[go: up one dir, main page]

WO2007018187A1 - Instrument de mesure, kit de mesure employant cet instrument, procédé de mesure, dispositif de mesure, et procédé permettant de reproduire un oscillateur piézoélectrique - Google Patents

Instrument de mesure, kit de mesure employant cet instrument, procédé de mesure, dispositif de mesure, et procédé permettant de reproduire un oscillateur piézoélectrique Download PDF

Info

Publication number
WO2007018187A1
WO2007018187A1 PCT/JP2006/315614 JP2006315614W WO2007018187A1 WO 2007018187 A1 WO2007018187 A1 WO 2007018187A1 JP 2006315614 W JP2006315614 W JP 2006315614W WO 2007018187 A1 WO2007018187 A1 WO 2007018187A1
Authority
WO
WIPO (PCT)
Prior art keywords
measurement object
measurement
piezoelectric vibrator
sample
measuring instrument
Prior art date
Application number
PCT/JP2006/315614
Other languages
English (en)
Japanese (ja)
Inventor
Haruki Tsunoda
Akihito Tomita
Takaaki Nozaki
Original Assignee
Kyowa Medex Co., Ltd.
Citizen Holdings Co., Ltd.
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Kyowa Medex Co., Ltd., Citizen Holdings Co., Ltd. filed Critical Kyowa Medex Co., Ltd.
Publication of WO2007018187A1 publication Critical patent/WO2007018187A1/fr

Links

Classifications

    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N5/00—Analysing materials by weighing, e.g. weighing small particles separated from a gas or liquid
    • G01N5/02—Analysing materials by weighing, e.g. weighing small particles separated from a gas or liquid by absorbing or adsorbing components of a material and determining change of weight of the adsorbent, e.g. determining moisture content

