JP2006284323A - Micro total analysis system - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a micro total analysis system capable of safely performing a desired inspection without leaking waste fluid such as a specimen or the like stored in a waste fluid reservoir disposed on the bottom of the inspection chip. <P>SOLUTION: In the micro total analysis system, the inspection chip has the waste fluid reservoir for reserving waste fluid produced in the reaction between an organism material and the reagent and its detection, and a water absorption body for holding the waste fluid is disposed in the waste fluid reservoir. Thus, the desired inspection can be safely performed without leaking the waste fluid such as the specimen stored in the waste fluid reservoir disposed on the bottom of the inspection chip. <P>COPYRIGHT: (C)2007,JPO&INPIT

Description

  In the present invention, a test chip on which a series of micro flow paths for performing mixing, reaction, and detection of a biological substance and a reagent contained in a specimen is formed is attached to a system body, and the test is performed by a micro pump unit. The present invention relates to a micro total analysis system that automatically performs mixing, reaction, and detection of a reaction between a biological substance and a reagent in a test chip while feeding a liquid in a fine channel of the chip.

  In recent years, by making full use of micromachine technology and ultrafine processing technology, devices and means (for example, pumps, valves, flow paths, sensors, etc.) for performing conventional sample preparation, chemical analysis, chemical synthesis, etc. have been miniaturized. Systems integrated on a chip have been developed.

  This is also called μ-TAS (Micro Total Analysis System), bioreactor, Lab-on-chips, biochip, and is used in the medical examination / diagnosis field, environmental measurement field, and agricultural production field. Application is expected.

  In particular, as seen in genetic testing, when a complicated process, skilled technique, and operation of equipment are required, the microreactor as an automated, high-speed and simplified micro-analysis system is cost-effective. The effect of enabling analysis not only in the required sample amount and the required time but also in any time and place is enormous.

  In the field where various tests such as clinical tests are performed in this way, quantitativeness, accuracy of analysis, and the like are regarded as important in measurement using a chip-type microreactor that produces results quickly regardless of location.

In addition, since the microreactor has severe restrictions in terms of its size and form, it is a problem to establish a liquid delivery system having a simple configuration and high reliability.
The present inventors have already proposed a micro total analysis system suitable for gene amplification reaction and its detection (Patent Documents 1 to 6).

The present inventors have already proposed a micropump system suitable for a microfluidic control element having high accuracy and excellent reliability (Patent Documents 7 and 8).
In such a micro comprehensive analysis system, a liquid such as a reagent in a fine flow path of a test chip is fed by a micropump by taking a driving liquid into the test chip.

  Specifically, a micro pump unit in which a plurality of piezo pumps using a piezo element having the configuration disclosed in Patent Documents 7 and 8 are provided on a chip and a test chip, for example, the flow passage openings of each other match. As described above, the chip-side surface of the micropump unit and the chip surface of the inspection chip are brought into close contact with each other so that the flow path on the pump side and the flow path on the inspection chip side are communicated.

Further, the driving liquid can be fed to the inspection chip by the action of the actuator connected to the pump.
Japanese Patent Application No. 2004-138959 Japanese Patent Application No. 2004-169912 Japanese Patent Application No. 2004-310744 Japanese Patent Application No. 2004-31313 Japanese Patent Application No. 2004-31314 Japanese Patent Application No. 2004-31315 JP 2001-322099 A JP 2004-108285 A

  By the way, the test chip used in such a micro total analysis system has a sample volume that can be introduced into the test chip has an upper limit in relation to the volume, so such sample volume may not be within the measurable range. An operation for preliminary concentration or separation of a specimen, or a mechanism capable of easily detecting and quantifying a minute amount of a reaction product with high sensitivity is required to be mounted on a test chip.

In particular, when a biological fluid such as blood is used as a specimen, it cannot often be used as it is for analysis, and some kind of pretreatment is usually added.
In this case, the liquid or specimen used for the pretreatment becomes unnecessary after the processing, and is stored in the waste liquid storage section provided at the bottom of the test chip, and the test chip that has finished the desired test is stored in the waste liquid storage section. Usually, the waste liquid is collected and incinerated while being stored.

  However, such a test chip has an airtightness in the waste liquid storage part, so there is little concern about waste liquid leakage under normal use conditions, but in the unlikely event of a malfunction in the test chip, the system body This may cause leakage of waste liquid.

In particular, when collecting and disposing of the inspection chip, it is likely to be complicated and the risk of leakage of waste liquid from the inspection chip becomes higher.
In view of such a current situation, the present invention is a micro total analysis system capable of performing a desired test safely without leakage of a waste liquid such as a specimen stored in a waste liquid storage part provided at the bottom of a test chip. The purpose is to provide.

