytical element including the spotting part of the biochemical analyzer of the first embodiment. FIG. 4 is a plan view and a bottom view of the dry analytical element, and FIG. 5 is an enlarged schematic view of a part of the spotting portion in the first embodiment.

  As shown in FIG. 2, the sample tray 2 has a circular shape, a sample container 11 containing a sample, and an element cartridge containing unused dry analysis elements 12 (colorimetric type dry analysis element and electrolyte type dry analysis element). 13. Mount consumables (nozzle tip 14, diluent container 15, mixing cup 16 and reference liquid container 17). The sample container 11 is mounted via a sample adapter 18, and a large number of nozzle chips 14 are stored and mounted in a chip rack 19.

  Based on FIG. 4, the dry analysis element 12 used as an analysis chip in this embodiment will be described. FIG. 4 shows a colorimetric type dry analytical element 12, which is used to measure the degree of coloration of a spotted specimen. The dry analytical element 12 is formed by holding a measuring element 12b having a reagent layer in a rectangular mount portion 12a on the front and back sides made of plastic. In the center of the surface mounting portion 12a shown in FIG. 4A, a spotting hole 12c is opened to expose the surface of the measurement element 12b, and the specimen is spotted on this part. In addition, a photometric hole 12d is opened at the center of the back surface mount portion 12a shown in FIG. 4B, and the back surface of the measurement element 12b is exposed. The coloration degree of this portion is the photometric head of the biochemical analyzer 1. (Not shown). This colorimetric type dry analytical element 12 is provided with a plurality of types having different measurement items depending on the measurement element 12b made of different reagents (coating materials) in the same shape, and the reagent type information (measurement item number, sample) A dot array pattern 12e is recorded in which element information including a seed number and the like and reagent lot information (manufacturing lot number and other unique numbers related to manufacturing) is set and encoded.

  FIG. 5 shows an enlarged schematic view of a part of FIG. As shown in FIG. 5, the measurement element 12b in the dry analytical element 12 includes a spreading layer 12b1 that uniformly spreads the specimen spotted from the spotting hole 12c toward the photometry hole 12d in the reaction layer below, A reflective layer 12b2 for measuring color development in the reaction, a reaction layer 12b3 that is a reagent layer for reacting the specimen, and a transparent support layer 12b4 that supports each layer of the measurement element 12b and transmits inspection light are laminated in this order. In the present embodiment, the mount portion 12a corresponds to a holding member that holds the measurement element 12b composed of each of the layers including the development layer 12b1.

  In this embodiment, a membrane filter made using cellulose acetate is used as the spreading layer. It should be noted that the present invention is not limited to this embodiment, and the development layer included in the analysis chip for developing the specimen is formed on the gap surface in the development layer by the affinity between the gap surface in the development layer and the liquid as in the so-called capillary phenomenon. Any material, thickness, and structure may be used as long as the specimen spreads out by spreading the liquid along.

  For example, as an example of a membrane filter that can be used for the spreading layer 12b1, a fluorine-containing polymer such as cellulose acetate, cellulose nitrate, regenerated cellulose, polyamide, polycarbonate of bisphenol A, polyethylene, polypropylene, polytetrafluoroethylene, or the like is used. The membrane filter that has been made. Moreover, the thickness of the membrane filter in the dry analytical element of the present invention is in the range of about 30 μm to about 300 μm, preferably in the range of about 70 μm to about 200 μm. The porosity (porosity) of the membrane filter is about 25% to about 90%, preferably about 60 to about 90%. The average pore size of the membrane filter is about 0.01 to about 20 μm, preferably about 0.1 to about 10 μm. Various types of these membrane filters are already commercially available from many manufacturers, and can be selected and used as necessary from these commercially available products.

  Examples of the spreading layer include a woven fabric spreading layer (for example, broad) described in JP-A-55-164356 (corresponding US Pat. No. 4,292,272), JP-A-57-66359 (corresponding US Pat. No. 4,783,315), and the like. Flat knitted fabrics such as poplin), knitted fabric spreading layers described in JP-A-60-222769 (corresponding EP Patent 0 162 302A), etc. (eg, tricot knitted fabric, double tricot knitted fabric, Miranese knitted fabric), JP Fibrous microporosity of organic polymer fiber pulp-containing papermaking paper development layer described in JP-A-57-148250, development layer formed by applying a dispersion of a fiber and a hydrophilic polymer described in JP-A-57-125847, etc. Expanded layer: Continuously formed by adhering fine particles such as membrane filter layer (brush polymer layer) and polymer microbeads described in JP-B-53-21677 (corresponding US Pat. No. 3,992,158) in a point contact with a hydrophilic polymer binder Isotropic microporous exhibition with microvoids Layer, isotropic microporosity containing continuous microvoids in which polymer microbeads described in JP-A-55-90859 (corresponding US Pat. No. 4,258,001) are bonded in a point contact with a polymer adhesive that does not swell with water Non-fibrous isotropic microporous development layer such as a layer (three-dimensional lattice-like granular structure layer) development layer; JP-A 61-4959 (corresponding US Pat. No. 5,019,347), JP-A 62-138756, JP A plurality of microporous layers described in JP-A-62-138757, JP-A-62-138758 (corresponding EP Patent 0 226 465A), etc. (for example, two layers of woven fabric or knitted fabric and membrane filter, woven fabric or knitted fabric) An unfolded layer with excellent blood cell separation ability is formed by laminating and bonding membrane filters and woven fabrics or knitted fabrics) with fine discontinuous or island-like (dots in the printing field) adhesive on their surfaces. It is applicable to the present invention.

  The fabric or knitted fabric used for the spreading layer is subjected to physical activation treatment represented by glow discharge treatment or corona discharge treatment described in JP-A-57-66359 on at least one side of the fabric fabric, or specially. Hydrophobic treatment by water washing and degreasing treatment described in JP-A-55-164356, JP-A-57-66359, etc., hydrophilic treatment such as impregnation with a hydrophilic polymer, or by appropriately combining these treatment steps, thereby making the fabric fabric hydrophilic, The adhesive force with the lower layer (side closer to the support) can be increased. Further, as described in JP-A-59-171864, JP-A-60-222769, JP-A-60-222770, etc., a polymer-containing aqueous solution or a polymer-containing water-organic solvent mixed solution is applied from above the spreading layer. The development area or spread of the liquid sample can be controlled.

  The light reflecting layer 12b2 is a light shielding layer or a white particle such as titanium dioxide fine particles and barium sulfate fine particles that have both light shielding properties and light reflecting properties are dispersed almost uniformly in a hydrophilic polymer binder such as gelatin. The thickness of the layer ranges from about 2 μm to about 20 μm.

  In addition, the dry analysis element may have a layer configuration appropriately changed as long as it includes a development layer and a reagent layer in order from the spotting side of the specimen, and a color shielding layer is provided instead of the light reflection layer 12b2. May be. Further, a known adhesive layer made of a hydrophilic polymer may be provided on the reagent layer, the color shielding layer or the light reflecting layer for the purpose of firmly bonding and integrating the spreading layer. The dry thickness of the adhesive layer ranges from about 0.5 μm to about 5 μm. Further, a surfactant may be contained in the reagent layer, the color shielding layer or the light reflecting layer, the adhesive layer, the developing layer, and the like.