Definitions

  • Measuring instrument measuring kit using the same, measuring method, measuring device, and method of regenerating piezoelectric vibrator
  • the present invention relates to a measuring instrument for measuring a measurement object contained in a sample to be analyzed such as a living body, food, or soil, and in particular, has a piezoelectric vibrator at a detection site of a reaction vessel, BACKGROUND ART Related to a measuring instrument effective in an aspect in which a reaction with an object to be measured is performed on the surface portion of the piezoelectric vibrator, a measuring kit, a measuring method, a measuring apparatus using the same, and a method for regenerating a piezoelectric vibrator
  • a piezoelectric vibrator such as a quartz vibrator
  • SA M Self Assembled Monolayer
  • a method of reusing a crystal resonator sensor after supporting a target substance on the crystal resonator is known.
  • Non-Patent Document 1 a method of reusing a crystal resonator sensor by separating smoke particles (adsorbed molecules) adsorbed on the crystal resonator using a hot air source
  • Patent Document 1 a method of reusing the crystal resonator sensor by making the surface of the crystal substrate in the crystal resonator sensor flat to facilitate cleaning of this surface
  • Patent (Ref. 2) a method of reusing the crystal resonator sensor by making the surface of the crystal substrate in the crystal resonator sensor flat to facilitate cleaning of this surface.
  • the SAM method has a technical problem that, for example, the process of generating a sensor film made of a predetermined target substance on the electrode surface of a crystal resonator is very complicated and time-consuming.
  • the quartz vibrator sensor uses an expensive gold electrode, so that it is desired to reuse the gold electrode.
  • All of the conventional methods for reusing a quartz crystal sensor are: a sensor film generated on the surface of the crystal oscillator electrode, or thermal treatment by a hot air source. Since the separation process is complicated, for example, by giving a detachment, a simple method for reusing a crystal resonator sensor is desired.
  • POCT point-of-care testing
  • Non-Patent Document 1 “Biochemistry” American Chemical Society, 1998, 37th, 5666- 5672
  • Patent Document 1 Japanese Patent Laid-Open No. 2000-275157
  • Patent Document 2 JP 2000-283905 A
  • the present invention has been made to solve the above technical problem, and in a mode in which a piezoelectric vibrator is disposed at a detection site of a reaction vessel, a piezoelectric vibrator sensor film can be easily generated.
  • Another object of the present invention is to provide a measuring instrument, a measuring kit, a measuring method, and a measuring apparatus that can be reused by easily regenerating the sensor.
  • the present invention relates to the following (1) to (29).
  • a measurement instrument for measuring a measurement object in a sample using a reaction vessel having a sample addition site and a detection site equipped with a piezoelectric vibrator, and for applying a magnetic field to the piezoelectric vibrator A measuring instrument comprising: a magnetic force generating member, wherein the carrier is magnetically supported on the piezoelectric vibrator by the magnetic force of the magnetic force generating member.
  • the reaction container is bonded to the measurement object or the measurement object analog, or the binder or the measurement object analog.
  • a measuring instrument comprising a binder or a labeled body supply site capable of supplying a labeled body formed by binding to an insoluble endoplasmic reticulum.
  • the reaction container is a flow cell type in which the measurement object and the support body can move along the flow path, and the flow path of the flow cell type reaction container A measuring instrument, wherein a sample addition site and a detection site are provided.
  • the reaction container is a flow cell type in which the measurement object, the support, and the label can move along the flow path.
  • a measuring instrument comprising a sample addition site, a detection site, and a binder or label supply site in the flow path of the reaction vessel.
  • a measuring instrument according to any one of (1) to (8), wherein a plurality of types of carriers are used.
  • a measuring kit comprising the measuring instrument according to any one of (1) to (13) and a carrier.
  • a measuring kit comprising the measuring instrument according to any one of (1) to (13), a support, and a binder or a label.
  • a reaction vessel having a sample addition site and a detection site equipped with a piezoelectric vibrator, and a trapper that specifically binds to a measurement object or a measurement object analog and magnetic particles are combined.
  • the measurement object in the sample is measured using the support, and the measurement object in the sample and the support are reacted in a reaction container, and the composite including the support is measured.
  • the sample reaction step to be generated, the magnetic force acting on the magnetic particles of the carrier, the carrier carrying step of magnetically carrying the magnetic particles on the piezoelectric vibrator, and the amount of the composite carried on the piezoelectric vibrator is piezoelectric
  • a measurement method comprising: reacting a labeled body formed by binding with a cell body to generate a complex including a carrier, a measurement object, and a binder; and a carrier, a measurement object, and a complex including the label. .
  • a reaction vessel having a sample addition site and a detection site equipped with a piezoelectric vibrator, and a trapper that specifically binds to a measurement object or a measurement object analog and magnetic particles are combined.
  • the measurement object in the sample is measured using the carrier, and the measurement object in the sample and the analog of the measurement object are bound to the insoluble endoplasmic reticulum in the reaction container.
  • the sample reacts with the carrier to form a complex containing the carrier, and the carrier carries the magnetic particles on the piezoelectric vibrator by applying a magnetic force to the magnetic particles of the carrier.
  • a body supporting step a frequency measuring step for measuring the amount of the composite supported on the piezoelectric vibrator as a change amount of the frequency of the piezoelectric vibrator, and a frequency of the piezoelectric vibrator measured in the frequency measuring step.
  • a measurement method comprising a concentration determination step for determining the concentration of a measurement object in a sample from a change amount of the sample and a calibration curve prepared in advance.
  • a reaction vessel having a sample addition site and a detection site equipped with a piezoelectric vibrator, a carrier formed by binding a measurement object analogue and magnetic particles, and a measurement object or measurement object A method for measuring a measurement object in a sample using a binder that specifically binds to an analog or a label formed by binding the binder to an insoluble endoplasmic reticulum.
  • the object to be measured and the object to be measured and the magnetic object are bonded to each other.
  • the carrier and the binder that specifically binds to the object to be measured or the object to be measured or the binder and the insoluble vesicle are reacted.
  • Support body Sample reaction process for generating a composite containing a binder or a support body-a label body, a support body support that applies magnetic force to the magnetic particles of the support body and magnetically supports the magnetic particles on the piezoelectric vibrator Process, supported on piezoelectric vibrator Frequency measurement step of measuring the amount of the composite as a change amount of the vibration frequency of the piezoelectric vibrator, and the change amount of the vibration frequency of the piezoelectric vibrator measured in this frequency measurement step and a calibration curve prepared in advance And a concentration determination step for determining the concentration of the measurement object in the sample.
  • the reaction vessel is a flow cell type in which the measurement object and the support can move along the flow path, and the flow cell type reaction is performed.
  • a measuring apparatus comprising:
  • a method for reproducing a piezoelectric vibrator in measurement of a measurement object in a sample using a carrier and a piezoelectric vibrator comprising: a carrier and a measurement object carried on the piezoelectric vibrator by a magnetic force generating means.
  • a method for regenerating a piezoelectric vibrator comprising: measuring a composite body, and then dissociating the carrier by a magnetic field release means.
  • a support formed by binding a trapper that specifically binds to a measurement object or a measurement object analog and a magnetic particle, or a measurement object analog
  • a measuring instrument for measuring an object to be measured in a sample using a carrier bonded to magnetic particles, the reaction container having a sample addition site and a detection site equipped with a piezoelectric vibrator, and this A magnetic force generating member for selectively applying a magnetic field to the piezoelectric vibrator is provided, and the carrier is magnetically supported on the piezoelectric vibrator by the magnetic force of the magnetic force generating member.
  • a carrier functioning as a sensor film can be easily generated on the piezoelectric vibrator and can be removed. For this reason, in a measuring instrument equipped with a piezoelectric vibrator at the detection site, Thus, the piezoelectric vibrator sensor film can be easily generated, and the force can be reused by easily regenerating the sensor.
  • the measurement kit includes a measuring instrument and a carrier according to the present invention, and further includes a noinder or a label, the measurement object can be measured very easily.
  • a measuring instrument that can easily generate a piezoelectric vibrator sensor film and regenerate the sensor can be used to measure a large number of items in a sample. It is possible to measure quickly and accurately for each object.
  • the piezoelectric vibrator sensor can be easily reproduced.
  • FIG. 1 (a) is an explanatory plan view showing a basic configuration of a measuring instrument according to the present invention, and (b) is an explanatory view showing a detection site thereof.
  • FIG. 2] (a) to (c) are explanatory views showing the measurement principle by the measuring instrument of FIG.
  • FIG. 3 is an explanatory view showing a state after the reaction of the measuring instrument of FIG. 1 is completed.
  • FIG. 4 (a) and (b) are explanatory views showing another embodiment of the measuring instrument according to the present invention and the measurement principle thereof.
  • FIG. 5 is an explanatory diagram showing a measurement principle of measurement in another aspect by the measurement instrument of FIG. 1.
  • FIG. 6 (a) is an explanatory view showing an embodiment in which the present invention is applied to a measuring instrument including a flow cell type reaction vessel, and (b) is an arrow view seen from the M direction in (a).
  • FIG. 7 (a) is an explanatory view showing a more preferred embodiment in which the present invention is applied to a measuring instrument including a flow cell type reaction vessel, and (b) is an arrow view seen from the M direction in (a).
  • FIG. 8 (a) is an explanatory view showing a measuring method using the measuring instrument according to the present invention, and (b) is an explanatory view showing a measuring apparatus embodying the measuring method.
  • FIG. 9 is an explanatory view showing Embodiment 1 of a measuring apparatus to which the present invention is applied.
  • FIG. 10 is a schematic plan view of a measuring instrument used in Embodiment 1.
  • FIG. 11 is an explanatory diagram showing an equivalent circuit of the crystal resonator used in the first embodiment.
  • FIG. 12 (a) is an explanatory plan view showing a sensor substrate used in Embodiment 1
  • FIG. 12 (b) is an arrow view as viewed from the M direction in (a).
  • FIG. 13 (a) is an explanatory plan view showing the crystal resonator used in the first embodiment
  • FIG. 13 (b) is a cross-sectional explanatory view taken along line MM in (a).
  • FIG. 14 is an explanatory view showing Embodiment 2 of a measuring apparatus to which the present invention is applied.
  • FIG. 15 is a schematic plan view of a measuring instrument used in Embodiment 2.
  • FIG. 17 An explanatory diagram showing a third embodiment of a measuring apparatus to which the present invention is applied.