The present invention was invented in order to achieve the problems and objects in the prior art as described above, and the micro total analysis system of the present invention includes:
A biological substance to be measured contained in a specimen or a treatment liquid obtained by treating the specimen in a flow path;
The reagent stored in the reagent storage unit,
Liquid is sent to the flow path that constitutes the reaction site and merged,
After reacting these, the obtained reaction product or its treated substance
A series of fine flow channels are provided to measure by sending liquid to the flow channel constituting the detection site,
A test chip provided with a pump connection part having a channel opening for communicating with the micropump;
A system main body,
The system main body is
A base body,
Multiple micropumps,
A chip connection portion having a flow path opening for communicating with the inspection chip, and
A micropump unit disposed in the base body,
After mounting the test chip in the base body in a state where the pump connection part of the test chip and the chip connection part of the micro pump unit are in close contact with each other, the reaction between the biological substance and the reagent in the test chip and its A micro total analysis system that automatically performs detection,
In the inspection chip,
A waste liquid storage section is provided for storing a waste liquid generated during the reaction between the biological substance and the reagent and the detection thereof,
It has a water absorption body which holds waste liquid in the waste liquid storage part.

  By configuring in this way, the waste liquid stored in the waste liquid storage unit is securely held by the water absorber, so that the desired liquid can be safely inspected without the waste liquid such as the sample leaking into the system main body. Can do.

Moreover, the micro comprehensive analysis system of the present invention includes:
The water absorbent is
It consists of a water absorbing resin.

  With this configuration, the waste liquid stored in the waste liquid storage unit is securely held by the water-absorbent resin, so that a desired test can be performed safely without causing waste liquid such as a specimen to leak into the system body. be able to.

Moreover, the micro comprehensive analysis system of the present invention includes:
The water absorbing structure is
It consists of sponge.

  By configuring in this way, the waste liquid stored in the waste liquid storage unit is securely held by the sponge, so that a desired test can be safely performed without leakage of the waste liquid such as the specimen into the system main body. it can.

Moreover, the micro comprehensive analysis system of the present invention includes:
The system main body is
After reacting the biological material and the reagent in the fine flow path of the inspection chip,
A detection device that optically performs detection in the flow path that constitutes the detection site;
A pump control device for controlling liquid feeding by the micropump;
And a temperature control device for controlling the temperature at a predetermined part of the inspection chip.

By configuring in this way, it is possible to reliably perform a desired test on the specimen within the system main body.
Moreover, the micro comprehensive analysis system of the present invention includes:
The micropump is
A first flow path in which the flow path resistance changes according to the differential pressure;
A second flow path whose rate of change in flow path resistance with respect to a change in differential pressure is smaller than the first flow path;
A pressurization chamber connected to the first flow path and the second flow path;
An actuator for changing the internal pressure of the pressurizing chamber;
It is characterized by being a micro pump provided with.

With this configuration, the driving liquid can be reliably taken into the inspection chip.
For this reason, the desired test | inspection with respect to a test substance can be reliably performed within a system main body.

Moreover, the micro comprehensive analysis system of the present invention includes:
The micropump is
A positive displacement pump with a pump housing containing a variable volume pump chamber,
The wall forming the pump chamber is composed of a movable deformable wall member such as a flexible diaphragm,
The movement and shape change of the diaphragm changes the volume of the pump chamber,
As a result, the displacement action
The pump chamber has a fluid inlet on the suction side of the pump;
A fluid outlet is provided on the pressure side,
The positive displacement pump is
At least one of the fluid inlet and the fluid outlet is
Comprising a compression component comprising a nozzle in one flow path direction and a diffusion section in the other flow path direction;
The pressure drop across the compression component is
It is characterized in that it is larger in the nozzle direction than in the diffusion direction for one same fluid.

  By configuring in this way, the driving liquid can be reliably taken into the test chip without using a valve at either or both of the fluid inlet and the fluid outlet, and the specimen can be reliably supplied to the specimen within the system body. A desired inspection can be performed.

  According to the present invention, it is possible to provide a micro total analysis system capable of safely performing a desired test without leakage of a waste liquid such as a sample stored in a waste liquid storage part provided at the bottom of a test chip. it can.

Hereinafter, embodiments (examples) of the present invention will be described in more detail with reference to the drawings.
FIG. 1 is a schematic perspective view showing an embodiment of the micro total analysis system of the present invention, and FIG. 2 is a schematic diagram of the inside of the base body showing a state in which a test chip is mounted on the base body of the micro total analysis system of FIG. 3 is a schematic perspective view showing a test chip of the micro total analysis system of FIG. 1, and FIG. 4 is a schematic diagram for explaining a water absorbing body of the micro total analysis system of the present invention.