  The element information by the dot arrangement pattern 12e is formed by dot printing at the front and rear center portions in the width direction of the mount 12a on the back surface of the dry analytical element 12. Further, in the vicinity of the central photometric hole 12d, a horizontal striped barcode pattern 12f that includes reagent type information and does not include reagent lot information is formed by printing. A measurement item name 12g is printed on the mount 12a on the surface of the dry analytical element 12. The bar code pattern 12f is for use in an analysis apparatus of a model that cannot read the dot array pattern 12e. Moreover, an electrolyte type dry analysis element may be used for the analysis apparatus 1 of the present invention, and element information is given to the dry analysis element by printing the same patterns 12e and 12f as described above.

  The spotting unit 3 is arranged on an extension of the center line of the sample tray 2 and is used for spotting a specimen such as plasma, whole blood, serum, urine, etc. on the transported dry analytical element 12. 6, the sample is applied to the colorimetric dry analysis element 12 and the sample and the reference liquid are applied to the electrolyte type dry analysis element 12. Subsequent to the spotting unit 3, a chip discarding unit 9 in which the nozzle tip 14 is discarded is arranged.

  The first incubator 4 has a circular shape and is disposed at an extended position of the chip discarding unit 9. The first incubator 4 accommodates a colorimetric type dry analytical element 12 and keeps the temperature constant for a predetermined time to perform colorimetric measurement. The second incubator 5 (see FIG. 2) is disposed at an adjacent position on the side of the spotting portion 3, accommodates the electrolyte-type dry analysis element 12, maintains a constant temperature for a predetermined time, and performs a potential difference measurement.

  The element transport mechanism 7 (see FIG. 3), which is not shown in detail, is disposed inside the sample tray 2 and connects the center of the sample tray 2 and the center of the first incubator 4 to the spotted portion. 3 and an element transport for transporting the dry analytical element 12 from the sample tray 2 to the spotting section 3 and further to the first incubator 4 along a linear element transport path R (FIG. 2) passing through the chip discarding section 9 and the chip discarding section 9. A member 71 (conveying bar) is provided. The element conveying member 71 is slidably supported by a guide rod 38, and is reciprocated by a drive mechanism (not shown). The tip end portion is inserted into the guide hole 34a of the vertical plate 34, and slides in the guide hole 34a.

  The transfer mechanism 8 is also installed as the spotting unit 3, and transfers the electrolyte type dry analytical element 12 from the spotting unit 3 to the second incubator 5 in a direction orthogonal to the element transport path R.

  The spotting mechanism 6 is disposed in the upper part, and a spotting nozzle 45 that moves up and down moves on the same straight line as the above-described element transporting path R, spotting the specimen and the reference liquid, and diluting and mixing the specimen with the diluent. . The spotting nozzle 45 has a nozzle tip 14 attached to the tip, and sucks and discharges a sample, a reference liquid, and the like in the nozzle tip 14, and is provided with a syringe means (not shown) for performing the suction and discharge. The nozzle tip 14 is removed by the tip discarding unit 9 and discarded.

  The element discarding mechanism 10 (see FIG. 2) is attached to the first incubator 4 and pushes the colorimetric dry analytical element 12 after measurement to the center of the first incubator 4 to drop and discard. The element transport mechanism 7 can also be discarded. Further, the electrolyte type dry analytical element 12 after being measured by the second incubator 5 is discarded into the disposal hole 69 by the transfer mechanism 8.

  In addition, a blood filtration unit (not shown) that separates plasma from blood is installed in the vicinity of the sample tray 2.

  The mechanism of each part will be specifically described below. First, the sample tray 2 has a disk-shaped rotating disk 21 that is rotationally driven in the forward direction and the reverse direction, and a disk-shaped non-rotating part 22 at the center.

  As shown in FIG. 2, the rotating disk 21 is adjacent to five sample mounting portions 23 of A to E that hold a sample container 11 such as a blood collection tube containing each sample via a sample adapter 18. The five element mounting portions 24 for holding the element cartridges 13 accommodated in a stacked state of the unused dry analysis elements 12 that normally require a plurality of types corresponding to the measurement items of each specimen, and a number of nozzles Two chip mounting portions 25 that hold a chip rack 19 that stores the chips 14 side by side in the holding holes, a diluent mounting portion 26 that holds three diluent containers 15 that store the diluent, a diluent and a sample, And a cup mounting portion 27 for holding a mixing cup 16 (a molded product in which a large number of cup-shaped recesses are arranged) are arranged in an arc shape.

  Further, the non-rotating part 22 is provided with a cylindrical reference liquid mounting part 28 for holding the reference liquid container 17 containing the reference liquid in the movement range of the spotting nozzle 45 on the extension of the element transport path R, The reference liquid mounting section 28 is provided with an evaporation prevention lid 35 (FIG. 1) that opens and closes the opening of the reference liquid container 17.

  The evaporation prevention lid 35 is held by a swing member 37 pivotally supported by the non-rotating portion 22 at the lower end, and is biased in the closing direction. The upper end locking portion 37a of the swinging member 37 can be in contact with the lower end corner portion 42a of the moving frame 42 of the spotting mechanism 6, and the swinging member 37 is moved in the opening direction by the moving frame 42 that has moved close when the reference liquid is sucked. The evaporation prevention lid 35 opens the reference liquid container 17 so that the reference liquid can be sucked by the spotting nozzle 45. In other states, the evaporation prevention lid 35 closes the opening of the reference liquid container 17 to prevent the reference liquid from evaporating and prevents a decrease in measurement accuracy due to a change in concentration.

  The rotating disk 21 has an outer peripheral portion supported by a support roller 31 and a central portion rotatably held on a support shaft (not shown). Further, a timing belt (not shown) is wound around the outer periphery of the rotary disk 21 and is driven to rotate in the forward direction or the reverse direction by a drive motor. The non-rotating part 22 is non-rotatably attached to the support shaft.

  As shown in FIG. 3, the element cartridges 13 are usually stacked with a plurality of unused dry analytical elements 12 stacked from above. When loaded in the element mounting portion 24, the lower end portion is held on the bottom wall 24a of the element mounting portion 24, and the dry analytical element 12 at the lowermost end portion is positioned at the same height as the element transport surface. An opening 13a through which only one dry analytical element 12 can pass is formed on the front side, and an opening 13b through which the element transport member 71 can be inserted is formed on the rear side.

  In addition, a window portion 13 c is provided on the bottom surface of the element cartridge 13, and a bottom wall 24 a of the element mounting portion 24 is provided on the bottom wall 24 a so that the element information given to the lower surface of the dry analytical element 12 can be read from below the element cartridge 13. Are formed respectively.

  A reader 33 that reads element information based on the dot array pattern 12 e of the dry analysis element 12 is installed below the sample tray 2. In the reader 33, the rotating disk 21 is rotated by the operation of the sample tray 2 from the element transport position shown in FIG. 3, and the sample container 11 (sample mounting portion 23) is a spotting nozzle as shown in FIG. When the element cartridge 13 (element mounting portion 24) containing the dry analytical element 12 used for the measurement of the sample is moved to the suction position on the movement path (element transport path R) of 45, the position is below the position where the element cartridge 13 is moved. is set up. That is, in the illustrated case, the reader 33 is installed at the rotation position of the element mounting unit 24 with a phase angle that is shifted from the element transport path R by the phase pitch between the sample mounting unit 23 and the element mounting unit 24. In FIG. 3, the rotary disk 21 is partially cut away to show the reader 33, and in FIG. 3, the reader 33 is shown below the element mounting portion 24 in the element transport path R for convenience.