  • FIG. 19 is an explanatory view showing a modification of the crystal oscillation circuit used in the third embodiment.
  • FIG. 20 (a) is a schematic plan view showing a measuring instrument used in Embodiment 4 of the measuring apparatus to which the present invention is applied, and (b) is an arrow view seen from the M direction in (a). .
  • FIGS. 21 (a) and 21 (b) are explanatory diagrams showing an operation example of the measuring instrument according to the fourth embodiment.
  • FIG. 22 (a) is a schematic plan view showing a variation of the measuring instrument according to Embodiment 4, and (b) is (a
  • reaction vessel (flow cell type reaction vessel)
  • the representative embodiment of the measuring instrument according to the present invention specifically binds to the measurement object 10 (see FIG. 2 (a)) or the measurement object analog as shown in FIGS. L (a) and (b).
  • a measuring instrument for measuring the measurement object 10 in the sample using the carrier 11 formed by binding the trapper l ib and the magnetic particles 11a, the measurement object 10 in the sample (Fig. 2) (see (a)) includes a reaction container 2 that can be supplied, and the reaction container 2 includes a piezoelectric vibrator 6 provided at a detection site 5 where the measurement target 10 can be detected, and the piezoelectric vibrator 6.
  • a magnetic force generating member 7 that selectively and reversibly applies a magnetic field, and a carrier 11 that is magnetically supported on the surface of the piezoelectric vibrator 6 when the magnetic field is applied to the piezoelectric vibrator 6 are provided. It is characterized by.
  • FIGS. 4 (a) and 4 (b) another representative embodiment of the measuring instrument according to the present invention is, as shown in FIGS. 4 (a) and 4 (b), a carrier formed by combining a measurement object analogue 11c and magnetic particles 11a.
  • 11 is a measuring instrument for measuring the measurement object 10 in the sample, and includes a reaction vessel 2 to which the measurement object 10 in the sample can be supplied.
  • the piezoelectric vibrator 6 provided in the detection part 5 that can detect 10
  • the magnetic force generating member 7 that selectively and reversibly applies a magnetic field to the piezoelectric vibrator 6, and the magnetic field applied to the piezoelectric vibrator 6 And a carrier 11 that is magnetically supported on the surface of the piezoelectric vibrator 6.
  • the difference between the former and the latter is that the former carrier 11 is a magnetic particle 11a and a trapper l ib, whereas the latter carrier 11 is a magnetic particle 1 la and a measurement object analogue 1 lc. It is a point.
  • the trapper l ib or the measurement object analog 11c is used alone. These are not carried directly on the surface of the piezoelectric vibrator 6 but are carried on the surface of the piezoelectric vibrator 6 in the form of a carrier 11 combined with the magnetic particles 1 la.
  • the magnetic force generating member 7 also has a means for releasing the magnetic field applied to the piezoelectric vibrator 6.
  • a typical embodiment of the reaction vessel 2 includes one having a sample addition site 4 where a sample is usually added (see FIGS. 6 (a) and 6 (b)).
  • a labeled body 13 in which a binder 12 or a binder 13b or a target object analog 13c that specifically binds to a measurement object 10 is bound to an insoluble endoplasmic reticulum 13a can be supplied.
  • a binder Z labeled body supply site 8 see FIG. 2 (b) (c), FIG. 5, FIG. 7 (a) (b)).
  • the measuring instrument 1 of the present invention includes not only a flow cell type reaction vessel as long as it includes the reaction vessel 2, but also includes a well plate type reaction vessel.
  • the binder Z labeled body supply site 8 is the binder 12 or the labeled body 13 (see FIGS. 2 (b) and (c)). ) Can be selected as long as it can be supplied! /.
  • the binder 12 or the labeling body 13 is held in advance in the flow path 3 of the flow cell type reaction vessel 2, the binder 12 or the labeling body 13 is held in a part of the flow path 3, It can be moved with the added sample.
  • it may be provided downstream of the sample addition site 4 in the flow path 3 of the flow cell type reaction vessel 2.
  • the sample addition site 4 may also be used.
  • a detection site 5 in the flow path 3 of the flow cell type reaction vessel 2 is used. It is preferable to provide an absorption site 9 on the downstream side.
  • a typical example of the piezoelectric vibrator 6 is a quartz crystal vibrator.
  • the magnetic force generating member 7 includes a wide range of members that generate magnetic force, but a permanent magnet, an electromagnet or the like is typically used. [0025] ⁇ Element description>
  • the sample that can be used in the present embodiment is not particularly limited, and examples thereof include biological samples such as whole blood, plasma, serum, spinal fluid, saliva, amniotic fluid, urine, sweat, spleen, and tears.
  • biological samples such as whole blood, plasma, serum, spinal fluid, saliva, amniotic fluid, urine, sweat, spleen, and tears.
  • these samples or those derived from stool, food or soil can be used as samples by diluting, concentrating or extracting by adding an aqueous medium.
  • the aqueous medium is not particularly limited as long as it dissolves the above-described sample or label. Force buffers such as deionized water, distilled water, and buffer solutions are preferable.
  • the buffer used in the buffer is not particularly limited as long as it has a buffering capacity.
  • pH 1 to: L 1 For example, lactate buffer, citrate buffer, acetate buffer, succinate buffer, phthalate buffer , Phosphate buffer, triethanolamine buffer, diethanolamine buffer, lysine buffer, barbitur buffer, tris (hydroxymethyl) aminomethane buffer, imidazole buffer, malate buffer, oxalate buffer Agents, glycine buffer, borate buffer, carbonate buffer, glycine buffer, Good buffer, and the like.
  • Examples of good buffering agents include 2-morpholinoethanesulfonic acid (MES), bis (2-hydroxyethyl) iminotris (hydroxymethyl) methane (Bis-Tris), N- (2-acetamido) iminoniacetic acid (ADA), Piperazine-N, N, monobis (2-ethanesulfonic acid) (PIPES), N- (2-acetamido) 2-aminoethanesulfonic acid (ACES), 3 morpholino-2-hydroxypropanesulfonic acid (MOPSO), N , N Bis (2-hydroxyethyl) -2-aminoethanesulfonic acid (BES), 3-morpholinopropanesulfonic acid (MOPS), N- [Tris (hydroxymethyl) methyl] 2 aminoethanesulfonic acid (TES) ), 2- [4 (2 Hydroxyethyl) 1-piperadi-l] ethanesulfonic acid (HEPES), 3- [N, N bis (2 hydroxy
  • foods and soils that have been pretreated can be used as samples.
  • the pretreatment of food and soil includes, for example, extraction of ingredients in food and soil with an appropriate solvent, chemical modification, and the like.
  • the solvent include the above-mentioned aqueous media, organic solvents such as acetolyl, hexane, methanol, ethanol, dichloromethane, chloroform, and acetone.
  • Examples of the chemical modification include structural conversion of components in food and soil with chemical reagents.
  • the measurement object in the present embodiment is not particularly limited as long as it binds to a specific substance, for example, a component measured using an antigen-antibody reaction, a component measured using an enzyme reaction, and other specificities.
  • a component measured using an antigen antibody reaction for example, a component measured using an antigen-antibody reaction, a component measured using an enzyme reaction, and other specificities.
  • Ingredients measured by a chemical reaction can be mentioned, but a component measured using an antigen antibody reaction is preferred.
  • Components measured by antigen-antibody reaction include, for example, IgG, IgM, IgA, IgE, apoprotein AI, apoprotein ⁇ , apoprotein ⁇ , apoprotein ⁇ , rheumatoid factor, D-dimer, oxidized LDL, glycated LDL , Glycoalbumin, adiponectin, T3, ⁇ 4, drug (anti-tencan, etc.), C-reactive protein (CRP), cytodynamic ins, a-fetoprotein (AFP), cancer fetal antigen (CEA) ⁇ CA19 — 9, CA— 125, PIVKA- II (Protein induced by vitamin
  • biological components measured using an enzymatic reaction include glucose, 1,5-anhydroglucitol, hemoglobin Alc, glycoalbumin, fucose, urea, uric acid, ammonia, creatine, total cholesterol, free cholesterol, and high-density lipoprotein.
  • Cholesterol in protein (HDL—C), cholesterol in low density lipoprotein (LDL—C), cholesterol in very low density lipoprotein (VLDL—C), cholesterol in remnant-like lipoprotein (RLP—C), Triglyceride, phospholipid, total protein, albumin, globulin, pyrilvin, bile acid, sialic acid, lactic acid, pyruvic acid, free fatty acid, cell mouth plasmin, alanine aminotransferase (ALT), aspartate aminotransferase (A ST), Creatine phosphokinase (CPK) Over peptidase (PK), amylase, lipase, cholinesterase, I Monteagle Tamil trans peptidase, leucine aminopeptidase Daze, L-lactic acid dehydrogenase (LDH), aldolase, alkaline phosphatase, acid phosphatase, New ⁇ cetyl Darco Sami - Daze, Guanaze And monoamine
  • Examples of other components to be measured by specific binding include methods using nucleic acids, lectins and the like.
  • DNA or RNA encoding cancer genes such as ras, cancer suppressor genes such as p53, peptide nucleic acids, and abutama 1, glycoprotein and the like.
  • the analog of the measurement object is the measurement pair in the sample with respect to the binder or trapper.
  • the measurement object itself, a substance containing an epitope for a noinder or trapper, and the like.
  • a typical embodiment of the reaction vessel 2 is a flow cell type.
  • the flow cell type reaction vessel 2 widely includes those having a flow path 3 through which a sample flows, and is applied to immunochromatography, liquid chromatography, microchemical systems, and the like.
  • immunochromatography the sample solution added to the sample addition site moves through the membrane by capillary action and is measured at the detection site.
  • liquid chromatography a sample containing a measurement object is injected into a sample addition site installed in a flow cell through which liquid flows by a pump, and measurement is performed at a detection site.
  • a micro chemical system is an analytical sensor that integrates chemical devices such as semiconductor integrated circuits by processing minute flow paths on a substrate such as glass or plastic using micro processing technology.
  • Examples of the material of the flow cell type reaction vessel 2 include plastic, silica, ceramics, glass, metal, graphite, resin, and porous membrane.
  • reaction vessel a fixed cell type such as a well plate can be selected as appropriate.
  • the flow path 3 installed in the flow cell type reaction vessel 2 holds the measurement object, binder, and label together with the sample, flows without adsorption, and can form at least the sample addition site 4 and the detection site 5
  • plastic, silica, ceramics, glass, metal, graphite, porous membrane, etc. are preferable.
  • the material of the porous membrane include glass fiber, cellulose, nylon (registered trademark), crosslinked dextran, various chromatographic papers, nitrocellulose, and the like, and nitrocellulose is preferable.
  • the channel diameter is preferably lnm to 10 cm, more preferably lOOnm to lcm, and particularly preferably 1 ⁇ m to 2 mm.
  • At least one flow path 3 of the flow cell type reaction vessel 2 may be used. From the viewpoint of expanding the range, it is also possible to provide multiple lines of flow paths 3 and provide at least the sample addition site 4 and the detection site 5 in the flow path 3 of each line.
  • the sample addition site 4 is a site for adding the sample to the flow cell type reaction vessel 2.
  • Examples of the material of the sample addition site 4 include glass fiber, cellulose, nylon, cross-linked dextran, various chromatographic papers, and nitrocellulose. Nitrocellulose is preferable.
  • the binder Z label supply part 8 is a part for supplying a binder or a label, and the sample addition part 4 is also used as the binder Z label supply part 8, and the binder Z label is supplied from the same part as the sample addition part 4. It is also possible to supply a point force different from the sample addition site 4. In this case, the binder Z label can be added from the noinder Z label supply site 8, but the binder Z label may be held in the member.
  • Examples of the material of the Norder Z labeled substance supply site 8 include glass fiber, cellulose, nylon, cross-linked dextran, various chromatographic papers, nitrocellulose, and the like, which are the same as or different from those of the sample addition site 4.
  • the binder 12 in the present embodiment is present in a state where it can move in the flow path 3, and can be bound to magnetic particles.
