JP2005204661A - Detecting apparatus for molecule derived from organism, dioxins and endocrine disrupter, and method for detection using the same - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a detecting apparatus for a molecule derived from an organism, dioxins, and an endocrine disrupter capable of easily realizing detection of the molecule derived from the organism, dioxins, and the endocrine disrupter by a rapid and stepwise operation of the detecting apparatus, and to provide a configuration of the method for the detection. <P>SOLUTION: This invention relates to the detecting apparatus for the molecule derived from the organism, the dioxins and the endocrine disrupter, which carries out the detection after subjecting these samples to a pretreatment, wherein the apparatus is provided with a chip having in its inside at least one treating vessel for performing the pretreatment and a detecting vessel (17) installed downstream of the treating vessel to detect a prescribed molecule derived from the organism, the dioxins and the endocrine disrupter, and at least a melting/softening valve (11) installed on the boundary of the detecting vessel (17) and the treating vessel to make the molecule derived from the organism, the dioxins and the endocrine disrupter melt or soften and pass by heating or irradiating with an ultrasonic wave. The method for detecting them by using the apparatus is also provided. <P>COPYRIGHT: (C)2005,JPO&NCIPI

Description

  The present invention is artificially generated through industrial activities such as biological molecules derived from nucleic acid such as DNA and RNA, protein structural units such as peptides and sugar chains, and dioxins and endocrine disrupting substances (environmental hormones, environmental estrogens, etc.). The present invention relates to a detection device for detecting a harmful substance.

  Recently, serious damage has been caused by infectious diseases such as bovine spongiform encephalopathy (BSE) and severe acute respiratory syndrome (SARS). As a measure to prevent the spread of viruses, samples related to biological molecules are collected from livestock suspected of being infected, and the samples are reacted by the PCR method, then processed by electrophoresis, etc. for analysis and detection of viruses. Surveys are being conducted.

  In the field of DNA detection, devices that perform high-speed separation of DNA in a very small amount and devices using a micro-channel that stabilizes detection accuracy have already been developed.

  For example, Patent Document 1 describes a technique for analyzing DNA or the like using a sample by capillary gel electrophoresis.

  Patent Document 2 describes a technique for efficiently passing a working fluid through a microchannel.

  Further, Patent Document 3 describes a technique for passing a fluid using a wax valve.

Patent Document 4 describes a technique in which a valve is provided in a micro flow path, the valve is opened and closed by air pressure, and a working fluid is allowed to pass through.
JP-A-6-74936. JP 2001-252896 A. JP2003-270252A. JP 2004-33919 A.

  However, these previous surveys or inspections are conducted by specialists in laboratories equipped with facilities and environments, so that it is possible to avoid the tendency of taking time to understand the results and increasing the cost. It was impossible.

  In addition, these are biologically derived molecules such as DNA, and dioxins, and further artificially generated harmful substances such as endocrine disrupting substances (hereinafter, the biologically derived molecules and the harmful substances are collectively referred to as “detection objects”). )), It is difficult to carry out the separation and detection in a continuous manner, and since it is almost impossible to transport as a device, there are problems that a special place is required to operate the device. ing.

  Due to this situation, in the case of each of the above-mentioned devices, in order to obtain a test result after immediately collecting and detecting a sample including a detection target (hereinafter referred to as “sample”), We cannot escape the evaluation that it is insufficient.

  Under such circumstances, there is a need for an inexpensive analytical instrument or analysis chip that can collect a sample, and can promptly investigate or inspect it, and can obtain a result immediately.

  The present invention relates to a detection target object detection device capable of quickly detecting a detection target object by a simple operation in one detection device, and a configuration related to a detection target detection method using the device. It is an issue to provide.