  The reader 33 is composed of a CCD camera corresponding to the dot recording method. The reading of the element information of the dry analysis element 12 by the reader 33 is performed prior to the sample suction from the corresponding sample container 11 and the transport of the dry analysis element 12. Then, the reagent type information, reagent lot information, etc. can be obtained from the 6-digit or 4-digit number obtained by reading the element information given to the dry analytical element 12, and from the recording pattern, etc. The direction can be recognized. Thereby, a set failure can be detected and a warning can be issued.

  The sample adapter 18 is formed in a cylindrical shape, and the sample container 11 is inserted from above. The sample adapter 18 includes an identification unit (not shown), and information such as the type of sample (processing information) and the type (size) of the sample container 11 is set. The identification sensor 30 (FIG. 2) provided reads the identification, and determines whether or not the sample is diluted, whether or not the plasma is filtered, and calculates the amount of liquid level fluctuation associated with the size of the sample container 11. Processing control is performed accordingly. For the specimen container 11 that requires plasma filtration, the specimen container 11 is inserted into the adapter 18 and a holder equipped with a filtration filter is mounted via a spacer (both not shown).

  The spotting unit 3 and the transfer mechanism 8 include a support base 61 that is long between the sample tray 2 and the first incubator 4 in a direction perpendicular to the element transport path R, and a sliding frame 62 is movable on the support base 61. is set up. A first element presser 63 and a second element presser 64 in which spot wearing openings 63a (FIG. 3) are formed are attached to the sliding frame 62 so as to be movable integrally with each other. The first element holder 63 (the same applies to the second element holder 64) has a recess 63b on the bottom surface facing the support base 61 through which the dry analysis element 12 passes along the element movement path R. The sliding frame 62 is guided at one end by the guide bar 65, the pin 66 is engaged with the long groove 62a at the other end, and the drive gear 67 of the drive motor 68 is engaged with the rack gear 62b. Is done. The support base 61 is provided with a second incubator 5 and a disposal hole 69.

  The spotting part 3 will be described in more detail with reference to FIG. Although omitted in FIG. 3, as shown in FIG. 5, the piezoelectric element 60 that generates ultrasonic vibrations is disposed on the lower surface of the element presser 63, and the lower surface of the piezoelectric element 60 is disposed with the dry analysis element 12 disposed thereon. It is provided in contact with the upper surface 12h of the dry analytical element. Further, the biochemical analyzer 1 includes a piezoelectric element (in this example, a piezoelectric element) 60 as an ultrasonic wave generating means, and a piezoelectric driver (not shown) that applies a driving voltage for exciting the piezoelectric element 60 to the piezoelectric element 60. And a waveform generator (not shown) that generates a waveform signal that defines the waveform of the drive voltage and inputs the waveform signal to the piezo driver. A signal voltage is applied from the piezo driver to the piezoelectric element 60 for a predetermined period simultaneously with the spotting of the sample on the dry analysis element 12. The ultrasonic vibration generated by this is applied from the upper surface 12h of the dry analytical element arranged at the spotting position. In FIG. 1 and FIG. 3, the thickness of the layer is appropriately different from the actual one for the sake of explanation, and the wiring from the piezoelectric element 60 to the piezo driver is omitted. The predetermined period can be arbitrarily set, and is several seconds in the present embodiment.

  Note that “at the time of sample supply” in the present invention is not limited to the above-described embodiment, and means a part or all of an arbitrary period from when the sample is supplied to when the component contained in the sample is measured. However, the period during which the ultrasonic vibration is applied to the analysis chip may be any time as long as the sample is supplied, but preferably the sample is applied to the analysis chip for any period including when the sample is applied to the analysis chip. Preferably, it is an arbitrary period starting at the earliest possible time. For example, in the above embodiment, if ultrasonic vibration can be applied to the analysis chip at the time of supplying the sample, any time after the sample is spotted on the analysis chip and before the measurement of the analysis chip is performed. Ultrasonic vibration may be applied to the analysis chip. Moreover, it is preferable that the period during which the ultrasonic vibration is applied to the analysis chip is a sufficiently long period as long as the analysis time is not extended over a long period of time. For example, in the case of a dry analytical element, it is conceivable to apply ultrasonic vibration for a period of about several seconds. Further, the ultrasonic vibration may be applied continuously or may be applied intermittently in a plurality of times.

  In addition, ultrasonic vibration may be generated by any vibration mode of the piezoelectric element. For example, thickness vibration, thickness shear vibration, or length vibration may be used.

  Further, the vibration frequency can be arbitrarily set within a range that can promote the development of the specimen on the development layer, and a plurality of ultrasonic vibrations having different periods may be applied from the piezoelectric element to the analysis chip.

  The direction in which the vibration is applied to the analysis chip may be any direction, but it is preferable to match the vibration direction of the vibration element with a direction in which it is particularly desired to uniformly develop the specimen on the development layer. . In addition, it is preferable to apply the vibration by making the vibration direction of the thickness longitudinal vibration of the piezoelectric element coincide with the direction in which it is particularly desired to uniformly develop the specimen on the development layer. For example, in the case of an analytical element that deposits a specimen, the development unevenness occurs in a direction perpendicular to the thickness direction of the development layer from the position where the specimen is spotted. It is preferable to apply sonic vibration, and it is more preferable to apply ultrasonic vibration in a plurality of directions. In addition, ultrasonic vibrations may be applied simultaneously from different directions to the dry analytical element, ultrasonic vibrations may be applied alternately from a plurality of different directions, and these may be combined arbitrarily.

  Further, ultrasonic vibration may be applied to the analysis chip from a plurality of directions. For example, the analysis chip may be a dry analysis element such as vertical double-sided 12h, 12k or horizontal double-sided 12i, 12j in FIG. It is preferable to apply from the opposite direction, for example, by holding the vibration part in contact with both sides. This is because the analysis chip can be firmly brought into contact with the vibration part and supported, and ultrasonic vibration can be effectively applied to the entire analysis chip from a plurality of directions. In such a case, the analysis apparatus 1 includes a known moving mechanism that supports the piezoelectric element 60 and can move the piezoelectric element to a position where the piezoelectric element is brought into contact with a desired portion of the analysis chip, and a control unit thereof.

  As shown in FIG. 2, when the first element presser 63 is positioned at the spotting portion 3, the colorimetric type dry analytical element 12 after the spotting is pushed out by the element transport mechanism and the first incubator 4. It is transferred to. On the other hand, when spotting is performed on the electrolyte type dry analytical element 12, the sliding frame 62 is moved, and the dry analytical element 12 after spotting is held on the support base 61 while being held by the first element presser 63. It is transferred to the second incubator 5 so as to slide, and a potential difference measurement is performed. At that time, the second element presser 64 moves to the spotting unit 3 (spotting position), and then the specimen is spotted on the colorimetric dry analytical element 12 and then transported to the first incubator 4. Can be transported. When the measurement in the second incubator 5 is completed, the sliding frame 62 is further moved, and the dry analytical element 12 after the measurement is transferred to the disposal hole 69 to be dropped and discarded.