The micro integrated analysis system 10 shown in FIG. 1 includes a system main body in which a micro pump unit 26, a pump control device 28, a temperature control device 38, a detection device 44 and the like are housed in a base main body 12, and a test chip 50. It is configured.
<Inspection chip>
As schematically shown in FIG. 3, the test chip 50 is connected to the pump connection part 64 provided upstream of the DNA amplification reagent storage part 56a and the chip connection part 66 of the micropump unit 26, The driving liquid 31 is introduced into the inspection chip 50 by the action.

The driving liquid 31 introduced into the test chip 50 sends out the reagent in the DNA amplification reagent storage unit 56a, and further divides the reagent into three parts.
Then, the driving liquid 31 is introduced from upstream of each of the sample DNA storage unit 52a, the positive control storage unit 52b, and the negative control storage unit 52c in which DNA extracted from the sample is stored, and is stored in these storage units. The reagent is mixed with the reagent in the DNA amplification reagent storage unit 56a by pushing out the reagent, and after amplification reaction is performed at a predetermined temperature and time in the amplification reaction unit 57, each is divided into two and sent to the detection unit 39. .

  Further, the DNA amplified by the detection unit 39 is immobilized, and the sample DNA amplification product detection reagent in the sample DNA amplification product detection reagent storage unit 56b and the internal control in the internal control amplification product detection reagent storage unit 56c, respectively. It will be stained with the amplification product detection reagent.

For example, necessary information is extracted by the optical detection device 44 including a light emitting element such as the LED 40 and a light receiving element such as the photodiode 42 to determine the presence or absence of the target DNA.

In addition, downstream of the detection unit 39 is a discharge unit 32 that leads to a waste liquid storage unit for storing the mixed waste liquid after detection.
In addition, each liquid passage is equipped with a water repellent valve for adjusting the timing of air venting and liquid merging, and a mechanism for discarding the leading part where the mixing state is unstable, so that the liquid can flow precisely It is configured.

In addition, such a test | inspection chip | tip 50 is one chip | tip produced by combining suitably one or more members, such as a plastic resin, glass, a silicon | silicone, and ceramics.
Preferably, the fine flow path and the casing of the inspection chip 50 are formed of a plastic resin that is easy to process, inexpensive, and easy to incinerate.

For example, polystyrene resin is excellent in moldability, has a strong tendency to adsorb streptavidin and the like, and can easily form a detection site on a fine channel.
The fine channel is formed to have a width and a depth of, for example, about 10 μm to several hundred μm.

  In addition, in order to optically detect a fluorescent substance or a product of a color reaction, the detection unit 39 that covers at least the detection site of the fine channel on the surface of the test chip 50 is a transparent member, preferably It must be transparent plastic.

The micro integrated analysis system 10 of the present invention can be suitably used particularly for testing genes or nucleic acids.
In this case, the fine flow path of the test chip 50 is configured to be suitable for PCR amplification, but it can be said that the basic flow path configuration is substantially the same for biological materials other than genetic testing.

Usually, the specimen pretreatment unit, reagents, and probes may be changed. In this case, the arrangement and number of liquid feeding elements will change.
A person skilled in the art can easily select the type of analysis by, for example, mounting reagents necessary for an immunoassay method, etc. on the test chip 50, and making modifications including slight changes in flow path elements and specifications. Can be changed.

Biological substances other than genes mentioned here refer to various metabolites, hormones, proteins (including enzymes and antigens), and the like.
In a preferred embodiment of the test chip 50, a sample storage unit into which a sample or an analyte substance (for example, DNA) extracted from the sample is injected in one test chip;
A sample pretreatment unit for preprocessing the sample;
A reagent container for storing reagents used for probe binding reaction, detection reaction (including gene amplification reaction or antigen-antibody reaction);
A positive control accommodating part for accommodating a positive control;
A negative control accommodating portion for accommodating a negative control;
A probe accommodating portion that accommodates a probe (for example, a probe that hybridizes to a gene to be detected amplified by a gene amplification reaction);
A fine flow path communicating with each of these accommodating portions,
A pump connection portion 64 that can be connected to each of the accommodating portions and a separate micropump for feeding the liquid in the flow path is provided.