In order to solve the above problems, the basic configuration of the present invention is as follows.
(1) An apparatus for detecting a detection object from a sample after pre-processing the sample,
In the chip, provided with at least one or more processing tanks for pre-processing the sample, and a detection tank provided downstream of the processing tank for detecting the detection object,
At least at the boundary between the detection tank and the processing tank, a melt-softening valve that can be dissolved in the sample containing the detection target and allowed to pass the sample containing the detection target by melting or softening was provided. A device for detecting a detection object based on
(2) A method of detecting a detection object from a sample after pre-processing the sample,
After performing an operation for pre-processing the sample in the processing tank, an operation for detecting the detection target contained in the sample is performed in the detection tank,
At the boundary between the detection tank and the processing tank, a melt-softening valve that melts or softens due to heating or applying ultrasonic waves is provided,
After the operation in the processing tank is completed, an external force is applied to the sample or the processing tank containing the sample, and the melting / softening valve is heated or ultrasonically applied to open the detection target. A method for detecting an object to be detected, the method comprising: moving a containing sample to a detection tank;
Consists of.

  The configuration of the method (2) is based on the configuration of the device (1), and is characterized by melting or softening the melt-softening valve by heating or applying ultrasonic waves. Yes.

  The treatment tanks in the constitutions (1) and (2) are, for example, a tank that performs a process for separating insoluble polymers from low molecules, a tank that performs a process for removing insoluble impurities, and a reaction such as amplification of DNA. It is a tank that performs chemical and biochemical reaction operations such as a tank that performs the treatment.

  In the configurations of (1) and (2), at least one processing tank that pre-processes the sample is provided in the chip, and is provided downstream of the processing tank to detect the detection target. A detection tank, and at least a boundary between the detection tank and the processing tank is provided with a valve through which the sample can be passed by melting or softening. The sample can be retained in the processing tank by moving it to the processing tank and closing the valve.

  Further, unlike the conventional mechanical valve, the inside of the tank is completely sealed, and each processing tank partitioned by the valve can be separated chemically and biochemically.

  Furthermore, since the melted valve dissolves in the sample and its components are diluted, it cannot harden again and block the subsequent flow path or valve.

  In the present invention, by opening and closing the valve between the treatment tanks, the treatment in each treatment tank can be performed in a cascade manner, so that individual reaction conditions (for example, temperature, time, etc.) can be precisely controlled. I can do it.

  In addition, when a centrifugal force is employed as a means for moving the sample, only a power supply and a table centrifuge are sufficient as support devices other than the chip.

  Therefore, in the present invention, it is possible to quickly detect the detection target from the collected sample, and since the configuration of the detection apparatus is simple, its operation is extremely easy, and the conventional analysis apparatus Compared to the above, it is possible to obtain an advantage that the economic cost is low.

As embodiment regarding the apparatus of said (1),
(A) A mechanism for releasing the air to be excluded in the adjacent downstream processing tank or detection tank when the sample and the valve dissolved in the sample move from the upstream processing tank to the adjacent downstream processing tank or detection tank (B) A configuration characterized in that at least one of the treatment tanks is an insoluble matter filter tank for removing insoluble contaminants,
(C) A reaction tank which is one of the treatment tanks is provided between the insoluble matter filter tank and the detection tank, and the valve is provided at the boundary between the reaction tank and the detection tank. Constitution,
(D) The insoluble matter filter tank is set in a plurality of stages, and the valve is provided on all or part of the boundary for each stage,
(E) a configuration in which the reaction tank is set in a plurality of stages, and the valve is provided on all or part of the boundary of each stage;
(F) In the detection tank, any one of the electrophoresis method, the light detection method, the evanescence method, and the NMR method is adopted,
(G) In the detection tank, after adopting the electrophoresis method, a buffer solution holding tank is provided around the detection tank, and the buffer solution is melted or softened between the buffer solution holding tank and the detection tank. A configuration provided with a valve that can be moved between the buffer solution holding tank and the detection tank;
(H) A configuration characterized by providing a valve that melts or softens due to heating,
(I) A structure characterized in that a heating electrode is installed in the vicinity of these valves as a heating mechanism for the valves;
(J) Adopting a mixture of polysaccharides such as agarose, chitosan, gellan gum, pullulan, carrageenan, xyloglucan, dextran, dextrin, cellulose gum, amylose, amylopeptine, etc. Features configuration,
(K) A configuration characterized by providing a valve that melts or softens due to application of ultrasonic waves,
Can be adopted.