  When the colorimetric type dry analytical element 12 is transported, the second element presser 64 is moved to the spotting section 3 and only when the electrolyte type dry analytical element 12 is transported. You may make it move 63 to the spotting part 3. FIG.

  The imaging member 33 reads other information in addition to reading the dot array pattern 12e. For this purpose, an additional light source (not shown) is installed. As this additional light source, a light source having a specific wavelength, such as an infrared light source or a deterioration detection light source, is installed according to the detection mode. The reading of spotting information and other information by the information reader 5 will be described later.

  The spotting mechanism 6 (FIG. 1) includes a moving frame 42 held on a horizontal guide rail 41 of a fixed frame 40 so as to be movable in the lateral direction, and two spotting nozzles that can be moved up and down on the moving frame 42. 45 is installed. A vertical guide rail 43 is fixed to the center of the moving frame 42, and two nozzle fixing bases 44 are slidably held on both sides of the vertical guide rail 43. An upper end portion of the spotting nozzle 45 is fixed to the lower portion of the nozzle fixing base 44, and a shaft-like member extending upward is inserted into the drive transmission member 47. A fitting force of the nozzle tip 14 is obtained by a compression spring interposed between the nozzle fixing base 44 and the drive transmission member 47. The nozzle fixing base 44 can move up and down integrally with the drive transmission member 47, and is driven relative to the nozzle fixing base 44 by compression of a compression spring when the nozzle tip 14 is fitted to the tip of the spotting nozzle 45. The transmission member 47 can move downward. The drive transmission member 47 is fixed to a belt 50 stretched around upper and lower pulleys 49 and moves up and down in accordance with the travel of the belt 50 by a motor (not shown). A balance weight 51 is attached to the outer portion of the belt 50 to prevent the descent nozzle 45 from moving down when not driven.

  The moving frame 42 is driven in the horizontal direction by a belt drive mechanism (not shown), and the horizontal movement and the vertical movement are controlled so that the two nozzle fixing bases 44 independently move up and down. Moves horizontally and moves up and down independently. For example, one spotting nozzle 45 is for a specimen, and the other spotting nozzle 45 is for a diluting liquid and a reference liquid.

  Both the spotting nozzles 45 are formed in a rod shape, an air passage extending in the axial direction is provided inside, and a pipette nozzle tip 14 is fitted in a sealed state at the lower end. Each spotting nozzle 45 is connected to an air tube connected to a syringe pump (not shown) and supplied with suction / discharge pressure. Further, the liquid level of the sample or the like can be detected based on the change in the suction pressure.

  The chip discarding unit 9 is provided so as to intersect the transport path R in the vertical direction, and includes an upper member 81 and a lower member 82. The support base 61 in the chip discarding unit 9 is formed with a drop port 83 that is opened in an elliptical shape. The upper member 81 is fixed to the upper surface of the support base 61, an engagement notch 84 is provided immediately above the drop opening 83, and the lower member 82 is cylindrical so as to surround the lower side of the drop opening 83 on the lower face of the support base 61. The nozzle tip 14 that is formed and falls is guided.

  Then, the spotting nozzle 45 to which the nozzle tip 14 is mounted is moved down in the upper member 81 and then moved in the lateral direction, and the upper end of the nozzle tip 14 is engaged with the engagement notch 84. The nozzle tip 14 is extracted by moving the landing nozzle 45 upward, and the detached nozzle tip 14 is dropped and discarded through the drop port 83.

  The first incubator 4 that performs the colorimetric measurement includes an annular rotating member 87 on the outer peripheral portion, and the rotating member 87 rotates with an inclined rotating cylinder 88 fixed to an inner peripheral lower portion supported by a lower bearing 89. It is free. An upper member 90 is disposed on an upper portion of the rotating member 87 so as to be integrally rotatable. The upper surface of the upper member 90 is flat, and a plurality of (13 in the case of FIG. 1) concave portions are formed on the upper surface of the rotating member 87 at predetermined intervals on the circumference, and a slit-like space is formed between the members 87 and 90. An element chamber 91 is formed, and the height of the bottom surface of the element chamber 91 is the same as the height of the transfer surface. Further, the inner hole of the inclined rotating cylinder 88 is formed in the disposal hole 92 of the dry analytical element 12 after the measurement, and the dry analytical element 12 in the element chamber 91 is moved to the center side as it is and dropped and discarded.

  The upper member 90 is provided with a heating means (not shown), and the temperature of the dry analytical element 12 in the element chamber 91 is kept constant at a predetermined temperature. Further, as shown in FIG. 4, the upper member 90 is provided with a pressing member 93 corresponding to the element chamber 91 to press the mount of the dry analysis element 12 from above to prevent the sample from evaporating. A heat retaining cover 94 is disposed on the upper surface of the upper member 90, while the first incubator 4 is entirely covered with a light shielding cover 95. Further, a photometric opening 91a is formed at the center of the bottom surface of each element chamber 91 of the rotating member 87, and the reflection optical density of the dry analytical element 12 by the photometric head 96 disposed at the position shown in FIG. 2 through the opening 91a. Is measured. The first incubator 4 is rotationally driven by a belt mechanism (not shown) and is driven to reciprocate.

  The disposal mechanism 10 includes a disposal bar 101 that moves forward and backward in the element chamber 91 in the center direction from the outer peripheral side. The disposal bar 101 is fixed to a belt 102 whose rear end travels in a horizontal direction, and the measured dry analytical element 12 is pushed out from the element chamber 91 in accordance with the traveling of the belt 102 driven by the drive motor 103 and discarded. To do. A recovery box for recovering the dry analytical element 12 after measurement is disposed below the disposal hole 92.

  Further, in the second incubator 5 for measuring the ion activity, the first element presser 63 of the sliding frame 62 serves as an upper member, and one element chamber is provided between the upper surface of the measurement main body 97 by the concave portion at the bottom. Is formed. The second incubator 5 is provided with a heating means (not shown), and a temperature measuring portion of the dry analytical element 12 for measuring the ion activity is isothermally heated to a predetermined temperature. Further, three pairs of potential measurement probes 98 for measuring the ion activity are provided on the side portion of the measurement main body 97 so as to come into contact with the ion selective electrode of the dry analytical element 12.

  A plasma filtration unit (not shown) is inserted through a holder (not shown) having a filter made of glass fiber inserted into the specimen container 11 (collecting blood vessel) held in the sample tray 2 and attached to the upper end opening. Plasma is separated and sucked from blood, and the filtered plasma is held in the cup at the upper end of the holder.

  The operation of the biochemical analyzer 1 as described above, setting of measurement conditions, and the like are performed by input from an operation panel 55 (not shown) installed in the housing 20 (not shown). The operation panel 55 (interface) performs various instruction operations by pressing with a fingertip such as a display screen, a start key, a stop key, a specimen key, a consumable key, a manual key, an emergency key, a calibration key, a numeric keypad, and a print key. Operation keys are provided. The operation panel 55 is connected to a control system (not shown), and measurement calculation processing based on a control program registered therein is set, automatic measurement operation, manual measurement operation, emergency measurement operation, calibration operation, and printing operation. Are selected and executed, and the analysis result (component concentration) is calculated based on the measurement information based on the analysis information corresponding to the element information. Then, in order to output and record the measurement results, these data are printed by the printer 57 in order to confirm the set values.