  • antibodies and antibodies that specifically bind to the antigen and the antigen, saccharides and lectins for the saccharides, DNA and DNA complementary to the DNA, etc., each of which is used it can.
  • the labeled body 13 is composed of a measurement object analog or binder 13b and an insoluble vesicle 13a, and transmits information depending on the amount of the complex containing the carrier 11 formed on the piezoelectric vibrator 6.
  • a material in which the frequency change of the piezoelectric vibrator 6 is increased by the marker 13 is preferable.
  • the object recognition site on the label 13 The measurement object recognition site in the carrier 11 may be the same, but is preferably different.
  • insoluble vesicle 13a that binds to the measurement object analog or the binder 13b as long as the complex formed on the piezoelectric vibrator 6 can be detected as the amount of change in the frequency. What can increase the change in the frequency of the piezoelectric vibrator 6 having a large mass is preferable.
  • insoluble vesicles having a large mass include metal colloids and latex, and examples of metal colloids include gold colloids and silver colloids.
  • magnetic particles can also be used as the insoluble vesicle.
  • the particle size of the insoluble endoplasmic reticulum is preferably 0.1 to: LOOOO nm 1 to: 5 to 500 nm, more preferably LOOOnm.
  • the concentration of the insoluble endoplasmic reticulum used is preferably 0.001% to 10%, more preferably 0.01% to 5%, and particularly preferably 0.1% to 1%.
  • Insoluble endoplasmic reticulum can also be used in which the surface of the insoluble endoplasmic reticulum is coated with a hydrophilic protein such as bovine serum albumin or a polymer compound such as polyethylene glycol (PEG) or polyvinylpyrrolidone (PVA). .
  • the insoluble endoplasmic reticulum and the binder or the analog to be measured may be physically bonded or may be chemically bonded.
  • physical bonds include non-covalent bonds such as physical adsorption.
  • chemical bond include a covalent bond.
  • non-covalent bonds include electrostatic bonds, hydrogen bonds, hydrophobic bonds, and coordinate bonds.
  • the detection part 5 is preferably a quartz crystal vibrator as the piezoelectric vibrator 6 that is not particularly limited as long as it has the piezoelectric vibrator 6 that is a sensor.
  • the piezoelectric vibrator 6 is not particularly limited as long as it is made of a crystal having a piezoelectric effect.
  • crystals include crystals such as crystal, Rossiel salt, and electron stone, lithium tantalate (
  • Quartz is preferable.
  • the carrier 11 is composed of magnetic particles 1 la and a trapper 1 lb or a measurement object analogue 1 lc, and is not particularly limited as long as it is supported on the piezoelectric vibrator 6 by a magnetic force, but the trapper 1
  • the measurement object recognition site of the trapper l ib constituting the support 11 and the measurement target recognition site of the binder or label I prefer to be the same, but different! /.
  • the magnetic particles in the present embodiment are ferrite or magnetite (magnetite, Fe SO)
  • the particle size of the magnetic particles is preferably 1 to: LOOOOOnm force S, more preferably 10 to: LOOOOnm force S, and particularly preferably 100 to 500 Onm.
  • the concentration of magnetic particles supplied to the flow path as a labeling body is preferably 0.001% to 10%, more preferably 0.01% to 5%, and particularly preferably 0.1% to 1%. That's right.
  • magnetic particles whose surface is coated with a hydrophilic protein such as bovine serum albumin or a polymer compound such as polyethylene glycol (PEG) or polybulurpyrrolidone (PVA) can be used.
  • the trapper is not particularly limited as long as the trapper specifically binds to an object to be measured and is fixed on the crystal unit 25 together with magnetic particles.
  • Examples of the method for binding the trapper or the analog to be measured to the magnetic particle include physical bonding and chemical bonding.
  • physical bonds include non-covalent bonds such as physical adsorption.
  • Examples of the chemical bond include a covalent bond.
  • Examples of non-covalent bonds include electrostatic bonds, hydrogen bonds, hydrophobic bonds, and coordinate bonds.
  • Examples of the method of binding the trapper or the measurement object analogue to the magnetic particles by covalent bonding include a method of binding via a crosslinking agent using a crosslinking agent such as bivalent dartalaldehyde.
  • the magnetic force generating member 7 also has means for releasing the magnetic field applied to the piezoelectric vibrator 6.
  • the magnetic force generating member 7 is not particularly limited, such as a permanent magnet or an electromagnet. However, it is preferable that the magnetic force generating member 7 can generate a magnetic field reversibly so that the magnetic force can be applied or released as necessary. .
  • the magnetic field application range can be changed reversibly by making the position of the magnet relative to the surface of the piezoelectric vibrator 6 variable. It suffices to turn on / off the power supply to the coil.
  • the arrangement of the magnetic force generation member 7 is not particularly limited as long as the magnetic field can be selectively and reversibly formed on the piezoelectric vibrator 6.
  • the magnetic force generating member 7 may be present in the flow path 3, but if the viewpoint is such that the flow of the sample is not impaired, the flow cell type reaction vessel 2 It is preferably provided outside the channel 3.
  • the absorption site 9 is a site provided in the flow cell type reaction vessel 2 for absorbing the unreacted sample and label, and is preferably located downstream of the sample addition site 4 and the detection site 5.
  • the absorption site 9 can absorb unreacted components that have passed through the detection site 5 as long as it can absorb unreacted components (unreacted sample or label). The influence of the component on the detection site 5 can be reduced.
  • an absorptive polymer compound can be used as the absorption site 9.
  • the polymer compound include cellulose, glass fiber, cotton, polyurethane and the like.
  • the absorption part 9 can use a forced discharge means such as a pump.
  • washing liquid The cleaning liquid is not particularly limited as long as it can wash the sample components and the label that cannot react on the piezoelectric vibrator 6, but an aqueous medium is preferable. In particular, the above-mentioned aqueous medium containing a surfactant is more preferable.
  • the buffer used for the buffer is not particularly limited as long as it has a buffer capacity.
  • the surfactant is not particularly limited as long as it has a surfactant effect, and examples thereof include a cationic surfactant, an anionic surfactant, an amphoteric surfactant, and a nonionic surfactant. Nonionic surfactants are preferred.
  • Nonionic surfactants include polyoxyethylene sorbitan monolaurate (Tween 20) and polyoxyethylene octyl ether (TritonX-lOO).
  • concentration of the surfactant is not particularly limited, but is preferably 0.001 to 20%, more preferably 0.01 to 10%, and particularly preferably 0.05 to 1%.
  • the method for controlling the temperature of the reaction liquid in the flow cell type reaction vessel 2 is not particularly limited as long as the temperature can be controlled by heating the entire bottom surface force of the flow cell type reaction vessel 2, but for example, the temperature using a Peltier element Control.
  • the measurement object recognition site in the binder 12 or the label 13 may be the same as the measurement object recognition site in the carrier 11 (trapper l ib), but differently. I prefer to do that!
  • the reaction at the detection site 5 it is necessary to remove the reaction product generated on the piezoelectric vibrator 6 after the reaction is completed.
  • the magnetic interaction magnetic force or magnetic attractive force
  • the support 11 disengages from the surface force of the piezoelectric vibrator 6, the reactant on the piezoelectric vibrator 6 is released, and the surface of the piezoelectric vibrator 6 can be easily reset.
  • the support 11 is composed of the magnetic particles 1 la and the measurement object analogue 1 lc
  • the support 11 is supported on the piezoelectric vibrator 6.
  • the supported body 11 reacts competitively with the measurement object 10 with respect to the binder 12 or the label body 13.
  • a measurement object-like substance binder or a complex having a label strength is generated.
  • the measurement object analogue 11c reacts competitively with the measurement object 10, so if the reaction is accelerated with the measurement object analogue 11c, the corresponding amount in the sample is increased.
  • the number of measurement objects 10 is small, and the opposite reaction tendency means that there are many measurement objects 10 in the sample. Therefore, it is possible to indirectly increase the measurement sensitivity of the measurement object 10 by accurately measuring the amount of reaction with the measurement object analog 11c.
  • the magnetic field action by the magnetic force generating member 7 is solved in the same manner as in FIG. 3 in order to remove the reaction product generated on the piezoelectric vibrator 6 after the reaction is completed. It can be removed.
  • a carrier 11 for example, magnetic particles 1 la + trapper ib
  • a label 13 for example, measurement object analogue 13c
  • the measurement object 10 in a competitive manner, a composite body having a carrier measurement object or a measurement object analog force is generated on the piezoelectric vibrator 6.
  • the measuring instrument 1 in which the carrier 11 is a magnetic particle 11a and a trapper l ib.
  • the sample addition site 4 and the piezoelectric vibration A reaction vessel 2 having a detection site 5 with a child 6 and a carrier 11 formed by binding a trapper l ib that specifically binds to a measurement object or a measurement object analog and a magnetic particle 11a.
  • a sample reaction step for generating a magnetic material, a magnetic material 11a of the carrier 11 is made to act on a magnetic force, and a magnetic material 11a is magnetically supported on the piezoelectric vibrator 6 and a composite supported on the piezoelectric vibrator 6
  • the frequency measurement process for measuring the amount of the body as the amount of change in the frequency of the piezoelectric vibrator 6, and the piezoelectric measured in this frequency measurement process
  • a previously prepared calibration curve and the frequency of variation of Doko 6 as long as it includes a concentration determination step of determining the concentration of the measuring object 10 in the sample.
  • a washing step for the detection site 5 is included between the sample reaction process and the frequency measurement step, and unnecessary components at the detection site 5 are removed. It is preferable.
  • a sample addition site 4 and a piezoelectric vibrator 6 are provided.
  • the reaction vessel 2 having the detection site 5 and the support 11 formed by binding the trapper 11b specifically binding to the measurement object or the measurement object analog and the magnetic particle 11a are used.
  • the sample reaction step of reacting with the body 11 to generate a complex containing the label body, the magnetic particles 11a of the carrier 11 acting magnetically, and the magnetic particles 11a are magnetically supported on the piezoelectric vibrator 6
  • Supporting process Measures the amount of composite supported on the piezoelectric vibrator 6 as the amount of change in the frequency of the piezoelectric vibrator 6
  • a frequency measurement step and a concentration determination step for determining the concentration of the measurement object 10 in the sample from the amount of change in the frequency of the piezoelectric vibrator 6 measured in the frequency measurement step and a calibration curve prepared in advance. If you do,
  • FIG. 4 and FIG. 8 (a As shown in (b) , A reaction vessel 2 having a detection part 5 provided with a sample addition part 4 and a piezoelectric vibrator 6, a carrier 11 formed by combining a measurement object analogue 11c and a magnetic particle 11a, and a measurement object 10 or A method for measuring a measurement object 10 in a sample using a binder 12 that specifically binds to a measurement object analog or a labeled body 13 formed by binding the binder 13b and an insoluble vesicle 13a, In reaction container 2, specifically binds to the measurement object 10 or the measurement object analogue in the reaction object 10 and the measurement object analog 1 lc and magnetic particles 1 la.