A micropump is connected to the test chip 50 via a pump connection part 64, and a specimen contained in each accommodation part or a biological substance extracted from the specimen (for example, DNA or other biological substance) and each reagent are accommodated. The reagent contained in the part is sent to the flow path, mixed and reacted at the reaction site of the fine flow path, for example, the site of gene amplification reaction (in the case of protein, antigen-antibody reaction, etc.), and the downstream side The processing solution obtained by processing this reaction solution and the probe stored in the probe storage unit are sent to the detection unit in the channel, mixed in the channel, and combined (or hybridized) with the probe. Biological substances are detected based on the reaction products.

  In addition, the above reaction and detection are performed in the same manner for the positive control housed in the positive control housing section and the negative control housed in the negative control.

The sample DNA storage unit 52a in the test chip 50 performs temporary sample storage and sample supply to the mixing unit.
The sample DNA storage unit 52a is formed with a stopper made of an elastic material such as a rubber-like material or polydimethylsiloxane (PDMS) to prevent leakage, infection and contamination to the outside and to ensure sealing performance. It is desirable to be covered with a resin or a reinforced film.

For example, a specimen in a syringe is injected with a needle that has been pierced with a stopper made of the rubber material or a needle that has passed through a pore with a lid.
In the former case, it is preferable that the needle hole is immediately closed when the needle is pulled out.

Alternatively, another specimen injection mechanism may be installed.
The sample injected into the sample DNA introduction unit 52a is pretreated as necessary, for example, by mixing the sample and the treatment liquid before mixing with the reagent.

Preferred sample pretreatment includes separation or concentration of an analyte (analyte), deproteinization, and the like.
Therefore, the specimen pretreatment unit may include a separation filter, an adsorption resin, beads, and the like.

Necessary reagents are sealed in a predetermined amount in each reagent storage portion of the inspection chip 50 in advance.
Therefore, it is not necessary to fill a necessary amount of the reagent each time it is used, and it can be used immediately.

When analyzing a biological substance in a specimen, reagents necessary for the measurement are generally known.
For example, when analyzing an antigen present in a specimen, a reagent containing an antibody against it, preferably a monoclonal antibody, is used.

The antibody is preferably labeled with biotin and FITC.
Reagents for gene testing may include various reagents used for gene amplification, probes used for detection, and coloring reagent, as well as pretreatment reagents used for the sample pretreatment if necessary.

  In addition, the mixing of the reagent and the reagent and the mixing of the sample and the reagent may be performed at a desired ratio in a single mixing unit, or a plurality of merging units may be provided by dividing either or both of them. Alternatively, they may be mixed so as to obtain a desired mixing ratio.

The mode of such a reactive site is not particularly limited, and various forms and modes are conceivable.
As an example, first from a merging part (flow path branching point) that joins two or more liquids containing a reagent,
A fine flow path in which each liquid is diffused and mixed is provided, and the reaction is performed in a liquid reservoir having a space wider than the fine flow path, provided first from the downstream end of the fine flow path.

As a DNA amplification method, a PCR amplification method that is actively used in various fields can be used.
Various conditions for carrying out the amplification technique have been examined in detail and described in various documents including improvements.

  In the PCR amplification method, it is necessary to manage the temperature by raising and lowering between three temperatures. However, a channel device that enables temperature control suitable for a microchip has already been proposed by the present inventors (Japanese Patent Application Laid-Open No. 2005-318867). 2004-108285).

This device system may be applied to the amplification channel of the chip of the present invention.
As a result, the heat cycle can be switched at high speed, and the micro flow path is a micro reaction cell with a small heat capacity. Therefore, DNA amplification can be performed in a much shorter time than the conventional method that is performed manually.

ICAN (Isothermal chimera) recently developed as an improvement of PCR
The primer initiated nucleic acid amplification) method has a feature that DNA amplification can be performed in a short time at an arbitrary constant temperature of 50 to 65 ° C. (Japanese Patent No. 3343929).

Therefore, the ICAN method is a suitable amplification technique because the inspection chip 50 of the present invention can be simply temperature-controlled.
By hand, the method, which takes one hour, ends with analysis in 10-20 minutes, preferably 15 minutes, in the bioreactor of the present invention.

  In addition, biotin-affinity protein (avidin, streptavidin, extraavidin (R), preferably streptavidin) adsorbed on the detection site on the microchannel is used for biotin labeled on the probe substance or gene amplification reaction. It specifically binds to biotin labeled at the 5 ′ end of the primer.

Thereby, the probe labeled with biotin or the amplified gene is trapped at the detection site.
A method for detecting the separated analyte or the amplified DNA of the target gene is not particularly limited, but as a preferred embodiment, it is basically performed in the following steps.

That is,
(1a) Fine flow channel downstream from the containing portion of a sample or DNA extracted from the sample or cDNA synthesized by reverse transcription reaction from RNA extracted from the sample or sample and a biotin-modified primer at the 5 ′ position To liquid.