  Next, the overall operation of the biochemical analyzer 1 will be described. First, before performing the analysis, in each of the mounting portions 23 to 28 of the sample tray 2, a specimen container 11 containing each specimen, an element cartridge 13 loaded with a dry analytical element 12, a chip rack 19 containing a nozzle chip 14, The mixing cup 16, the diluent container 15 and the reference liquid container 17 are mounted to prepare for measurement.

  Thereafter, the analysis process is started. First, in the case of a specimen that requires plasma filtration, the whole blood in the specimen container 11 is filtered by a blood filtration unit to obtain a plasma component. Next, the element cartridge 13 of the sample to be measured by rotating the rotating disk 21 is stopped at the element take-out position corresponding to the spotting unit 3, and the dry analysis element 12 is taken out from the element cartridge 13 by the element transport mechanism and is spotted. 3 to transport. Note that the analysis information given to the dry analysis element 12 is read before being transported to the spotting unit 3, and the subsequent operation is controlled.

  When the measurement item is colorimetric measurement, the dry analytical element 12 is transported in a state where the element presser 64 is positioned at the spotting portion, and then the sample tray 2 is rotated to rotate the spotting nozzle 45. The nozzle tip 14 of the tip rack 19 is moved downward and attached to the spotting nozzle 45. Subsequently, the specimen container 11 is moved, the spotting nozzle 45 is lowered, the specimen is sucked into the nozzle tip 14, the spotting nozzle 45 is moved to the spotting part 3, and the specimen is spotted on the dry analytical element 12. .

  Simultaneously with the spotting nozzle 45 spotting the specimen, an ultrasonic vibration is generated in the piezoelectric element 60 for several seconds by a piezo driver (not shown). Then, the generated ultrasonic vibration is propagated to the development layer 12b1 via the mount 12a (support member 12a) of the dry analytical element 12. The specimen is promoted to move in the spreading layer by the ultrasonic energy, and spreads uniformly in the whole spreading layer 12b1 of the dry analytical element 12, and then the uniformly spread specimen passes through the reflection layer 12b2 and reaches the reaction layer 12b3. It reaches uniformly, and the reagent in the reaction layer reacts with the corresponding component in the specimen.

  Then, the colorimetric dry analytical element 12 on which the specimen is spotted is inserted into the first incubator 4. Next, after the element chamber 91 is rotated and held at a constant temperature for a predetermined time, the inserted dry analysis element 12 is sequentially moved to the position of the photometric head 96, and the reflection optical density of the dry analysis element 12 is measured. After the measurement is completed, the measured dry analytical element 12 is pushed out to the center side and discarded. The measurement result is output, and the used nozzle tip 14 is removed from the spotting nozzle 45 by the tip discarding unit 9 and dropped downward and discarded, and the processing is completed. During the colorimetric measurement, in the second incubator 5, as described above, the lower block 71 is raised and the upper block 63 is preheated.

  Next, when the inspection item is a dilution request, for example, when the blood concentration is too high to perform an accurate inspection, the dry analytical element 12 is transported to the spotting position, and then the nozzle tip 14 is moved. The nozzle is attached to the spotting nozzle 45, and the spotting nozzle 45 is lowered to suck the sample into the nozzle tip 14. After the aspirated specimen is dispensed from the nozzle tip 14 into the mixing cup 16, the used nozzle tip 14 is removed. Next, a new nozzle tip 14 is attached to the spotting nozzle 45, and the diluent is sucked from the diluent container 15 into the nozzle tip 14. The sucked diluted liquid is discharged from the nozzle tip 14 to the mixing cup 16. Then, the nozzle tip 14 is inserted into the mixing cup 16 and agitation is performed by repeating suction and discharge. After agitation, the diluted specimen is sucked into the nozzle tip 14, the spotting nozzle 45 is moved to the spotting section 3, and the specimen is spotted on the dry analytical element 12. Thereafter, similarly, constant temperature holding, photometry, element discarding, result output and chip discarding are performed, and the process is terminated.

  Next, in the case of measuring the ion activity, the upper block 63 is moved to the spotting unit 3 as described above, and the electrolyte type dry analytical element 12 is transported to the spotting position. The nozzle tip 14 is attached to the nozzle 45 and the specimen is aspirated. Next, the nozzle tip 14 is attached to the other spotting nozzle 45 and the reference liquid is sucked from the reference liquid container 17. Next, the specimen is spotted on one liquid supply hole of the dry analytical element 12 by one spotting nozzle 45, and the reference liquid is spotted on the other liquid supply hole of the dry analytical element 12 by the other spotting nozzle 45. To wear. Then, the control unit drives the vibration unit in the same manner as described above to spot the sample on the dry analysis element 12 by the spotting nozzle 45 and simultaneously applies ultrasonic vibration to the dry analysis element 12 for several seconds.

  Then, the dry analytical element 12 on which the sample and the reference liquid are spotted is transferred to the second incubator 5 by the movement of the sliding frame 62 together with the upper block 63 from the spotting portion 3, and kept at a constant temperature by raising the lower block 71. Meanwhile, the ion activity is measured by the potential measuring probe 78. After the measurement is completed, the dry analytical element 12 after measurement is transferred to the discard hole 69 by the movement of the sliding frame 62 and discarded. Then, the measurement result is output, both the used nozzle tips 14 are removed from both spotting nozzles 45 and discarded, and the process is terminated.

  According to the present embodiment, in a biochemical analysis method for measuring a sample developed on a development layer on which a sample is developed, ultrasonic vibration is applied to the analysis chip supplied with the sample when the sample is supplied to the analysis chip. Therefore, when the specimen is developed in the development layer, vibration energy can be supplied to the specimen to promote the movement of the specimen, so that the specimen can be uniformly and rapidly developed on the development layer. In addition, since ultrasonic vibration is concentrated and applied to the analysis chip during a period of several seconds from the time when the specimen is supplied to the development layer, which is a period during which the specimen is developed in the development layer, the specimen is placed in the vibration direction. The movement can be promoted, the speed at which the specimen is spread on the spreading layer can be improved, and the specimen can be evenly spread on the spreading layer. Further, by setting the period for applying the ultrasonic vibration to several seconds, it is possible to efficiently promote the development of the specimen without extending the time required for the conventional biological analysis.

  In addition, when the present invention is applied to an analysis chip in which the sample is spotted on the analysis chip as in the above-described embodiment, particularly where uneven development is likely to occur, the sample supplied concentrated on a part of the development layer Since it is possible to rapidly develop the entire development layer, the effect of reducing the development unevenness is more remarkable.

  In addition, when a dry analytical element including a plurality of layers including a reagent layer corresponding to a measurement item for a specimen is used as an analysis chip as in the above embodiment, a uniformly developed specimen is supplied to the reaction layer. Therefore, the reaction unevenness of the specimen in the reaction layer is reduced, the erroneous measurement of the dry analytical element due to the reaction unevenness can be suppressed, and an accurate analysis result can be obtained.

