JP2006275735A - Micro comprehensive analytical system - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To surely secure sufficient tightness in a connection part between a micropump unit and an inspection chip, and to prevent liquid leakage from a flow passage and contamination thereinto, in a micro comprehensive analytical system for performing reaction and detection between a specimen and a reagent in the micro flow passage of an inspection chip, and for carrying out automatically liquid feed, temperature control, detection and the like by attaching the inspection chip to a system main body. <P>SOLUTION: A blower for feeding gas into a connection part between a pump connection part of the inspection chip and a pump connection part of the micropump unit is provided between the pump connection part of the inspection chip and the micro flow passage, and between chip connection part and the micropump unit, and the flow passage is provided in the inspection chip or the micropump unit to introduce the gas delivered from the blower, so as to prevent the liquid leakage from the flow passage and the contamination thereinto. <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 Document 1 to Patent Document 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).
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

In the micro analysis integrated system as described above, a reagent or the like in the fine flow path of the inspection chip is fed by a micro pump.
Specifically, a micro pump unit in which a plurality of pumps using a piezoelectric 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. By closely contacting the chip surface of the micropump unit and the chip surface of the inspection chip, the flow path on the pump side and the flow path on the inspection chip side are communicated with each other.

  However, if the contact surface is wet before connection at the connection between the micropump unit and the inspection chip, liquid such as driving liquid that passes through this connection from the micropump side to the inspection chip at the time of use will be lost. There is a possibility of leakage from between the contact surfaces.

Further, when the liquid tightness is not sufficient at the connection portion between the micro pump unit and the inspection chip, there is a possibility that contamination enters from the outside into the flow path.
In view of the current situation, the present invention can ensure sufficient adhesion at the connection between the micropump unit and the inspection chip, and can prevent liquid leakage from the flow path or entry of contamination. The purpose is to provide an analysis system.

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 channel opening for communicating with the inspection chip, and
A micropump unit disposed in the base body,
The pump connection of the inspection chip;
The chip connection part of the micropump unit;
After mounting the inspection chip in the base body with the liquid tight contact,
A micro total analysis system that automatically performs a reaction between a biological substance and a reagent in the test chip and a detection thereof,
When separating the pump connection part of the inspection chip and the chip connection part of the micro pump unit,
Between the pump connection portion of the inspection chip and the fine flow path,
Between the chip connection part and the micropump unit,
While equipped with a blower device that sends gas to the connection part between the pump connection part of the inspection chip and the chip connection part of the micropump unit,
The inspection chip or the micropump unit is
A flow path for introducing a gas sent out from the blower is provided.

  If comprised in this way, the drive liquid in the flow path near the connection part of a pump connection part and a chip | tip connection part with the gas introduced from the flow path sent out from the air blower and provided in the test | inspection chip or the micropump unit. Can be pushed out of the vicinity of the connection.

For this reason, when the micropump unit and the inspection chip are detached, the liquid can be prevented from flowing out from the joint portion.
Moreover, the micro comprehensive 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 channel opening for communicating with the inspection chip, and
A micropump unit disposed in the base body,
The pump connection of the inspection chip;
The chip connection part of the micropump unit;
After mounting the inspection chip in the base body with the liquid tight contact,
A micro total analysis system that automatically performs a reaction between a biological substance and a reagent in the test chip and a detection thereof,
When separating the pump connection part of the inspection chip and the chip connection part of the micro pump unit,
Between the pump connection portion of the inspection chip and the fine flow path,
Between the chip connection part and the micropump unit,
While equipped with a blower device that sends gas to the connection part between the pump connection part of the inspection chip and the chip connection part of the micropump unit,
The inspection chip and the micropump unit are:
A flow path for introducing a gas sent out from the blower is provided.

  If comprised in this way, the drive liquid in the flow path near the connection part of a pump connection part and a chip | tip connection part with the gas introduced from the flow path sent out from the air blower and provided in the test | inspection chip and the micropump unit. Can be pushed out of the vicinity of the connection.

For this reason, when the micropump unit and the inspection chip are detached, the liquid can be prevented from flowing out from the joint portion.
Moreover, if the flow path for introducing the gas sent out from the blower device is provided on both sides of the inspection chip and the micropump unit, the gas can be introduced from any direction, and the pump connection portion and the chip connection portion The driving liquid in the flow path near the connection portion can be reliably pushed out of the vicinity of the connection portion.

Moreover, the micro comprehensive analysis system of the present invention includes:
The micro total analysis system is
A liquid recovery device is provided for recovering the liquid pushed out of the micropump unit or the inspection chip by the gas fed by the blower.

  With this configuration, the liquid pushed out of the micro pump unit or the inspection chip can be reliably recovered without being scattered in the system main body.

Moreover, the micro comprehensive analysis system of the present invention includes:
The blower is
It is provided in the vicinity of the pump connection part of the inspection chip or the chip connection part of the pump unit.

By comprising in this way, since gas can be directly taken in into the flow path of a test | inspection chip, it can prevent that an apparatus becomes complicated in a system main body.
Moreover, the micro comprehensive analysis system of the present invention includes:
The blower is
It is provided in the vicinity of the pump connection part of the inspection chip and the chip connection part of the pump unit.