  • Magnetic particle 11a magnetically piezoelectric vibrator A carrier supporting process for supporting the piezoelectric vibrator 6, a frequency measuring process for measuring the amount of the composite supported on the piezoelectric vibrator 6 as a change in the frequency of the piezoelectric vibrator 6, and this frequency measuring process
  • a concentration determining step for determining the concentration of the measurement object 10 in the sample from the amount of change in the frequency of the piezoelectric vibrator 6 measured in step 1 and a calibration curve prepared in advance may be provided.
  • the supply step includes the step of supplying the sample and the binder 12 or the label 13 from the sample addition part 4. It may be added at the same time or before and after.
  • the supply step Any sample may be used as long as the sample is added from the sample addition site 4 and the binder 12 or the label 13 held in advance in the Noinder Z label supply site 8 moves together with the sample.
  • any plurality of detection part 5 may be provided corresponding to the above, and the detection site 5 may be set at an arbitrary location. According to this aspect, it is possible to cause the magnetic fields generated by the plurality of magnetic force generating members 7 to act separately, and to allow only the magnetic field acting region to function as the detection portion 5.
  • the above-described measuring instrument 1 that supplies the analyte 13c to the detection site 5 with the labeled body 13 bound to the insoluble endoplasmic reticulum 13a and the piezoelectric vibrator 6 at the detection site 5 of the measurement instrument 1 were generated.
  • Trapper--measurement object or trapper--measurement object--binder or trapper--one measurement object Composite consisting of a label or measurement object analogue Non-one or measurement object analogue
  • the frequency measuring means 15 that measures the amount of the complex as the labeled body force as the amount of change in the vibration frequency of the piezoelectric vibrator 6, and the amount of change in the vibration frequency of the piezoelectric vibrator 6 that is measured by the vibration frequency measuring means 15 and created in advance. Measurement in the sample from the measured calibration curve That a density determining means 16 for determining the concentration of an object 10 and the like.
  • the measuring instrument 1, the frequency measuring means 15, and the concentration determining means 16 may be provided separately, or the measuring instrument 1 is provided with the frequency measuring means 15 and the concentration determining means 16. You may make it provide integrally by incorporating.
  • FIG. 9 is an explanatory view showing Embodiment 1 of the measuring apparatus to which the present invention is applied.
  • the measurement apparatus includes a measurement instrument 20 for measuring a measurement object in a sample, and an analysis processing apparatus 100 that performs analysis processing based on a sensor output from the measurement instrument 20.
  • the measuring instrument 20 has a flow cell type reaction vessel 21 in which a measurement object in a sample can move along a flow path 22 as shown in FIG. 9 and FIG.
  • the flow cell type reaction vessel 21 is sealed with a cover 30 so that a sensor substrate 24 of a QCM (Quartz Crystal Microbalance) sensor 23 is the bottom of the vessel and a flow path 22 with a predetermined gap is secured on the sensor substrate 24.
  • a liquid inflow opening 31 is formed in a part of the cover 30 of the flow cell type reaction vessel 21 located upstream of the flow path 22 and a liquid outflow is provided in a part located downstream of the flow path 22. Opening 32 is established.
  • a sample container 34 is connected to one liquid inflow opening 31 via a tube 33 and functions as a sample addition site A, and the other liquid outflow opening 32 via a tube 35.
  • the syringe pump 36 is connected to function as an absorption site D.
  • the label added from the sample addition site A is supplied at the same time as the sample or after the sample is added.
  • the QCM sensor 23 has a quartz resonator 25 mounted on the surface of the sensor substrate 24, and a portion of the flow path 22 corresponding to the surface of the quartz resonator 25 is detected. It is designed to function as C.
  • a magnet 40 made of, for example, a permanent magnet is installed on the back surface of the sensor substrate 24 at a position where the crystal unit 25 is mounted!
  • the magnet 40 is detachable, and when the magnet 40 is installed, the position is adjusted so that the surface portion of the crystal unit 25 is disposed in the magnetic field application region by the magnet 40.
  • a carrier 11 (magnetic particle l la + trapper l ib or measurement object analogue 11c) is carried on the surface of the crystal unit 25 by the magnetic field of the magnet 40 (FIG. 1 or FIG. 4). reference).
  • the sensor substrate 24 is placed on a constant temperature block 51 made of, for example, aluminum, which is maintained at a constant temperature by, for example, a Peltier element 52! Consideration is given to prevent changes in the resonance frequency.
  • the analysis processing device 100 calculates the concentration of the measurement object in the sample based on the frequency measurement circuit that measures the resonance characteristics of the crystal unit 25 and the information on the frequency measurement circuit force. It should be equipped with a concentration calculation circuit.
  • the resonance characteristics of the crystal unit 25 are measured using the network analyzer 101, and the measured resonance characteristic data is stored in a personal computer (PC).
  • Crystal vibration by taking in 102 and calculating An example is a configuration in which the resonance frequency of the element 25 is obtained and the calculation is performed on the resonance frequency force concentration using a calibration curve.
  • the resonance frequency from the resonance characteristics of the crystal unit 25 measure the resonance characteristics (frequency-admittance characteristics) near the resonance frequency of the crystal unit 25 using a network analyzer 101, etc. Find the equivalent circuit constants using mathematical means such as the least-squares method to match the admittance characteristics shown by.
  • an equivalent circuit of the crystal unit 25 can be expressed by a parallel connection of a series resonant circuit of an inductance Lx, a capacitance Cx, and a loss resistance Rx and a parallel capacitance Cp such as an electrode capacitance.
  • the admittance Y of the crystal unit can be obtained by Equation 1 and Equation 2.
  • J is a complex number (1 1) 1/2
  • ⁇ is an angular frequency
  • fo is a resonance frequency
  • ⁇ is a circularity
  • the resonance frequency fo of the quartz crystal is calculated by the equivalent circuit constant Lx , Formula from Cx
  • the personal computer 102 preferably has a display device 103 for displaying the concentration calculation result!
  • the crystal resonator 25 is, for example, an AT-cut crystal resonator having a thickness-shear vibration mode, and is preferably one that oscillates at a fundamental resonance frequency of 5 to 50 MHz. Further, one crystal unit 25 may be used as in the present embodiment, but a plurality of crystal units 25 may be used.
  • the structure of the sensor substrate 24 may be selected as appropriate, but an example is shown in FIGS. 12 (a) and 12 (b).
  • the sensor substrate 24 is a double-sided printed circuit board made of glass epoxy, for example.
  • plate 61 on the top surface, make two land patterns 62 to connect with electrodes 65 and 66 of crystal unit 25 and the vibrating part of crystal unit 25 not to contact printed circuit board 61.
  • a U-shaped pattern 63 for a pedestal is arranged.
  • the crystal unit 25 is arranged such that the electrode connection part of the crystal unit 25 is placed on the electrode connection land pattern 62, and is electrically connected with, for example, conductive silicone resin.
  • the periphery of the crystal unit 25 is, for example, a silicone adhesive 64. It is sealed with.
  • the wiring 67 from the electrode connecting land pattern 62 is drawn to the lower surface of the printed circuit board 61 through the through hole and extends to the connection terminal 68 to avoid short circuit due to the sample solution.
  • the front electrode 65 and the back electrode 66 are both provided with connection portions 69a so that they can be connected to the electrode connection land pattern 62 of the printed circuit board 61 on the back surface. It is electrically connected by the extraction electrode 69.
  • the front electrode 65 is routed by the extraction electrode 69 to the connection portion 69a on the back surface via the left side surface of the crystal unit 25.
  • the sensor substrate 24 constitutes a part of the flow cell type reaction vessel 21, but the present invention is not limited to this.
  • an adhesive is used. You should do it in a scientific way using chemicals!
  • the adhesive used is silicone conductive adhesive, epoxy conductive adhesive, anisotropic conductive film (ACF), anisotropic conductive paste (ACP), etc. for electrical connection, and waterproof and fixing. Silicone adhesives, epoxy adhesives, etc. are used.
  • the carrier 11 carried on the crystal unit 25 is detached from the external force of the reaction vessel 21 and the magnetic field generated by the magnet 40 is released. There is no particular restriction as long as you can leave 11.
  • a solution containing a carrier (magnetic particle + trapper or similar object to be measured) is added from the sample addition site A of the measuring instrument 20, and a magnet is placed on the surface of the quartz crystal resonator 25.
  • the magnetic field generated by 40 is applied, the carrier is supported on the crystal unit 25, and cleaning with the cleaning liquid is performed.
  • the sample and the binder (or labeled body) are added to the sample addition site A of the measuring instrument 20, and the measurement object and the binder (or labeled body) in the sample are the flow cell type of the measuring instrument 20.
  • the amount of the complex produced by specifically binding to the carrier is detected as the amount of change in the vibration frequency of the crystal unit 25, and the concentration of the measurement object in the sample is measured via the analysis processing device 100.
  • the conditions under which the measurement object in the sample reacts with the carrier (for example, a trapper) on the crystal unit 25 or the measurement object in the sample and the label in the reaction solution are not particularly limited as long as it allows specific reaction, but the reaction temperature is usually 0 ° C-100 ° C, preferably 10-60 ° C, more preferably 20-40 ° C. Is done.
  • the measurement time is usually 10 seconds to 10 hours, preferably 30 seconds to 5 hours, more preferably 1 minute to 1 hour.
  • the measurement using a standard substance with a known concentration for preparing a calibration curve may be repeated individually using the same measuring device 20, but using an apparatus configured by combining a plurality of measuring devices 20.
  • the measurement object 10 is measured directly or indirectly.
  • the magnetic field action by the magnet 40 is canceled and the cleaning liquid is allowed to flow, so that the carrier 11 is separated from the surface of the quartz oscillator 25 as shown in FIG. 3, for example.
  • the reactant on the crystal unit 25 is surely removed.
  • the measuring instrument 20 can be reused.
  • FIG. 14 shows Embodiment 2 of the measuring apparatus to which the present invention is applied.
  • the basic configuration of the measuring device is provided with a measuring instrument 20 and an analysis processing device 100 in substantially the same manner as in the first embodiment, but unlike in the first embodiment, it is applied to POCT (point-of-care testing). Considering the correspondence, it is designed to be a small and simple configuration. Components similar to those in the first embodiment are denoted by the same reference numerals as those in the first embodiment, and detailed description thereof is omitted here.
  • the entire apparatus is obtained by incorporating the analysis processing device 100 into the measuring instrument 20, and has two element forces, that is, a measurement unit S1 and a measurement result analysis processing unit S2.
  • the measurement unit S1 includes a QCM sensor 23 (a sensor substrate 24 on which a crystal unit 25 is mounted), a plastic upper cover 71, a lower cover 72, and various membranes 81 to 83.