  In addition, when applying ultrasonic vibration through the holding member that holds the spreading layer as in the above embodiment, the ultrasonic vibration is uniformly applied to the entire dry analytical element through the holding member. This is preferable because the development to the development layer can be promoted.

  FIG. 6 is a view showing a modification of the piezoelectric element 60 of the present embodiment. As shown in FIG. 6, in the above embodiment, when ultrasonic vibration is applied to the dry analytical element 12, the surface of the piezoelectric element 60 covers the spotting hole 12c and / or the photometric hole 12d of the dry analytical element 12. 12 m may be positioned in close contact with the upper surface 12 h or the lower surface 12 k of the dry analytical element 12. In the above case, the spotting unit 3 further includes a known drive mechanism (not shown) that raises the first element presser 63 and a known drive mechanism (not shown) that slides the piezoelectric element 60 in the horizontal direction. Immediately after spotting on the analysis chip, the first element presser 63 is raised to create a gap between the first element presser 63 and the dry analysis element 12, and the piezoelectric element 60 is inserted into the gap to dry the piezoelectric element 60. Ultrasonic vibration can be applied to the upper surface 12 h of the analysis element 12. Similarly, a well-known drive mechanism (not shown) is also provided below the dry analytical element 12 to slide the further piezoelectric element 60 in the horizontal direction, and immediately after the sample is spotted on the analysis chip, the dry analytical element 12 is provided. An ultrasonic vibration can be applied from the piezoelectric element 60 to the dry analysis element 12 by inserting the piezoelectric element 60 at a position in contact with the lower surface 12k of the electrode.

  In such a case, since the measuring element 12b touches only the sealed space 12m covering the spotting hole 12c and / or the photometric hole 12d of the dry analytical element 12, evaporation of the specimen into the atmosphere when applying ultrasonic vibration is performed. Is particularly effective in the case of an analysis for measuring a component easily vaporized in a specimen.

  As described above, when the space in contact with the development layer is sealed by the piezoelectric element 60, the space in contact with the development layer can be quickly set as the sealed space 12 m at the same time as the piezoelectric element 60 is disposed in the dry analysis element 12. Moreover, it is not necessary to provide an extra part for sealing the space where the spreading layer is in contact. When the surface of the piezoelectric element 60 is contaminated by the specimen, the piezoelectric element 60 may be exchanged for each inspection item together with the dry analysis element 12.

  In addition, a sealing member such as a protective film for covering the spotting hole 12c and / or the photometric hole 12d of the dry analytical element 12 is separately provided, and the space where the development layer is in contact is sealed by a sealing member that covers the space where the development layer is in contact. May be. In this case, the ultrasonic vibration can be applied after the sealing member is disposed so as to cover the spotting hole 12c and / or the photometric hole 12d as soon as the specimen is spotted on the dry analytical element 12.

  In the above embodiment, the analysis method for spotting a sample on a dry analytical element has been described. However, the present invention is not limited to this, and ultrasonic vibration is applied to an analysis chip having a spreading layer for spreading a sample. It can be applied to various possible analysis methods. As a second embodiment, an example will be described in which the present invention is applied to a detection apparatus that uses a microchannel sensor chip used for biomeasurement as an analysis chip.

  In bio-measurement, the presence / absence and amount of an antigen (or antibody) as a substance to be detected are measured by detecting a biomolecular reaction such as an antigen-antibody reaction. For example, one of two substances that specifically bind to each other (for example, an antigen, an antibody, various enzymes, a receptor, etc.) is immobilized on a substrate, and the other substance in an amount corresponding to the amount of the substance to be detected is By binding to a fixed layer fixed on a substrate and detecting this binding reaction, the presence / absence and amount of the substance to be detected in the sample can be measured. Specifically, in order to detect an antigen that is a substance to be detected contained in a sample, an antibody that specifically binds to the antigen is immobilized on the substrate, and the sample is supplied onto the substrate to supply the antigen to the antibody. Then, a labeled antibody with a label that specifically binds to the antigen is added and bound to the antigen to form a so-called sandwich of antibody-antigen-labeled antibody. A sandwich method that detects signals from, and a competitive method that detects the signal from the label attached to the competitive antigen bound to the immobilized antibody by binding the labeled competitive antigen to the immobilized antibody competitively with the antigen Immunoassays such as these have been made.

  FIG. 7 shows a schematic configuration of a measuring apparatus according to the second embodiment of the present invention, and FIG. 8 is a schematic perspective view showing an appearance of a sensor chip according to the second embodiment. The measurement apparatus of the present embodiment is an apparatus that detects a biological substance using the above-described micro-channel sensor chip (hereinafter simply referred to as a sensor chip) 110 as an analysis chip. The sensor chip 110 will be described with reference to FIGS.

  The sensor chip 110 is detachable from the measuring apparatus main body, and as shown in FIGS. 7 and 8, a flow path member 112 having a micro flow path 111 through which a sample liquid flows, and a micro flow path 111. The sensor part 114 which fixes one substance 113 of the two substances that specifically bind to each other to the wall surface, and the upper plate member 117 fixed on the flow path member 112 And. A membrane filter 118 is formed in the flow path member 112 on the upstream side of the sensor unit 114, that is, on the left side in the drawing. As will be described later, the membrane filter 118 functions as a spread layer.

  In the present embodiment, an example in which an assay by a sandwich method is performed in an antigen-antibody reaction will be described, and the substance 113 will be described as an antibody that specifically binds to the antigen A1 that is a substance to be detected. The antibody 113 may be directly fixed to the wall surface of the microchannel 111. However, as will be described later, when fluorescence is enhanced by electric field enhancement by surface plasmon, a metal thin film is formed on the wall surface. The antibody 113 is immobilized on.

  As shown in FIG. 8, the upper plate member 117 communicates the sample liquid inlet 116a and the sample liquid outlet 116b opened on the upper surface, and the sample liquid inlet 116a and the upstream end of the microchannel 111. It has an opening 15a, and an opening 15b that allows the sample solution outlet 116b and the downstream end of the microchannel 111 to communicate with each other. The upper plate member 117 and the flow path member 112 are joined by, for example, ultrasonic welding.

  The flow path member 112 and the upper plate member 117 are made of a transparent dielectric material such as polystyrene, and are respectively molded by injection molding. The depth of the microchannel 111 is, for example, about 50 to 100 μm.

  In the sensor chip 110 of this example, a labeled antibody (not shown) is attached to the inner surface of the microchannel 111 on the upstream side of the region where the antibody 113 is fixed. The labeled antibody is composed of an antibody that specifically binds to an epitope different from that of the aforementioned antibody 113 and a fluorescent label for the substance to be detected. Here, as the fluorescent label, fluorescent fine particles comprising a large number of fluorescent dye molecules f and a light transmitting material enclosing the fluorescent dye molecules f are used.

The size of the fluorescent fine particles is not particularly limited, but is preferably about several tens of nm to several hundreds of nm. In this example, one having a diameter of about 100 nm is used. Specific examples of the light transmitting material include polystyrene, SiO _{2 and the} like, as long as they can encapsulate the fluorescent dye molecule f and transmit the fluorescence from the fluorescent dye molecule f to be emitted to the outside. There is no particular limitation. The labeled antibody in this example is constituted by surface-modifying the fluorescent label 122 with a smaller antibody.