  Also in the configuration of the method (2), the (a), (b), (c), (d), (e), (f), (g), (h), (i), (j ) And (k) can be employed (however, the description of these embodiments is only a reproduction of the above (a) to (k), and the description is omitted). I will do it.)

  Hereinafter, specific description will be given with reference to the drawings.

  FIG. 1 shows an embodiment of a detection object detection apparatus according to the present invention.

  The melt-softenable valve (11) is conducting current from surrounding electrodes (not shown in FIG. 1 but shown by 32 in FIG. 2 and 52 in FIG. 3). A material that is melted or softened by electric heat in each conductive portion (21) and can be made into a state that does not cause any trouble in the analysis of the detection target is adopted.

In the embodiment shown in FIG. 1, the heating uses electric heat generated in each conduction part (21). However, the heating to the melt-softening valve (11) is not limited to the method using conduction, for example, There is also a technique in which a needle-like heating element is brought close to the melt-softening valve (11) to generate heat locally.
However, in order to perform stable heating, it is more convenient to heat the conductive portions (21) based on conduction of current from the electrodes.

  In the embodiment shown in FIG. 1, the insoluble matter filter tank (12) has a sample injection window (13) and an insoluble matter removing filter (14) installed in two stages for first removing insoluble impurities. ), A three-stage configuration is adopted in which a microfilter (15) capable of removing fine insoluble impurities is provided.

  With such a three-stage configuration, it is possible to increase the filter removal efficiency of insoluble contaminants and increase the purity of the detection target in the sample, but the number of stages of the insoluble filter tank (12) is four. It is of course possible to set more than one stage if necessary.

The gel filtration tank (18) is installed for the purpose of separating the insoluble polymer from the low molecule.
The gel filtration tank (18) further includes a second microfilter (19) for removing fine insoluble impurities.

  The reaction tank (16) is interposed between the insoluble matter filter tank (12) and the detection tank (17) employing the electrophoresis method, and can be melted and softened between the detection tank (17). A valve (11) is provided.

  In the reaction tank (16), for example, a PCR reaction for amplifying DNA employed in DNA detection, a cell proliferation reaction, and the like can be performed.

  The detection tank (17) is connected to the reaction tank (16) via the melt-softening valve (11), and in the case of the embodiment according to FIG. 1, detection is performed by electrophoresis. Therefore, a buffer solution holding tank (22) is provided in the periphery, and a melt-softening valve (11) is also provided between the reaction tank (16) and the buffer solution holding tank (22). Yes.

  The buffer solution holding tank (22) is a tank for holding a buffer solution for electrophoresis until the time of electrophoresis.

  When the electrophoresis method is adopted in the detection tank (17), it is indispensable to provide the electrode for electrophoresis and the conduction part (23).

  As described above, the detection tank (17) can adopt not only the electrophoresis method but also, for example, a light detection method, an evanescence method, an NMR method, etc. In this case, a buffer solution holding tank ( 22) is not always necessary.

  FIG. 2 is a perspective view of an embodiment relating to a test pattern of a valve that can be opened by melting or softening due to heating in the present detection device, FIG. 3 is a cross-sectional view, and FIG. It is the perspective view which showed the test pattern of the board | substrate which performs a movement with a centrifugal force.

  FIG. 5 is a cross-sectional view showing an example of the substrate manufacturing method.

  When producing this detection apparatus, as shown in FIG. 5, the base material (71) used as a base is prepared first. The base material (71) is a material having no brittleness, for example, a plastic such as silicon resin (polydimethylsiloxane (PDMS), acrylic resin, vinyl chloride, PTFE, polycarbonate resin, polyethylene terephthalate (PET), metal, glass, etc. A mixture of ceramic, etc., or a combination thereof can be used as the material.

  A microchannel forming part (72) is pasted and formed on the substrate (71). The micro-channel may be formed by etching technique or hot press using a mold. In addition, any of molding by laser processing, molding using a micromachining process, and the like can be employed.

  Next, an insoluble matter removing filter (14) and a microfilter (15) are formed in a region where the insoluble matter filter tank (12) is formed in the microchannel forming portion (72), and further, a second microfilter ( 19).