  The step of amplifying the gene in the microchannel of the reaction site, the amplification reaction solution containing the gene amplified in the microchannel and the denaturing solution are mixed, and the amplified gene is made into a single strand by denaturation treatment. This is hybridized with the probe DNA fluorescently labeled with FITC (fluorescein isothiocyanate).

Next, the solution is sent to the detection site in the microchannel to which the biotin affinity protein is adsorbed, and the amplified gene is trapped in the detection site in the microchannel. (The amplified gene may be trapped at the detection site and then hybridized with a fluorescently labeled probe DNA.)
(1b) A reagent containing an antibody, preferably a monoclonal antibody, specific to an analyte such as an antigen, metabolite or hormone present in the sample is mixed with the sample.

In that case, the antibody is labeled with biotin and FITC.
Therefore, the product obtained by the antigen-antibody reaction has biotin and FITC.
This is sent to a detection site in a microchannel to which a biotin affinity protein (preferably streptavidin) is adsorbed, and is immobilized on the detection site through the binding of biotin affinity protein and biotin.
(2) A colloidal gold solution whose surface has been modified with an anti-FITC antibody that specifically binds to FITC flows through the fine channel, and this hybridizes to the immobilized FITC of the analyte / antibody reaction product or to the gene. The gold colloid is adsorbed to the FITC-modified probe.
(3) Optically measure the concentration of colloidal gold in the fine channel.
<System itself>
As shown in FIG. 1, the base body 12 has a test chip inlet / outlet port 14 for taking the test chip 50 therein and a display unit for outputting a predetermined test result performed in the base body 12. 18 are provided.

After the inspection chip 50 is placed on the chip transport tray 22, the inspection chip 50 is taken into the base body 12 from the inspection chip entrance 14 and attached.
As shown in FIG. 2, various devices for controlling liquid feeding, reaction, detection and the like in the inspection chip 50 placed on the chip transport tray 22 are provided inside the base body 12. .

Furthermore, a temperature control device 38 including a Peltier element 34 and a heater 36 which are in contact with the inspection chip 50 and controls the temperature of the reagent and the like, particularly the temperature in the reaction part flow path, is provided.
In addition, a detection device 44 is provided that includes an LED 40 that emits light and a photodiode 42 that receives the transmitted light in order to detect a substance to be detected present in the detection portion flow path of the inspection chip 50.

  In the system main body of the micro comprehensive analysis system 10, a micro pump unit 26, a pump control device 28 that controls the micro pump, a temperature control device 38 that controls temperature, a detection device 44, and the like are integrated with the base main body 12.

  When a sample liquid is injected into the sample container 52 of the test chip 50 in which the reagent 58 is sealed in advance and the test chip 50 is attached to the system main body, a mechanical connection for operating the micropump for liquid delivery is made. The

Therefore, when the inspection chip 50 is attached to the system body, the flow path 60 of the inspection chip 50 is in an activated state.
Once the analysis is started, the flow of analytes and reagents, gene amplification based on mixing, reactions such as analyte-probe binding, detection of reactants and optical measurements are automatically performed as a series of sequential steps. The measurement data is stored in a file together with necessary conditions and recorded items, and the measurement of the biological material is automatically performed.

A control system related to each control of liquid feeding, temperature and reaction, a unit responsible for optical detection, data collection and processing constitutes a system main body together with a micropump and an optical device.
This system main body is commonly used for each specimen sample by mounting the test chip 50 on the system main body.

  Reactions such as gene amplification and their detection are performed in software installed in the system body as a program along with micropump and temperature control, optical detection data processing, as preset conditions for liquid delivery sequence, volume, timing, etc. It has been incorporated.

  The detection device 44 that detects the reaction in the fine flow path of the inspection chip 50 irradiates the detection site on the analysis flow path for each inspection item with measurement light from, for example, the LED 40, and the like, and a photodiode 42, a photomultiplier tube, and the like. Transmitted light or reflected light is detected by the optical detection means.

  In the micro integrated analysis system 10 of the present invention, all components are miniaturized and are in a form that is convenient to carry. Therefore, the micro integrated analysis system 10 is excellent in workability and operability without being restricted by the place and time of use.

Since the measurement can be made promptly regardless of the place and time, it can be used for emergency medical care or for personal use in home medical care.
Further, since the micro pump unit 26 used for liquid feeding is incorporated on the system main body side, the inspection chip 50 can be suitably used as a disposable type.