  In the sensor chip 110 of this example, the membrane filter 118 provided further upstream than the labeled antibody has a pH adjuster attached to the inside of the membrane filter 118. This is because a pH adjusting agent for increasing the reaction efficiency of the antibody 113 is supplied to the specimen passing through the membrane filter 118.

Next, returning to FIG. 7, the measuring apparatus will be described. In this measuring apparatus, a total reflection condition is applied to the prism 130 on which the sensor chip 110 is placed via, for example, refractive index matching oil, and the bottom surface of the microchannel 111 (interface between the sensor chip 110 and the sample liquid). A light source 131 made of a semiconductor laser or the like that makes the excitation light L ₀ incident at an incident angle of: a communication pipe 133 whose one end communicates with the sample liquid outlet 116 b of the sensor chip 110 via the nozzle 32, and this communication pipe 133. A sample suction pump 134 having a suction port connected to the other end thereof, an open valve 135 interposed in the communication pipe 133, and fluorescence Lf emitted from the vicinity of the sensor part 114 of the sensor chip 110 as described later. And a photodetector 136 for detection.

  Furthermore, this measuring apparatus is arranged above the membrane filter 118 via the wall surface of the microchannel 111 and serves as a piezoelectric element (piezo element in this example) 140 as ultrasonic wave generating means, and this piezoelectric element 140 is excited. A piezoelectric driver 141 that applies a driving voltage to the piezoelectric element 140; and a waveform generator 142 that generates a waveform signal that defines the waveform of the driving voltage and inputs the waveform signal to the piezoelectric driver 141. Note that the ultrasonic wave generation means is not limited to the piezoelectric element, and other piezoelectric ceramics can be applied.

  Next, detection of a substance to be detected by this measuring apparatus will be described. Here, as an example, a case where an antigen A1 that may be contained in blood (whole blood) S as a sample solution is detected will be described. First, the whole blood S is injected into the sample liquid inlet 116a shown in FIG. 1, and the sample suction pump 134 is driven at the same time, the open valve 135 is set to open the communication tube 133, and the whole blood S is detected in the sensor chip 110. Are introduced into the microchannel 111. At this time, the piezoelectric element 140 is driven by the piezo driver 141, and ultrasonic vibration is applied from the piezoelectric element 140 toward the membrane filter 118 provided so as to cross the micro flow path 111.

  The application of ultrasonic vibration is preferably performed during part or all of the period from the start of passage of the whole blood S to the membrane filter 118 until the end of passage. Including the time when it reaches the filter 118 is particularly preferable because the speed at which the whole blood develops on the membrane filter 118 is high.

  The whole blood S introduced into the microchannel 111 includes blood cells (red blood cells, white blood cells, and platelets) H1 as schematically shown in FIG. 7, and can also include an antigen A1. When the whole blood S reaches the portion of the microchannel 111 where the membrane filter 118 is provided, the whole blood S receives ultrasonic vibration energy and is promoted to move. For this reason, the speed of passing through the membrane filter 118 of the microchannel 111 is increased, and the whole blood S is uniformly spread on the membrane filter 118, and the pH adjuster contained in the membrane filter 118 is uniformly supplied to the whole blood S. The

  Thereafter, the plasma contained in the whole blood S is mixed with the labeled antibody that is adsorbed and fixed to the microchannel 111. As a result, the antigen A1 binds to the antibody of the labeled antibody, the antigen A1 bound to the antibody binds to the antibody 113 of the sensor unit 114, and a so-called sandwich is formed in which the antigen A1 is sandwiched between the antibody 113 and the antibody. .

The antigen A1 thus adsorbed on the sensor unit 114 is detected as follows. The excitation light L ₀ emitted from the light source 131 is incident on the bottom surface of the microchannel 111 (the interface between the sensor chip 110 and the sample liquid) at an incident angle that is a total reflection condition. When the excitation light L ₀ is totally reflected in this way, evanescent light oozes out from the inner wall surface of the microchannel 111 to which the antibody 113 is fixed into the sample liquid S. At this time, if a fluorescent label is present in the area where the evanescent light oozes, the fluorescent label is excited to generate fluorescence Lf. The fluorescence Lf generated in this way is detected by the photodetector 136. Detecting the presence of the fluorescent label as described above means that the presence of the antigen A1 bound to the antibody 113 is detected. Therefore, based on the fluorescence detection signal of the photodetector 136, the presence or absence of the antigen A1 and the amount thereof can be detected.

  In the above embodiment, the multi-function generator WF1974 manufactured by NF Circuit Design Block Co., Ltd. was used as the waveform generator 142, and the bipolar power supply HSA4101 manufactured by the same company was used as the piezo driver 141. The voltage waveform generated by the waveform generator 142 was amplified by the piezo driver 141 and applied to the piezoelectric element 140, and an ultrasonic wave was oscillated by thickness vibration. The applied voltage (pp value) at this time was 10.0V.

  In the second embodiment, since the pH adjuster contained in the membrane filter 118 can be uniformly supplied to the specimen by uniformly spreading the specimen on the membrane filter 118, the preferable reaction conditions with uniform pH are set. The reaction between the antibody 113 and the specimen can be performed in the region where the antibody 113 is originally fixed, and a highly accurate analysis result can be obtained. In addition, the present invention uses various substances and reagents necessary for analysis, such as labeled antibodies in the second embodiment, or a pretreatment agent such as a pH adjuster as in the second embodiment as a membrane. The filter 118 includes an analysis chip that supplies various substances, reagents, or pretreatment agents to a specimen that passes through the membrane filter, and a membrane filter and a reagent layer in order from the upstream of the flow path. The sample is applied to each layer or sensor unit provided in the flow path, such as an analysis chip of a type in which the sample that is uniformly developed in the cross section of the flow path is supplied to the reagent layer provided downstream of the flow path. The present invention can be suitably applied to various analysis chips having a development layer for allowing them to pass uniformly.

  In biomeasurement, it is desired to enable measurement in a shorter time. When the present invention is applied to a flow path type sensor chip as in the second embodiment, a development layer It is preferable that the measurement time can be made efficient by shortening the time for the specimen to flow through. In addition, in order to manage the measurement conditions uniformly because the specimen that reaches the sensor section is biased when the specimen passes through the membrane filter in a biased manner as in the case where the sensor section is provided in the lower part of the flow path. It is not preferable. However, according to the present embodiment, since the specimen can be uniformly developed on the development layer, the specimen can be supplied uniformly to the sensor unit and the like, and the measurement result of the specimen component can be obtained with high accuracy.

  In the apparatus shown in FIG. 7, the piezoelectric element 140 may be further provided only below the membrane filter 118 of the sensor chip 110, and the piezoelectric element 140 may be provided only below the membrane filter 118. In the case where the piezoelectric element 140 is provided both above and below the membrane filter 118, it is more preferable because vibration can be applied more uniformly and reliably.