  The material of the filter can be typically used as a dry film, resin, metal, glass, ceramic, etc., but is not limited thereto, and is not dissolved in the sample and has a filtration function, and Any material that can withstand centrifugal force and the like may be used.

  Examples of processing methods include laser processing and etching processing in the case of dry film, etching processing in the case of resin, laser processing, press processing, micromachining process, etc., in the case of metal, plating processing, etching processing, laser processing, In the case of glass, ceramic, etc., an etching process, a laser process, a press process, a micromachine process, etc. are mentioned, such as a press process and a micromachining process.

  The filter shape is, for example, a slit shape as shown in FIG. 1, base material (base) (71) and lid (73)? A shape in which a plurality of columns are alternately shifted between the two is employed, but in short, it is necessary to have a shape that can filter insoluble impurities.

  After attaching the microchannel forming part (72), as shown in FIGS. 2 and 3, the valve in the microchannel forming part (72) can be melted or softened due to heating. (Hereinafter, abbreviated as “valve melting point”.) 31 and 51 are formed with electrodes 32 and 52 for applying heat.

As an electrode forming method, any of a conductive tape, a metal foil, a metal piece at a predetermined location, metal plating, metal vapor deposition, sputtering, and the like can be employed.
In any case, the metal used is a conductive metal such as copper.

  When the melt-softening valve (11) is installed in the micro-channel forming the micro-channel forming part (72), the melt-softening in the melted or softened state at a high temperature at the valve melting point (31, 51). The valve can be formed by injecting the material of the possible valve (11) and curing by allowing to cool (the injection is usually performed by a microsyringe in many cases).

  The valve melting points (31, 51) are formed by the same method as that for the microchannel when the microchannel is formed.

  In addition, the shape of the valve melting point (31, 51) in the microchannel is rectangular in the test pattern shown in FIG. 2, but is not limited to this, and is in a gel form before melting or softening. The valve material can be retained and can be melted or softened, or can be passed through the valve material that is in the form of a sol in the melting or softening process. For example, as shown in FIGS. Such a square, circle, rhombus, hexagon, home base, triangle, ellipse, oval, etc.

  A typical example of the material used for the valve is an agarose gel, but the material is not limited to an agarose gel, and an aqueous gel (nanogel) containing a gelling agent and having a volume of 1 microliter or less. Any material that melts or softens in accordance with the sol-gel transition and opens based on the melting or softening, such as chitosan gel, gellan gum gel, pullulan gel, carrageenan gel, xyloglucan gel, A mixture of polysaccharides such as dextran gel, dextrin gel, cellulose gum gel, amylose gel, and amylopectin gel or a combination thereof can also be sufficiently employed because it does not affect the detection of the detection target.

  As a method for injecting the gel, in addition to a manual method such as a microsyringe, an automatic method such as an ink jet, a spotter, or a micropump can be employed.

As a lid for the base material (71), silicon resin (PDMS, etc.), acrylic resin, vinyl chloride, polycarbonate resin, plastic such as PET, PTFE, metal, glass, ceramic, etc., or a mixture thereof is pasted with an adhesive. Although it can be formed by attaching, it is better to be transparent in order to observe the sample.
Alternatively, the microchannel may be formed by directly processing the substrate (71) without attaching the microchannel forming portion (72) to the substrate (71). In this case, the same processing method as described above is used.

  A predetermined reagent is injected into a region of the base material (71) where the detection tank (17) is formed. As the reagent injection method, a manual method such as a microsyringe, an inkjet, a spotter, a micropump, or the like is used. It is possible to adopt the automatic method.

  In order to confirm the function of the melt softening valve (11) to block the flow of the sample, the sample is injected in the upstream processing tank, the acceleration is applied to the chip, and the acceleration is applied to the chip. It is good to confirm that the sample moves from the upstream tank to the adjacent downstream processing tank due to inertia, and that the flow of the sample is stopped at the valve section.

  In order to confirm that the valve melts or softens due to heating, the tip of the valve is melted or softened by applying voltage to the electrode part and heating the valve part. (For example, when a mechanism for applying a centrifugal force is applied, a centrifugal force is applied by rotation), and it is good to confirm that the sample passes with the material constituting the melted or softened valve. .