  In the micropump unit 26, for example, the number of parts to be pumped from the upstream side by the driving liquid 31 such as the specimen storage unit 52, the plurality of reagent storage units 56, the positive control storage unit 52b, and the negative control storage unit 52c. A plurality of micropumps are provided correspondingly.

  By mounting the inspection chip 50 on the system main body, the micropump is connected to the inspection chip 50 via the pump connection portion 64 of the inspection chip 50, and functions as a micropump.

In other words, the micropump unit 26 is provided with a plurality of micropumps and a chip connection part 66 having a channel opening for communicating with the inspection chip 50.
On the other hand, the inspection chip 50 is provided with a pump connection part 64 having a flow path opening for communicating with the micropump. The pump connection part 64 of the inspection chip 50 and the chip connection part 66 of the micropump unit 26 are connected to each other. The micropump is communicated with the flow path 60 of the inspection chip 50 by closely contacting.

  Typically, the pump connection portion 64 in the inspection chip 50 includes a channel opening for communicating with the micropump and a contact surface around the channel opening, and the chip connection portion 66 in the micropump unit 26 is connected to the inspection chip 50. It consists of a channel opening for communication and a contact surface around it.

  These are connected by bringing the contact surface on the micropump unit 26 side into contact with the contact surface on the test chip 50 side in a state where the flow path opening on the test chip 50 side matches the flow path opening on the micropump unit 26 side. The

The close contact is performed by pressurizing the inspection chip 50 and the micro pump unit 26, for example.
For example, a sealing member made of a soft resin such as Teflon (registered trademark) is disposed in the chip connecting portion 66 or the pump connecting portion 64, and the sealing surface of the sealing member is used as the inspection chip 50 and the micro pump unit 26. It is good also as a contact surface.

Moreover, it is preferable to use a piezo pump as the micro pump.
In general, the piezo pump has a first flow path in which the flow path resistance changes in accordance with the differential pressure, a second flow path in which the change rate of the flow path resistance with respect to the change in the differential pressure is smaller than the first flow path, A pressurizing chamber connected to the first flow path and the second flow path, and an actuator for changing the internal pressure of the pressurizing chamber are provided. By driving the actuator, the forward direction and the reverse direction are provided. It is a pump that can send liquid.

Details thereof are described in Patent Documents 7 and 8.
According to the above piezo pump, the liquid feeding direction and the liquid feeding speed can be controlled by changing the driving voltage and frequency of the piezo pump.

  As an example, a piezo pump can be manufactured by forming the above pump shape by etching using a silicon substrate obtained by processing a silicon wafer into a predetermined shape by a known photolithography technique and overlaying another substrate thereon. .

The micropump unit 26 in which a large number of piezo pumps are provided on a single silicon substrate can be manufactured by photolithography.
For example, a port is formed on the substrate of the micropump unit 26 and communicates with the pump connection portion 64 of the inspection chip 50 through this port.

  The inspection chip 50 can be connected to the micropump unit 26 by vertically superimposing the substrate with the perforated port and the vicinity of the pump connection portion 64 of the inspection chip 50.

Further, the driving liquid tank 30 is connected to a port opposite to the port connected to the inspection chip 50 via a pump.
When there are a plurality of micropumps, a plurality of ports provided for each of these pumps may be connected to a common drive fluid tank 30.

Furthermore, a known diaphragm pump can be used as the micro pump.
The details of such a diaphragm pump are described in JP-A-8-506874. In general, the diaphragm pump is a displacement pump having a pump housing including a pump chamber for changing the volume, the wall of which is a movable part. Alternatively, a diaphragm is included, and the volume of the pump chamber can be changed by deformation thereof.

The pump chamber has a fluid inlet on the suction side of the pump and a fluid outlet on the opposite pressure side.
In addition, both or one or both of the fluid inlet and the fluid outlet include a compression component that causes a pressure drop in one flow direction to be greater than the opposite flow direction for the same flow.

  Further, by vibrating the diaphragm by the driving means coupled to the diaphragm, the fluid volume in the pump chamber is pulsed, and the fluid can flow through the pump.

The driving means for the pump may be an electromagnetic type in which a plate with a magnet vibrates due to a sine wave of a coil and reciprocates the diaphragm.
With this configuration, the driving liquid can be surely taken into the test chip without using a valve at either or both of the fluid inlet and the fluid outlet, and the specimen can be reliably received in the system body. A desired inspection can be performed.
<Waste liquid storage part>
As shown in FIG. 4, the waste liquid reservoir 46 is a hollow chamber provided at the bottom of the inspection chip 50.