  The present invention is not limited to the above embodiment, and can be applied to any bioanalytical method for measuring a developed specimen by spreading the specimen uniformly and quickly on the development layer. As an example, a specific substance or a target substance comprising an antibody is detected by utilizing a specific reaction between an antigen and an antibody as described in Japanese Patent Application Laid-Open No. 2009-257819 filed by the present applicant. It can be suitably applied to an immunochromatographic method. Further, as described in the background art of Japanese Patent Application Laid-Open No. 2009-180516, which is an application of the present applicant, a plurality of DNA fragments are electrophoresed on a gel support containing a fluorescent dye, and then a plurality of DNA fragments are electrophoresed. The DNA of the DNA fragment is denatured, and then at least a part of the denatured DNA fragment is transferred onto a transfer support such as nitrocellulose by Southern blotting, and the DNA or RNA complementary to the target DNA is fluorescent. A probe prepared by labeling with a dye is hybridized with a denatured DNA fragment, only a DNA fragment complementary to the probe DNA or probe RNA is selectively labeled, and the fluorescent dye is excited by excitation light, resulting in fluorescence Nitrocellulose etc. are also used in the method of detecting the target DNA distribution on the transfer support by generating an image by detecting Transfer support can be effectively applied to the present invention to the step of transferring (corresponding to an analyte in the present invention) non-transcript (corresponding to the spreading layer in the present invention).

  In addition, the present invention is not limited to the above-described embodiments, and various vibration modes, vibration periods, piezoelectric element types, development layer types, and the like are arbitrarily selected according to the type of analysis chip, the purpose of analysis, and analysis conditions. It is applicable to.

  Further, the present invention is not limited to the above embodiments, and the present invention may apply ultrasonic vibration to the dry analytical element from one direction, or may apply ultrasonic vibration to the dry analytical element from a plurality of directions. When ultrasonic vibration is applied to the dry analytical element from a plurality of directions, development in a plurality of directions can be promoted, and the specimen can be uniformly developed at a higher speed and in a wider range.

  Each of the above embodiments is merely an example, and all of the above description should not be used to limit the technical scope of the present invention. In addition, various modifications or combinations of the system configuration, hardware configuration, processing flow, module configuration, specific processing contents, and the like in each of the embodiments described above may be made without departing from the spirit of the present invention. Are included in the technical scope of the present invention.

  Examples and comparative examples according to the present invention will be described.

(Analysis chip)
A colorimetric type dry analytical element (CRE slide manufactured by FUJIFILM Corporation) was prepared. Moreover, when performing the below-mentioned evaluation test, the dry analysis element of the same kind and the same production lot number was used for the Example and the comparative example each time. Moreover, in order to make it easy to generate | occur | produce reaction nonuniformity, the dry analytical element which passed the expiration date in the following evaluation test was used.

(Evaluation test)
A control solution QP-H (manufactured by FUJIFILM Corporation, lot number 22032, CRE average concentration: 5.1 mg / dL) was spotted on the dry analytical element by a method. Thereafter, the colorimetric reaction of the dry analytical element was observed, and the reaction unevenness was visually evaluated in three stages. A comparative example was prepared by spotting the control solution QP-H on 10 dry analytical elements without applying ultrasonic vibration to the dry analytical element.

(Example)
For the 10 dry analytical elements with the same lot numbers as the comparative example, immediately after the control solution QP-H is spotted on the dry analytical element, the piezoelectric element is placed on the upper surface of the dry analytical element (on the spotted hole side). The evaluation test was performed by applying ultrasonic waves. The ultrasonic generation conditions were as follows.
Piezoelectric element: PZT-Pb (Zr / Ti) O3-based PZT soft material (C-82: resonance frequency 13 MHz) manufactured by Fuji Ceramics Co., Ltd.
Waveform generator: Multifunction generator WF1974 manufactured by NF Circuit Design Block Co., Ltd.
Vibration mode of piezoelectric element: Thickness longitudinal vibration mode Frequency: 13 MHz
Ultrasonic wave application time: about 15 seconds

(Evaluation results)
FIG. 9 shows the results of the colorimetric reaction of Examples and Comparative Examples. (I) of FIG. 9 is a diagram showing a dry analytical element when an ultrasonic wave is applied, and (II) of FIG. 9 is a diagram showing a dry analytical element according to a conventional method without applying an ultrasonic wave. As compared with the comparative example, it can be visually recognized that the reaction unevenness is suppressed in the example. Note that (I) and (II) in FIG. 9 use dry analytical elements whose expiration date has passed in order to remarkably confirm the occurrence of reaction unevenness.

Table 1 shows the results of visual evaluation of Examples and Comparative Examples 10 times.
In Table 1, the meaning of each symbol is as follows.
○: No reaction unevenness. Δ: Slight unevenness of reaction is observed. X: Reaction unevenness is observed.

  As shown in Table 1 above, in the comparative example, slight reaction unevenness that was visible was generated 4 times, and remarkable reaction unevenness occurred 6 times, but no reaction unevenness occurred in the examples. . Further, in this example, ultrasonic vibration was applied in the thickness direction of the dry analytical element, but the effect of suppressing the reaction unevenness can be sufficiently obtained also in the direction perpendicular to the thickness direction of the dry analytical element in which reaction unevenness is likely to occur. It was confirmed.

DESCRIPTION OF SYMBOLS 1 Biochemical analyzer 2 Sample tray 3 Spotting part 6 Spotting mechanism 12 Dry analytical element (analysis chip)
12a Mount part 12b Measuring element 12b1 Expanding layer 12b2 Reflecting layer 12b3 Reaction layer 12b4 Transparent support layer 12c Spotted hole 12d Photometric hole 12m Sealed space 60 Piezoelectric element 110 Sensor chip (analysis chip)
111 Microchannel 112 Channel member 118 Membrane filter 140 Piezoelectric element 141 Piezo driver 142 Waveform generator

Claims (9)

A biochemical analysis method for measuring a specimen using an analysis chip having a spreading layer for spreading the specimen,
Supplying the sample to the analysis chip;
A biochemical analysis method comprising applying ultrasonic vibration to the analysis chip supplied with the specimen.   2. The biochemical analysis method according to claim 1, wherein the sample is supplied by spotting the sample on the analysis chip.   The biochemical analysis method according to claim 1 or 2, wherein a dry analysis element comprising a plurality of layers including reagent layers corresponding to a plurality of measurement items for the specimen is used as the analysis chip.   For the supply of the sample, the measurement of the flow channel member of the analysis chip in which a micro flow channel for circulating the sample is provided in the flow channel member, and a measurement unit is disposed in a part of the micro flow channel. The biochemical analysis method according to claim 1, wherein the biochemical analysis method is performed by supplying a specimen to the development layer provided on the upstream side of the flow path from the section.   The biochemical analysis method according to claim 1, wherein the ultrasonic vibration is applied by applying ultrasonic vibration to the analysis chip from a plurality of directions.   The biochemical analysis method according to claim 1, wherein the application of the ultrasonic vibration is performed by sealing a space in contact with the development layer.   The biochemical analysis method according to claim 6, wherein the space in contact with the development layer is sealed with a piezoelectric element for applying ultrasonic vibration.   The biochemical analysis method according to claim 6, wherein the space in contact with the development layer is sealed by a sealing member that covers the space in contact with the development layer. A biochemical analyzer that measures an analyte using an analysis chip that has a development layer for deploying the analyte,
Means for supplying the sample to the analysis chip;
A biochemical analyzer comprising: means for applying ultrasonic vibration to the analysis chip supplied with the specimen.