  • the upper cover 71 is formed by hollowing out the channel 22 part, and is placed on the sensor substrate 24 in a detachable and watertight manner, and a gap formed between the two is defined as the channel 22 It is getting used.
  • a portion of the flow channel 22 is secured long, and on one side of the flow channel 22, there is a sample holding membrane 81 that functions as the sample addition site A.
  • a label (or binder) holding membrane 82 that functions as a label holding site (or binder holding site) B is sandwiched between the sample holding membrane 81.
  • an opening 75 for adding a sample solution with a pipette or the like is provided in the vicinity of the sample addition site A of the upper cover 71.
  • the opening 75 also functions as an auxiliary sample container.
  • an absorption membrane 83 is sandwiched on the downstream side of the detection site C, so that it functions as the absorption site D.
  • the label (binder) holding site B is a site for holding the label (binder) in the flow path 22 of the flow cell type reaction vessel 21 so that the label (binder) can move.
  • the material of the label (binder) holding membrane 82 at the label (binder) holding site B may be the same as or different from the material of the sample holding membrane 81 at the sample addition site A. Examples thereof include glass fiber, cellulose, nylon, cross-linked dextran, various chromatographic papers, nitrocellulose and the like, and nitrocellulose is preferable.
  • a water-absorbing polymer compound can be used as the absorption membrane 83 of the absorption site D.
  • the polymer compound include cellulose, glass fiber, cotton, polyurethane and the like.
  • a magnet 40 (for example, a rectangular ferrite magnet) is installed at the position where the crystal resonator 25 is mounted on the lower surface of the sensor substrate 24. The surface will be placed.
  • the mounting method of the sensor substrate 24 and the crystal resonator 25, the electrode drawing method, and the like are substantially the same as those in the first embodiment.
  • the sensor substrate 24 is assembled by bonding or fitting with the upper cover 71 and the lower cover 72 sandwiched therebetween. Further, since the crystal unit 25 is disposed in a space sealed by the upper and lower covers 71 and 72, it is not easily affected by environmental temperature changes or wind.
  • the measurement unit S1 and the measurement result analysis processing unit S2 can be mechanically connected and disconnected by a connector 85.
  • the wiring from the crystal unit 25 is also connected to the measurement result analysis processing unit S 2 via the connector 85.
  • the measurement result analysis processing unit S2 includes a frequency measurement circuit and a concentration calculation circuit, as in the first embodiment, but the crystal resonator is used as the frequency measurement circuit.
  • a configuration is possible in which 25 is connected to a crystal oscillation circuit 111 to oscillate in the vicinity of the resonance frequency, and the oscillation frequency is measured by the frequency counter 112.
  • a control device 113 such as a personal computer, a microprocessor, or a logic operation circuit can be used as the concentration operation circuit.
  • the measurement of the resonance frequency of the crystal unit 25 is performed at a frequency slightly higher than the resonance frequency when the crystal unit 25 is oscillated using the crystal oscillation circuit 111. Since it oscillates, it can be regarded as a resonance frequency. And the resonance frequency is obtained by measuring the oscillation output by the frequency counter 112, and the control device 113 using a CPU or the like uses the calibration curve data to determine the concentration of the measurement object from the resonance frequency. And the result can be displayed on the display device 114.
  • FIG. 16 shows an example of the crystal oscillation circuit 111, which is a Colpitts type crystal oscillation circuit using a CMOS inverter.
  • the wiring from the front electrode and the back electrode of the crystal unit 25 is connected to the crystal oscillation circuit 111 through the wiring of the sensor substrate 24.
  • the crystal unit 25 is connected to the ground via the input capacitor 221 and the output capacitor 222 to form a ⁇ -type feedback circuit, and this feedback resistor 223 is inserted between the input and output of the amplifier circuit composed of the CMOS inverter 224.
  • the signal is fed back and oscillated.
  • the C MOS inverter 224 is used as an inverting amplifier by connecting the input and output with a feedback resistor 223 of several ⁇ power M ⁇ .
  • the measuring device does not require large devices such as a syringe pump 36 or a network analyzer, and is a small-sized device suitable for POCT or the like.
  • a measuring device can be realized.
  • the measuring apparatus is a model using a single crystal unit as the QCM sensor 23. There is no particular problem with these models as long as the frequency drift of the crystal unit itself is negligible compared to the amount of frequency change due to complex coupling, where the frequency drift of the crystal unit itself is relatively small.
  • the frequency drift means a frequency fluctuation other than a frequency change based on specific binding of the target substance. For example, when the added sample solution comes into contact with the crystal resonator, the resonator itself or the ambient temperature is changed. Change, and the resonance frequency will change, and components other than the target substance contained in the sample solution will bind to the sensor film and cause a frequency change. It is.
  • the drift component is added to the measurement value, which may cause an error in the measurement value.
  • the crystal resonator that detects the target substance is called the test crystal resonator.
  • This reference crystal unit is attached to the test crystal unit at the detection site in the flow path, and the change in the resonance frequency of each test crystal unit is measured. By subtracting, it is possible to remove the influence of the drift component and reduce the measurement error.
  • FIG. 17 is a diagram showing the configuration of the third embodiment using a reference crystal resonator.
  • the measurement device according to the present embodiment is basically a reference crystal oscillator added to the second embodiment, and further measures the resonance frequency of the added reference crystal resonator and calculates the difference. This is an improvement of the measurement circuit 120 of the result analysis processing unit S2 (analysis processing device 100).
  • a reference crystal resonator 252 is installed downstream of the flow of the sample solution with respect to the test crystal resonator 251 provided at the detection site C in the flow path 22.
  • the method of mounting the reference crystal unit 252 can be the same as that of the test crystal unit 251!
  • the wiring from the reference crystal resonator 252 is connected to the measurement circuit 120 of the measurement instrument analysis processing unit S2 through the connector 85 through the wiring pattern of the sensor substrate 24.
  • the magnet 40 for applying a magnetic field is provided separately for the reference crystal resonator 252 and the test crystal resonator 251, but each magnet resonator 251, The magnet shape and installation position may be adjusted so that the magnetic field concentrates on the 252 electrode surfaces.
  • the positional relationship between the reference crystal unit and the test crystal unit can be either upstream or downstream in the case of series connection with the sample solution flow.
  • test crystal unit 251 and the reference crystal unit 252 are placed as close as possible so that they are equally affected by drift, the drift cancellation Preferred in terms of becoming more complete.
  • FIG. 18 is a diagram showing a configuration of the measurement circuit 120.
  • test crystal oscillator 251 and the reference crystal oscillator 252 are connected to the individual crystal oscillator circuits 121 and 122 and frequency counters 123 and 124, respectively. It is configured to continuously measure.
  • the Colpitts crystal oscillation circuit used in the second embodiment can be used as the crystal oscillation circuits 121 and 122.
  • the measured resonance frequencies are calculated by calculating the difference in the control device 125 so that the drift component is subtracted.
  • Reference numeral 126 denotes a display device that displays a calculation result by the control device 125.
  • the difference is calculated after the oscillation frequencies of the two are obtained by the frequency counters 123 and 124, but the difference frequency is created directly from the outputs of the two crystal oscillation circuits 121 and 122 using a logic circuit.
  • a method of measuring the difference frequency with a counter may be used.
  • FIG. 19 is a diagram showing another configuration of the measurement circuit 120.
  • the reference crystal resonator 252 and the test crystal resonator 251 are switched by the switching circuit 130 and are alternately oscillated by one crystal oscillation circuit 131 to measure the frequency.
  • the switching circuit 130 can be an electromagnetic relay or an electronic relay such as an analog switch.
  • Switching is performed by a switching signal generated in the control device 133.
  • the switching timing can be in the range of several 0.1s of force and several tens of seconds. If the switching time is short, the measurement accuracy of the frequency counter 132 will deteriorate. Conversely, if the switching time is long, the resonance frequency on the non-oscillating side Since the state of change disappears, a force of about 1 second per second is usually appropriate.
  • Reference numeral 134 denotes a display device that displays a calculation result by the control device 133.
  • force using separate crystal resonators for the test crystal resonator 251 and the reference crystal resonator 252 for example, two channels in which two sets of electrodes are provided on one crystal substrate.
  • a quartz crystal can also be used.
  • a 2-channel crystal unit is used, and the drift removal capability is superior compared to using two individual crystal units. It is suitable when higher accuracy and detection limit are required.
  • an influenza A virus can be detected with a first reference type 2-channel crystal resonator
  • an influenza B virus can be detected with a second reference type 2-channel crystal resonator.
  • 2-channel crystal unit In addition to the 2-channel crystal unit, it can also be applied to crystal units with 3 or more channels.
  • FIG. 20 shows a measuring instrument according to the fourth embodiment.
  • Quartz crystal unit 25 control crystal unit 25 is placed in channel 22 (22) of line n
  • control crystal unit 25 is a crystal unit on which a sensor film that specifically detects magnetic particles not forming a complex is formed. Can be used as an index to indicate whether the measurement itself has been completed normally, that is, whether measurement is possible.
  • n crystal resonators at the detection site are used as test crystal resonators, so that the same measurement object in n samples can be measured simultaneously. And n measurement objects in the same specimen can be simultaneously measured.
  • one of the multiple lines is used as the reference crystal unit 25.
  • crystals are formed at a plurality of locations (P to P) of the reaction vessel 21.
  • the vibrator 25 (25 to 25) and a plurality of carriers (A to 8) are supported by the magnet 40 (40 to 40).
  • the respective supporting bodies (A to A) are detached separately from the surface of the crystal unit 25. be able to.
  • the supports (A to A) may be different from each other, but two or more types may be the same.
  • the binder or the label may be different from each other, and two or more kinds of force may be the same.
  • sample addition site A is shared, but this is not limited to this. As shown in FIG. 22, the sample addition site for each channel is shown. A may be provided.
  • test crystal resonators 25, 25 ⁇ ' ⁇ 25 are arranged at the detection site of the flow path 22 of the line 1 and the flow path 22 of the line 1, while the line ⁇
  • the reference crystal unit 25 By disposing the reference crystal unit 25 in the flow path 22, it is possible to remove the influence of drift components on a plurality of types of measurement objects, for example, and to reduce measurement errors.
  • the present invention it is possible to provide a measuring instrument effective for measuring a measurement object contained in a sample to be analyzed such as a living body, food, and soil, a measurement method and a measurement apparatus using the measurement instrument. I'll do it.