In order to confirm the filter function in the insoluble matter filter tank (12), the sample is set in the Eppendorf tube from the resin (73) provided with the sample inlet to the insoluble substance filter tank (12). It is better to confirm that the insoluble contaminants can be removed by applying acceleration to the Eppendorf tube.
As the mechanism for moving the sample, a mechanism that uses the inertia of the sample or a mechanism that applies pressure to the sample can be employed. The operation using inertia does not require an external precision mechanism for controlling pressure and flow velocity, and the sample can be moved at a low cost and in the most stable state.

  In addition, as a mechanism that uses the inertia of the sample, a mechanism that fixes the chip on the linear feeder and vibrates the chip may be used.

  In addition, as a mechanism for applying pressure to the sample, an operation using a piston, a cylinder, or the like and pressing the sample with a pressure of a fluid (for example, air) that does not mix with the sample can be employed.

  In FIG. 7 (a), the gel-like melt-softening valve (11a) is solated as shown in the melt-softening valve (11b) of FIG. 7 (b) by applying heat or ultrasonic waves. The solubilized melt-softening valve (11b) is dissolved in the sample (83), and the sample (85) including the dissolved valve is air (84) present in the downstream processing tank (82) by centrifugal force. ) And move from the upstream processing tank (81) to the downstream processing tank (82). At that time, the excluded air (84) moves to the upstream processing tank (81) through the exclusion passage (10).

  Hereinafter, DNA is selected as the detection target, agarose is adopted as the material constituting the valve, and a mechanism for applying acceleration to the chip by centrifugal force is adopted as the mechanism for moving the sample. To explain.

  A commercially available acrylic resin having a thickness of 3 mm was prepared. This acrylic resin was dry-etched using a carbon dioxide laser under the conditions of an output of 25 W, a laser diameter of 70 microns, and a wavelength of 10.6 microns to form a microchannel having a channel depth of 1 mm.

  The formed channel was washed with ethanol and dried.

  A commercially available copper foil (0.018 mm thickness) was attached to a predetermined portion of the formed flow path with an instantaneous adhesive to form an electrode.

  A pouch film (0.1 mm thick) was attached to the substrate by thermocompression bonding to form a test pattern.

  A 2% by weight aqueous solution of agarose prepared at 66 ° C. was injected into a valve portion formed by the same method as that for the microchannel, using a microsyringe. After naturally cooling for 10 minutes, it was confirmed that the temperature reached room temperature (24 ° C.), and the formation of the valve portion (gelation of 2% by weight aqueous solution of agarose) was completed.

  This substrate was fitted into an Eppendorf tube. The Eppendorf tube containing the substrate was centrifuged at 9000 rpm for 5 seconds.

  Even after centrifugation, the gelled agarose was retained. A sample was injected into this substrate with a microsyringe, and this substrate was fitted into an Eppendorf tube.

  The Eppendorf tube containing the substrate was centrifuged at 6000 rpm for 5 seconds.

  Observation after the centrifugation operation confirmed that the sample flowed through the flow path. Moreover, the sample was hold | maintained before the agarose gel valve, and it has confirmed that the agarose gel fully fulfill | performed the role of the valve.

  The base material was taken out from the Eppendorf tube, and a current of 1.7 A was applied to the electrode for 30 seconds to warm the gel part to 66 ° C. At this time, it was confirmed that the agarose gel was a fluid.

  After heating, the tube was fitted again into an Eppendorf tube, and centrifuged again at 6000 rpm for 5 seconds.

  After observing the centrifugation, it was confirmed that the sample and the solated agarose gel were mixed, passed through the valve portion and the flow path, and all flowed down to the bottom of the Eppendorf tube.

  This confirmed that an agarose gel can be used as a valve.

  A commercially available acrylic resin substrate having a thickness of 1 mm was prepared. This acrylic resin was degreased and washed with a 5% aqueous alkali solution at 50 ° C. for 5 minutes, washed with pure water, neutralized with a 5% aqueous sulfuric acid solution, further washed with pure water, and dried.