  The waste liquid storage unit 46 is a sealed waste liquid storage that stores all of the excess specimen, the cleaning liquid generated in the specimen separation / concentration process, the waste liquid, and the waste liquid generated as a result of the reaction and measurement of the specimen.

Further, the waste liquid reservoir 46 is provided with a water absorbing body 48 for holding the stored waste liquid, and the stored waste liquid can be reliably held in the water absorbing body 48.
When the sample stored in the sample storage unit 52 is subjected to a sample concentration step as a pre-process in the sample pre-processing unit 53, it is necessary to process a waste liquid generated due to concentration and washing.

It is not bothered to automatically store the waste liquid generated at this time, rather than discharging it to the outside of the inspection chip 50 for processing.
Preferably, the waste liquid storage unit 46 is provided directly below the valve unit 51 of the sample pretreatment unit 53 of the test chip 50 so as to communicate with at least the sample pretreatment unit 53, the reaction site of the flow channel 60, the end of the detection site, and the like. It should be configured.

In addition, the waste liquid storage unit 46 has a sealed structure having a necessary capacity for storing the waste liquid.
That is, it is only necessary to have a cavity in which a through hole communicating with each part that generates waste liquid is formed.

The waste liquid reservoir 46 may be a single hollow chamber or may be in the form of a multi-compartment cavity divided into a plurality of compartments.
The volume, shape, and the like are not particularly limited, but it is necessary to take into consideration the strength of the test chip 50, space limitations associated with the specimen storage unit 52, the reagent storage unit 56, and the like.

For example, the capacity of the waste liquid storage unit 46 is several hundreds μL to 2 mL when the test chip 50 is several cm square and provided at most of the bottom thereof.
In addition, a valve that opens and closes at an appropriate place in the through hole is provided as necessary so that the waste liquid in the waste liquid storage section 46 does not enter other parts of the inspection chip 50 or leak outside. May be.

  In particular, a sample with a low concentration is used by flowing a large amount of sample liquid in the sample pretreatment unit 53, and therefore a relatively large amount of waste liquid including a cleaning solution is generated. A waste liquid storage section 46 having a volume is required.

Thus, in order to make the waste liquid storage part 46 of the test | inspection chip 50 large capacity | capacitance, it is good also considering the whole whole bottom part of the test | inspection chip 50 as a waste liquid storage part which consists of 1 or several hollow structure.
<Water absorber>
In the waste liquid storage part 46, the water absorption body 48 provided therein has a structure that reliably holds the waste liquid to be stored.

In the present embodiment, a water absorbent resin can be used as the water absorbing body 48.
Such a water-absorbing resin is not particularly limited, but is preferably polyvinyl alcohol, polyhydroxyethyl methacrylate, polyethylene glycol, more preferably a polyvinyl alcohol / polyacrylic acid superabsorbent polymer, starch / poly Water-absorbing resins such as acrylic acid graft-based superabsorbent polymers can be used.

Thus, if a water absorbing resin is used as the water absorbing body 48, the waste liquid stored in the waste liquid storage section 46 provided at the bottom of the inspection chip 50 can be reliably held in the water absorbing body 48.
In another embodiment, a sponge can be used for the water absorbing body 48.

  The material of such a sponge is not particularly limited, but a sponge made of polyurethane, non-woven fabric, nylon or the like is preferably used. It is possible to reliably hold the waste liquid.

Further, the water absorbing body 48 is preferably provided in the entire bottom portion of the waste liquid storage section 46, but may be provided in the waste liquid storage section 46 without any gaps.
Furthermore, it may be provided only directly under the flow path of the waste liquid, and the site and the amount of the water absorption body 48 can be appropriately selected according to the amount of the waste liquid discharged from the inspection chip 50.

Further, the installation of the water absorbing body 48 provides the user with the following advantages.
Workers who analyze clinical samples are constantly exposed to the risk of infection from the sample, such as bacteria or viruses.

Not only handling of specimens and pretreatment thereof, but also treatment of washing liquid during measurement, waste liquid after measurement and the like are complicated, and there is still a risk of infection and the possibility of environmental pollution.
The test chip 50 of the present invention has a waste liquid storage section 46 at the bottom of the test chip 50, and has a flow path configuration in which all unnecessary specimens, waste liquid, etc. are finally discarded to the waste liquid storage section 46.

  The waste liquid storage part 46 is completely sealed inside the inspection chip 50 and also holds waste liquid by the water absorbent in the waste liquid storage part 46. The disposable inspection chip 50 is used as it is in the inspection room after use. It is stored in the hazard box provided and incinerated.

  Thus, by providing a water absorbing body such as a water absorbent resin or sponge in the waste liquid storage section 46, after the operator injects the clinical sample into the test chip 50, the operator consistently discards the test after the test. Therefore, there is no possibility of contact with specimens at all, and safety from infection is ensured.