Landscapes

  • Physics & Mathematics (AREA)
  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Analytical Chemistry (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • General Physics & Mathematics (AREA)
  • Immunology (AREA)
  • Pathology (AREA)
  • Apparatus Associated With Microorganisms And Enzymes (AREA)

Abstract

Lorsqu’un oscillateur piézoélectrique est agencé sur une partie de détection d’un récipient à réaction, on peut facilement générer un film de détection destiné à un oscillateur piézoélectrique et l’on peut facilement reproduire le capteur, permettant ainsi sa réutilisation. Un instrument de mesure (1) comprend un récipient à réaction (2) capable d’acheminer un objet de mesure (10) dans un échantillon. Le récipient à réaction (2) comprend un oscillateur piézoélectrique (6) agencé sur une partie de détection (5) capable de détecter un objet de mesure (10) ; un matériau générateur de force magnétique (7) capable de former de manière sélective et réversible un champ magnétique dans l’oscillateur piézoélectrique (6) ; et un porteur (11) formé à partir de particules magnétiques (11a) et d’un piégeur (11b) capable de lier spécifiquement l’objet de mesure (10), ou formé à partir de particules magnétiques (11a) et d’un objet (11c) semblable à l’objet de mesure et qui est transporté magnétiquement à la surface de l’oscillateur piézoélectrique (6) lorsqu’un champ magnétique est appliqué à l’oscillateur piézoélectrique (6). L’invention a également trait à un kit de mesure employant l’instrument de mesure (1), un procédé de mesure et un dispositif permettant de mesurer la concentration de l’objet de mesure, ainsi qu’un procédé permettant de reproduire un oscillateur piézoélectrique.
PCT/JP2006/315614 2005-08-05 2006-08-07 Instrument de mesure, kit de mesure employant cet instrument, procédé de mesure, dispositif de mesure, et procédé permettant de reproduire un oscillateur piézoélectrique WO2007018187A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
JP2005228921 2005-08-05
JP2005-228921 2005-08-05