By comprising in this way, since gas can be directly taken in into the flow path of a test | inspection chip, it can prevent that an apparatus becomes complicated in a system main body.
Moreover, the micro comprehensive 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 channel opening for communicating with the inspection chip, and
A micropump unit disposed in the base body,
The pump connection of the inspection chip;
The chip connection part of the micropump unit;
After mounting the inspection chip in the base body with the liquid tight contact,
A micro total analysis system that automatically performs a reaction between a biological substance and a reagent in the test chip and a detection thereof,
The pump connection of the test chip;
A hydrophobic layer is provided on the surface of the micropump unit with the chip connection portion.

  In this way, if a hydrophobic layer is provided on the surface of the pump connection portion of the inspection chip and the chip connection portion of the micropump unit, when the inspection chip and the micropump unit are separated from each other, liquid such as driving liquid can be obtained. Since the two-sided tension is used, the liquid can be prevented from scattering other than the connecting portion.

Moreover, the micro comprehensive analysis system of the present invention includes:
The hydrophobic layer is
It is characterized by comprising any of polyethylene, silicone, and Teflon (registered trademark).

  By configuring in this way, when the inspection chip and the micropump unit are separated, the liquid such as the driving liquid is gathered by the double-sided tension, so that it is possible to prevent the liquid from scattering other than the connection portion. .

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;
A temperature control device for controlling the temperature at a predetermined part of the inspection chip; and
It is characterized by providing.

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.

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 showing 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 the embodiment. FIG. 3 is a schematic diagram of the inside of the base body showing a state in which the test chip is mounted on the base body of the micro total analysis system of another embodiment, and FIG. 4 is a diagram showing the main part of the micro total analysis system of FIG. FIG. 5 is a schematic diagram for explaining the test chip of the micro total analysis system of the present invention, and FIG. 6 is an enlarged view of the main part of the test chip of FIG. FIGS. 7 and 7 are schematic diagrams for explaining the movement of the liquid when the blower of the micro total system of the present invention shown in FIG. 2 is used, and FIG. 8 is a diagram of the micro total system of the present invention shown in FIG. Liquid when using a blower Schematic view illustrating the motion, FIG. 9 is a schematic diagram illustrating a micro total system of the present invention, the situation when the is detached the test chip and the micro pump unit of a conventional micro total system.

The micro integrated analysis system 10 shown in FIGS. 1 to 3 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.
<Inspection chip>
The test chip 50 schematically shown in FIG. 5 includes a reagent container 60 (hatched portion in FIG. 5) in which a reagent 58 is sealed in advance, and a flow path 46 extending downstream from the reagent container 60.

  The reagent 58 is supplied from the reagent supply hole 56. When the reagent 58 is supplied into the test chip 50, the reagent 58 is filled up to just before each of the water repellent valve B and the water repellent valve C. It becomes.

Further, a plurality of reagent containers 60 merge at the junction 70 at the downstream side of these reagent containers 60, and a fine channel 46 is provided at the end.
A sample collection unit (not shown) is provided downstream of the microchannel 46, and the sample collected in the sample collection unit and a liquid in which a plurality of reagents 58 passing through the microchannel 46 are mixed. Are further mixed.

In addition, a valve portion such as a water repellent valve is disposed at an appropriate position in the reagent storage unit 60 of the test chip 50, and control of, for example, quantification of the liquid feeding amount and mixing of the liquids is performed.
Further, upstream of the reagent storage unit 60 of the test chip 50, a pump connection unit 64 for connecting to the chip connection unit 66 of the micropump unit 26 shown in FIG. 2 or FIG. An air blower connection part 24 is provided for this purpose.

Such an inspection chip 50 is a single chip produced by appropriately combining one or more members such as plastic resin, glass, silicon, and ceramics.
Preferably, the fine channel 46 and the casing of the inspection chip 50 are formed of a plastic resin that is easy to process, inexpensive, and easy to dispose of by incineration.

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 with 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 portion covering at least the detection portion of the fine flow path on the surface of the inspection chip 50 is a transparent member, preferably a transparent member. It must be plastic.

  Typically, each reagent 58 accommodated in a plurality of reagent accommodating parts 60 is mixed in the downstream reagent accommodating part 60, and further, the mixed reagent in each reagent accommodating part 60 is merged in the merging part 70, and the downstream The liquid is sent to the fine channel 46.

  Although not shown, downstream of the fine channel 46, the specimen and the mixed reagent are merged and mixed from the Y-shaped channel or the like, the reaction is started by temperature rise or the like, and the reaction is performed at the detection site provided in the channel. Detected.

The system of the present invention can be suitably used particularly for testing genes or nucleic acids.
In this case, the fine flow path 46 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 portion into which a sample or an analyte substance (for example, DNA) extracted from the sample is injected in one 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 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 unit 64, and a sample or a biological material extracted from the sample (for example, DNA or other biological material) stored in a sample storage unit (not shown). And the reagent 58 accommodated in the reagent accommodating unit 60 are sent to the merging unit 70 downstream of the reagent accommodating unit 60, and further a reaction site of the fine channel 46, for example, gene amplification reaction (in the case of protein, antigen-antibody reaction, etc.) ), The reaction solution treated with this reaction solution and the probe accommodated in the probe accommodating portion are fed to the detection portion in the downstream flow path, And mixing with the probe (or hybridization), and the biological material is detected based on the reaction product.