  An acrylic dry film resist (0.025 mm thick) was laminated on the acrylic resin substrate by thermocompression bonding, and then the dry film resist was exposed according to a predetermined photomask pattern (60 mJ / cm 2), and further 1% at 30 ° C. -Development with an alkaline aqueous solution and patterning were performed to obtain a flow path circuit.

  The substrate was sufficiently washed with pure water and then dried. A pouch film (thickness: 0.1 mm) provided with a window serving as a sample injection port was attached to the substrate by thermocompression bonding to form a test pattern.

  This substrate was cut according to the inner diameter of the Eppendorf tube. After cutting, a sample was injected from the sample injection port, and this substrate was fitted into an Eppendorf tube.

  The Eppendorf tube containing the substrate was centrifuged at 6000 rpm for 3 seconds.

  When the centrifugation operation was observed, the sample passed through the channel and reached the lower part of the Eppendorf tube.

  From this result, it was confirmed that the produced substrate can sufficiently move the fluid using the centrifugal force.

  A commercially available acrylic resin having a thickness of 3 mm was prepared. This acrylic resin was dry-etched using a carbon dioxide laser under the conditions of an output of 25 W, a laser diameter of 70 microns, and a wavelength of 10.6 microns to form a microchannel having a channel depth of 1 mm.

  The formed channel was washed with ethanol and dried.

  A pouch film (thickness: 0.1 mm) provided with a window serving as a sample injection port was attached to the substrate by thermocompression bonding to form a test pattern.

  A sample was injected into the substrate with a microsyringe, and this substrate was fitted into an Eppendorf tube.

  The Eppendorf tube containing the substrate was centrifuged at 12000 rpm for 5 seconds.

  Observation after the centrifugation operation confirmed that the sample flowed through the flow path as described above.

  From the above, it was confirmed that a small amount of sample in the microchannel can be moved using centrifugal force.

  A commercially available acrylic resin substrate having a thickness of 1 mm was prepared. This acrylic resin was degreased and washed with a 5% aqueous alkali solution at 50 ° C. for 5 minutes, washed with pure water, neutralized with a 5% aqueous sulfuric acid solution, further washed with pure water, and dried.

  After laminating an acrylic dry film resist (0.075 mm thickness) on the acrylic resin substrate by thermocompression bonding, the dry film resist was exposed according to a predetermined photomask pattern with a spacing of 0.04 mm (70 mJ / cm 2). Further, development was performed with a 1% -alkali aqueous solution at 30 ° C. and patterning was performed.

  At this time, a mortar-shaped pattern having a gap of 0.04 mm and a depth of 0.03 mm was obtained.

  The substrate was sufficiently washed with pure water and dried, and then an acrylic dry film resist (thickness: 0.015 mm) was laminated on the substrate again by thermocompression bonding. This dry film resist was alignment-exposed with the photomask (60 mJ / cm 2), and further developed with a 1% -alkaline aqueous solution at 30 ° C. and patterned. At this time, a mortar-shaped pattern having a gap of 0.04 mm and a depth of 0.045 mm was obtained.

  The substrate was sufficiently washed with pure water and then dried. A pouch film (thickness: 0.1 mm) provided with a window serving as a sample injection port was attached to the substrate by thermocompression bonding to form a test pattern.

  This substrate was cut according to the inner diameter of the Eppendorf tube. After cutting, a plant cell extract (tomato juice drink) was injected from the sample injection port, and this substrate was fitted into an Eppendorf tube.

  The Eppendorf tube containing the substrate was centrifuged at 6000 rpm for 3 seconds.

  On the other hand, as a control, a similar sample was attached to the upper part of the inner wall of the Eppendorf tube without a microchannel substrate, and a centrifugal separation operation was similarly performed at 6000 rpm for 3 seconds.

  Observation after the centrifugation showed that most of the insoluble components in the sample were adsorbed due to non-specific binding between the microchannel substrate, but only the highly soluble components passed through the channel and passed through the bottom of the Eppendorf tube. Reached. In contrast, in the control experiment, all components reached the bottom, and the same color (dark red) as the original was confirmed.