Publications (1)

Publication Number Publication Date
WO2007018187A1 true WO2007018187A1 (fr) 2007-02-15

Family

ID=37727370

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/JP2006/315614 WO2007018187A1 (fr) 2005-08-05 2006-08-07 Instrument de mesure, kit de mesure employant cet instrument, procédé de mesure, dispositif de mesure, et procédé permettant de reproduire un oscillateur piézoélectrique

Country Status (1)

Country Link
WO (1) WO2007018187A1 (fr)

Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2011121859A1 (fr) * 2010-03-29 2011-10-06 独立行政法人科学技術振興機構 Élément de détection
DE102017006676A1 (de) * 2017-07-14 2019-01-17 Vdeh-Betriebsforschungsinstitut Gmbh Vorrichtung und Verfahren zur Bestimmung einer Konzentration von Partikeln in einem Fluid
US11391730B2 (en) * 2019-12-31 2022-07-19 Robert Bosch Gmbh Sensor refresh systems
CN116879105A (zh) * 2023-09-08 2023-10-13 云南绅博源生物科技有限公司 一种用于卷烟过滤材料制备的持水性检测方法及装置

Citations (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPS5954943A (ja) * 1982-09-22 1984-03-29 Daido Steel Co Ltd 流体中の磁性粒子量測定方法
JPS6435269A (en) * 1987-06-18 1989-02-06 Gene Trak Systems Signal amplification for measurement using piezo-electric oscillator
JPH04320967A (ja) * 1991-02-27 1992-11-11 Nec Corp 免疫センサ
JPH0688779A (ja) * 1992-09-07 1994-03-29 Terumo Corp 遺伝子センサー
JPH07225233A (ja) * 1994-02-15 1995-08-22 Kyowa Medex Co Ltd 水晶発振子を用いる免疫測定法
JP2000283905A (ja) * 1999-03-30 2000-10-13 Noboru Koyama マルチチャンネルｑｃｍセンサデバイス
JP2003307481A (ja) * 2002-04-17 2003-10-31 Canon Inc マルチチャネルバイオセンサ
WO2005031316A1 (fr) * 2003-09-29 2005-04-07 National Institute Of Advanced Industrial Science And Technology Procede et dispositif d'analyse automatique
JP2005274164A (ja) * 2004-03-23 2005-10-06 Citizen Watch Co Ltd バイオセンサー装置

Patent Citations (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPS5954943A (ja) * 1982-09-22 1984-03-29 Daido Steel Co Ltd 流体中の磁性粒子量測定方法
JPS6435269A (en) * 1987-06-18 1989-02-06 Gene Trak Systems Signal amplification for measurement using piezo-electric oscillator
JPH04320967A (ja) * 1991-02-27 1992-11-11 Nec Corp 免疫センサ
JPH0688779A (ja) * 1992-09-07 1994-03-29 Terumo Corp 遺伝子センサー
JPH07225233A (ja) * 1994-02-15 1995-08-22 Kyowa Medex Co Ltd 水晶発振子を用いる免疫測定法
JP2000283905A (ja) * 1999-03-30 2000-10-13 Noboru Koyama マルチチャンネルｑｃｍセンサデバイス
JP2003307481A (ja) * 2002-04-17 2003-10-31 Canon Inc マルチチャネルバイオセンサ
WO2005031316A1 (fr) * 2003-09-29 2005-04-07 National Institute Of Advanced Industrial Science And Technology Procede et dispositif d'analyse automatique
JP2005274164A (ja) * 2004-03-23 2005-10-06 Citizen Watch Co Ltd バイオセンサー装置

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
LI J. ET AL.: "[Piezoelectric immunosensor based on magnetic nanoparticles with simple immobilization procedures]", ANALYTICA CHIMICA ACTA, vol. 81, no. 2, 3 April 2003 (2003-04-03), pages 191 - 198, XP003003846 *

Cited By (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2011121859A1 (fr) * 2010-03-29 2011-10-06 独立行政法人科学技術振興機構 Élément de détection
JP4981998B2 (ja) * 2010-03-29 2012-07-25 独立行政法人科学技術振興機構 検出素子
US8438912B2 (en) 2010-03-29 2013-05-14 Japan Science And Technology Agency Detection device
DE102017006676A1 (de) * 2017-07-14 2019-01-17 Vdeh-Betriebsforschungsinstitut Gmbh Vorrichtung und Verfahren zur Bestimmung einer Konzentration von Partikeln in einem Fluid
WO2019012034A1 (fr) 2017-07-14 2019-01-17 Vdeh-Betriebsforschungsinstitut Gmbh Dispositif et procédé de détermination de la concentration de particules dans un fluide
DE102017006676B4 (de) 2017-07-14 2024-08-01 Vdeh-Betriebsforschungsinstitut Gmbh Vorrichtung und Verfahren zur Bestimmung einer Konzentration von Partikeln in einem Fluid
US11391730B2 (en) * 2019-12-31 2022-07-19 Robert Bosch Gmbh Sensor refresh systems
CN116879105A (zh) * 2023-09-08 2023-10-13 云南绅博源生物科技有限公司 一种用于卷烟过滤材料制备的持水性检测方法及装置
CN116879105B (zh) * 2023-09-08 2023-11-14 云南绅博源生物科技有限公司 一种用于卷烟过滤材料制备的持水性检测方法及装置

Similar Documents

Publication Publication Date Title
JP4587903B2 (ja) 測定器具及びこれを用いた測定用キット、測定方法並びに測定装置
US7389673B2 (en) Sensor for detecting analyte in liquid and device for detecting analyte in liquid using the same
US7749445B2 (en) Method and apparatus for analyzing bioprocess fluids
JP5922153B2 (ja) マイクロ流体アッセイ装置
US10466194B2 (en) Piezoelectric biochips having fluidic channels
US20080011058A1 (en) Piezoresistive cantilever based nanoflow and viscosity sensor for microchannels
WO2006103439A2 (fr) Cartouche pour analyseur d'echantillons fluides
JP2008502911A (ja) センサおよび較正要素のアレイを備えた分析機器
JP2008082988A (ja) 多段階増幅を利用した検出方法
JPH11183479A (ja) 溶液測定用センサ及び溶液成分測定方法
CN106546727B (zh) 一种石墨烯玻璃芯片的制备方法
WO2007018187A1 (fr) Instrument de mesure, kit de mesure employant cet instrument, procédé de mesure, dispositif de mesure, et procédé permettant de reproduire un oscillateur piézoélectrique
JP2005274164A (ja) バイオセンサー装置
EP1662257A1 (fr) Outil pour mesurer un objet a mesurer, dispositif de mesure et procede de mesure
WO2016035197A1 (fr) Cartouche pour capteur électrochimique de l'immunité et dispositif de mesure l'utilisant
US8250908B2 (en) Wave sensor apparatus including gas removing unit and method of detecting target material in liquid sample
JP5240794B2 (ja) 感知装置
JP2007093239A (ja) Ｑｃｍ分析装置
JP4127192B2 (ja) 水晶振動子電極表面へのレセプタ固定方法およびレセプタ固定水晶振動子の製造方法
EP2284539B1 (fr) Méthode et appareil pour analyser les fluides bioprocédés
JP2007085973A (ja) Ｑｃｍ分析装置
da Silva Granja et al. Characterisation of particle-surface interactions via anharmonic acoustic transduction
JP2690849B2 (ja) 水晶振動子式バイオセンサの製法及び水晶振動子式バイオセンサ
Kato et al. 1Pa-28 Development of a High-Frequency Electrodeless Quartz Crystal Microbalance Chip with a Bare Quartz Resonator Encapsulated in a Silicon MicroChannel and Its Application to a Biosensor
JP5040019B2 (ja) マイクロリアクター及びマイクロリアクターシステム

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application
NENP Non-entry into the national phase

Ref country code: DE

122 Ep: pct application non-entry in european phase

Ref document number: 06782456

Country of ref document: EP

Kind code of ref document: A1

NENP Non-entry into the national phase

Ref country code: JP