Moreover, disposal can be performed safely without fear of contamination.
In the above embodiment, an example of the micro total analysis system of the present invention has been shown. However, the present invention is not limited to the embodiment, and various modifications can be made without departing from the object of the present invention. It is.

FIG. 1 is a schematic perspective view showing an embodiment of a micro total analysis system of the present invention. FIG. 2 is a schematic diagram for explaining a state in which a test chip is mounted on the base body of the micro total analysis system of the present invention. FIG. 3 is a schematic perspective view for explaining a test chip of the micro total analysis system of the present invention. FIG. 4 is a schematic diagram for explaining a water absorbing body of the micro comprehensive analysis system of the present invention.

Explanation of symbols

DESCRIPTION OF SYMBOLS 10 Micro comprehensive analysis system 12 Base main body 14 Inspection chip entrance / exit 18 Display part 22 Inspection chip conveyance tray 26 Micro pump unit 28 Pump control apparatus 30 Drive liquid tank 31 Drive liquid 32 Discharge part 34 Peltier element 36 Heater 38 Temperature control apparatus 39 Detection part 40 LED
42 Photodiode 44 Optical Detection Device 46 Waste Liquid Storage Unit 48 Water Absorbing Body 50 Test Chip 51 Valve Unit 52 Sample Storage Unit 52a Sample DNA Storage Unit 52b Positive Control Storage Unit 52c Negative Control Storage Unit 53 Sample Pretreatment Unit 56 Reagent Storage Unit 56a DNA Amplification reagent storage unit 56b Specimen amplification product detection reagent storage unit 56c Internal control detection reagent storage unit 57 Amplification reaction unit 58 Reagent 60 Flow path 62 Treatment liquid 64 Pump connection unit 66 Chip connection unit

Claims (6)

A biological substance to be measured contained in a specimen or a treatment liquid obtained by treating the specimen in a flow path;
The reagent stored in the reagent storage unit,
Liquid is sent to the flow path that constitutes the reaction site and merged,
After reacting these, the obtained reaction product or its treated substance
A series of fine flow channels are provided to measure by sending liquid to the flow channel constituting the detection site,
A test chip provided with a pump connection part having a channel opening for communicating with the micropump;
A system main body,
The system main body is
A base body,
Multiple micropumps,
A chip connection portion having a flow path opening for communicating with the inspection chip, and
A micropump unit disposed in the base body,
After mounting the test chip in the base body in a state where the pump connection part of the test chip and the chip connection part of the micro pump unit are in close contact with each other, the reaction between the biological substance and the reagent in the test chip and its A micro total analysis system that automatically performs detection,
In the inspection chip,
A waste liquid storage section is provided for storing a waste liquid generated during the reaction between the biological substance and the reagent and the detection thereof,
A micro total analysis system comprising a water absorbing body for holding waste liquid in the waste liquid storage section. The water absorbent is
The micro total analysis system according to claim 1, comprising a water absorbent resin. The water absorbing structure is
The micro total analysis system according to claim 1, comprising a sponge. The system main body is
After reacting the biological material and the reagent in the fine flow path of the inspection chip,
A detection device that optically performs detection in the flow path that constitutes the detection site;
A pump control device for controlling liquid feeding by the micropump;
The micro integrated analysis system according to claim 1, further comprising a temperature control device that controls a temperature at a predetermined portion of the inspection chip. The micropump is
A first flow path in which the flow path resistance changes according to the differential pressure;
A second flow path whose rate of change in flow path resistance with respect to a change in differential pressure is smaller than the first flow path;
A pressurization chamber connected to the first flow path and the second flow path;
An actuator for changing the internal pressure of the pressurizing chamber;
5. The micro total analysis system according to claim 1, wherein the micro total analysis system is a micro pump equipped with a micro pump. The micropump is
A positive displacement pump with a pump housing containing a variable volume pump chamber,
The wall forming the pump chamber is composed of a movable deformable wall member such as a flexible diaphragm,
The movement and shape change of the diaphragm changes the volume of the pump chamber,
As a result, the displacement action
The pump chamber has a fluid inlet on the suction side of the pump;
A fluid outlet is provided on the pressure side,
The positive displacement pump is
At least one of the fluid inlet and the fluid outlet is
Comprising a compression component comprising a nozzle in one flow path direction and a diffusion section in the other flow path direction;
The pressure drop across the compression component is
5. The micro total analysis system according to claim 1, wherein the same total fluid is larger in the nozzle direction than in the diffusion direction.

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