  From this result, it was found that the produced substrate can sufficiently satisfy the filter function.

[Comparative Example 1]
The probe DNA was attached to a commercially available slide glass for DNA chip with a spotter. On the other hand, the detection target was subjected to gene amplification using a PCR device, and a fluorescent substance (CY3) was added to the amplified detection target. This detection object was dropped on the adjusted DNA chip, and hybridization was performed. After hybridization, DNA was detected with a scan array light. As described above, according to the prior art, various expensive specialized equipments such as a device for spotting DNA on a chip, a device for amplifying DNA, a hybridization device, a chip reading device (scanner) are required, and each process is further performed. Turned out to be very skillful.

  The detection device according to the present invention can be used in the livestock field, the medical field, the biology research field, and the manufacturing field related to these fields.

It is sectional drawing of the longitudinal direction which shows the best embodiment in this invention. It is the perspective view which showed the test pattern of the board | substrate which a valve opens with heat. It is sectional drawing of the direction orthogonal to the longitudinal direction which shows the test pattern of the board | substrate which a valve opens with heat. It is the perspective view which showed the test pattern of the board | substrate which performs a fluid movement with a centrifugal force. FIG. 4 is a cross-sectional view showing a process of preparing a base material, a process of forming a micro flow path forming part for performing filtering in an insoluble matter filter tank, and further affixing a resin provided with a sample inlet. . It is sectional drawing of the moving direction in a valve melting location, (a) shows the case of a circle, (b) shows the case of a rhombus, (c) shows the case of a hexagon, (d) shows a home base shape (E) shows the case of a triangle, (f) shows the case of an oval, (g) shows the case of a shape in which each apex angle of the triangle has a large R, ( h) shows a case where the hexagon of (c) is rotated by 90 °. In this invention, it is sectional drawing of the moving direction of a sample which shows the process in which a sample moves to the downstream downstream processing tank from an upstream processing tank, (a) shows the state before a valve softens, b) shows a state immediately after the valve is softened, and (c) shows that the softened valve is dissolved in the sample, and then an external force is applied to the sample or the detection device, and the sample containing the dissolved valve is in the upstream processing tank. (D) shows a state in which the movement of the sample including the dissolved valve from the upstream processing tank to the adjacent downstream processing tank has been completed.

Explanation of symbols

DESCRIPTION OF SYMBOLS 10 Exclusion passage 11 Melt softening possible valve 12 Insoluble matter filter tank 13 Sample injection window 14 Insoluble matter removal filter 15 Micro filter 16 Reaction tank 17 Detection tank 18 Gel filtration (SEC) tank 19 Second micro filter 21 Current flows from the heating electrode Conduction portion 22 to be conducted Buffer holding tank 23 Electrode for electrophoresis and conduction portion 31 Valve melting point (however, as viewed from above in the flow path direction)
32 Heating electrode 41 Sample inlet 51 Valve melting point (however, according to a side view perpendicular to the flow path)
52 Heating electrode 53 Resin 71 with adhesive 71 Base material (base)
72 Micro-channel forming part 73 Lid 81 Upstream processing tank 82 Downstream processing tank 83 Sample 84 containing detection object Air 85 Sample containing dissolved valve and detection object

Claims (15)

An apparatus for detecting biologically derived molecules, dioxins, endocrine disrupting substances from the sample after pre-processing the sample,
In the chip, comprising at least one or more treatment tanks for pretreatment of the sample, and a detection tank provided downstream of the treatment tank for detecting the biological molecules, dioxins, and endocrine disrupting substances,
At least at the boundary between the detection tank and the treatment tank, it is dissolved or dissolved in a sample containing biologically derived molecules, dioxins and endocrine disrupting substances, and contains biologically derived molecules, dioxins and endocrine disrupting substances. An apparatus for detecting biologically derived molecules, dioxins, and endocrine disrupting substances based on the provision of a melt-softening valve through which a sample can pass.   When a sample containing a biological molecule, dioxins, endocrine disrupting substance and a melt-softening valve dissolved in the sample move from an upstream processing tank to a downstream processing tank or detection tank adjacent to the upstream processing tank, 2. The apparatus for detecting biologically derived molecules, dioxins and endocrine disrupting substances according to claim 1, further comprising a mechanism for escaping air to be excluded from the downstream processing tank or detection tank.   The apparatus for detecting biologically derived molecules, dioxins and endocrine disrupting substances according to claim 1 or 2, wherein at least one of the treatment tanks is an insoluble matter filter tank for removing insoluble contaminants. A reaction tank which is one of the treatment tanks is provided between the insoluble matter filter tank and the detection tank, and the melt-softening valve is provided at the boundary between the reaction tank and the detection tank. The apparatus which detects the biological origin molecule | numerator, dioxins, and endocrine disrupting substance of Claim 3. The biologically derived molecule or dioxin according to claim 3 or 4, wherein the insoluble matter filter tank is set in a plurality of stages, and the melt-softening valve is provided in all or part of the boundary part of each stage. A device that detects endocrine disruptors. 5. The bio-derived molecule, dioxins, and endocrine according to claim 4, wherein the reaction tank is set in a plurality of stages, and the melt-softening valve is provided in all or a part of the boundary for each stage. A device that detects disturbing substances. The insoluble matter filter tank is set in a plurality of stages, and the melt-softening valve is provided in all or a part of the boundary part for each stage, biomolecules according to claim 6, dioxins, A device that detects endocrine disruptors.   In the detection tank, any one of an electrophoresis method, a light detection method, an evanescence method, and an NMR method is adopted, The biological molecule, dioxins, and endocrine disruption according to any one of claims 1 to 7 A device that detects substances. In the detection tank, after employing the electrophoresis method, a buffer solution holding tank is provided around the detection tank, and the buffer solution is melted or softened between the buffer solution holding tank and the detection tank. The biomolecule, dioxins, and endocrine disrupting substances according to any one of claims 1 to 7, wherein a melt-softening valve that can be moved between the holding tank and the detection tank is provided. Device to detect. An apparatus for detecting biomolecules, dioxins, and endocrine disrupting substances according to any one of claims 1 to 9, wherein a melt-softening valve that melts or softens due to heating is provided. 11. The apparatus for detecting biologically derived molecules, dioxins and endocrine disrupting substances according to claim 10, wherein a heating electrode is provided in the vicinity of these melt softening valves as a heating mechanism for the melt softening valves. As a material for the melt-softening valve, it is characterized by adopting a mixture of polysaccharides such as agarose, chitosan, gellan gum, pullulan, carrageenan, xyloglucan, dextran, dextrin, cellulose gum, amylose, amylopectin, or a combination thereof. The apparatus which detects the biological origin molecule | numerator, dioxins, and endocrine disrupting substance of Claim 10 or 11. The biomolecule, dioxins, and endocrine disrupting substances according to any one of claims 1 to 9, wherein a melt-softening valve that melts or softens due to application of ultrasonic waves is provided. apparatus. As a material for the melt-softening valve, it is characterized by adopting a mixture of polysaccharides such as agarose, chitosan, gellan gum, pullulan, carrageenan, xyloglucan, dextran, dextrin, cellulose gum, amylose, amylopectin, or a combination thereof. The apparatus which detects the biological origin molecule | numerator, dioxins, and endocrine disrupting substance of Claim 13. A method for detecting biologically derived molecules, dioxins, endocrine disrupting substances from the sample after pre-treatment of the sample,
After performing an operation for pretreatment of the sample in the treatment tank, an operation for detecting biologically derived molecules, dioxins, and endocrine disrupting substances contained in the sample is performed in the detection tank.
At the boundary between the detection tank and the processing tank, a melt-softening valve that melts or softens due to heating or applying ultrasonic waves is provided,
After the operation in the treatment tank is completed, an external force is applied to the sample or the treatment tank containing the sample, and the melt-softening valve is heated or opened by applying an ultrasonic wave, A method for detecting biologically derived molecules, dioxins, and endocrine disrupting substances, wherein a sample containing dioxins and endocrine disrupting substances is moved to a detection tank.

Images