  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.

A sample storage unit (not shown) in the test chip 50 communicates with the sample injection unit, and temporarily stores the sample and supplies the sample to the mixing unit.
The specimen injection part that injects the specimen from the upper surface of the specimen storage part (not shown) has a stopper made of an elastic material such as a rubbery material to prevent leakage, infection and contamination to the outside and to ensure sealing performance. Preferably, it is formed or covered with a resin such as polydimethylsiloxane (PDMS) 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 storage unit (not shown) is, for example, a sample and a treatment liquid before being mixed with the reagent 58 in a sample pretreatment unit provided in advance in the reagent storage unit 60 as necessary. Is pretreated by mixing.

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.

The reagent supply hole 56 of the test chip 50 is filled with a predetermined amount of necessary reagents in advance.
Therefore, it is not necessary to fill the necessary amount of the reagent 58 each time it is used, and it is ready for use.

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 58 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.

  By supplying the driving liquid 31 into the test chip 50 by the micro pump of the micro pump unit 26, the reagent 58 is pushed out from each reagent storage unit 60 to the mixing unit 70.

  Furthermore, by pushing out these reagents 58 into the microchannel 46 and merging with the specimen downstream of the microchannel 46, reactions required for analysis such as gene amplification reaction, analyte trap or antigen-antibody reaction are started. Is done.

  The mixing of the reagent 58 and the reagent 58 and the mixing of the sample and the reagent 58 may be performed at a desired ratio in a single mixing section, or a plurality of confluence sections are provided by dividing either or both of them. The final mixing ratio may be 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, a micro flow path 46 in which each liquid is diffused and mixed is provided first from a joining portion (flow path branching point) that joins two or more liquids including the reagent 58, and the downstream side of the micro flow path 46. The reaction is carried out in a liquid reservoir (not shown) comprising a space wider than the fine flow path 46 provided first.

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 the test chip 50 has already been proposed by the present inventors (special feature). No. 2004-108285).

This device system may be applied to the amplification flow path of the inspection chip 50 of the present invention.
As a result, the heat cycle can be switched at high speed, and the micro flow path 46 is a micro reaction cell having a small heat capacity, so that 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.
In the manual operation, this method, which takes one hour, is completed in 10 to 20 minutes, preferably 15 minutes, in the micro integrated analysis system 10 of the present invention.

A detection site for detecting an analyte, for example, an amplified gene, is provided on the downstream side of the reaction site in the fine channel 46 of the test chip 50.
At least the detection part is made of a transparent material, preferably a transparent plastic, in order to enable optical measurement.

  Furthermore, biotin-affinity protein (avidin, streptavidin, extraavidin (R), preferably streptavidin) adsorbed on the detection site on the microchannel 46 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.
The 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. .

  In this embodiment, the pump connection unit 64 connected to the pump connection part 64 of the test chip 50 is provided, and the micropump unit 26 that moves the sample and the processing liquid to a predetermined location and the pump control device 28 that controls the liquid feed of the micropump unit 26 are provided. Yes.

In addition, a temperature control device 38 including a Peltier element 34 and a heater 36 for controlling the temperature of the reagent and the like, particularly the temperature in the reaction part flow path, is provided on the inspection chip 50.
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 is injected into a sample container (not shown) of the test chip 50 in which the reagent 58 has been sealed in advance and the test chip 50 is mounted on the system body, a mechanism for operating a micropump for liquid feeding is used. Connection is made.

Therefore, when the test chip 50 is mounted on the system main body, the reagent storage unit 60 of the test 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 micro flow path 46 of the test chip 50 irradiates the detection site on the analysis flow path for each test item with measurement light from, for example, an LED, and the like, a photodiode, 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.

Moreover, since it can measure rapidly regardless of a place or time, the use in an emergency medical treatment or the personal use in a home medical treatment is also possible.
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, a part to be pumped from the upstream side by the driving liquid 31 such as a specimen storage part (not shown), a plurality of reagent supply holes 56, a positive control storage part, a negative control storage part, etc. A plurality of micropumps are provided corresponding to the number of the micropumps.

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 test chip 50 is provided with a pump connection part 64 having a channel opening for communicating with the micropump as shown in FIG. 4. The pump connection part 64 of the test chip 50 and the micropump unit 26 are provided. The micropump is communicated with the reagent storage unit 60 of the test chip 50 by closely contacting the chip connection unit 66 with the chip connection unit 66.

  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 closely contacting the contact surface on the micropump unit 26 side and the contact surface on the test chip 50 side in a state where the channel opening on the test chip 50 side matches the channel opening on the micropump unit 26 side. Is done.

The close contact is performed by pressurizing the inspection chip 50 and the micro pump unit 26, for example.
A piezo pump is preferably used as the micro pump.

  In general, the piezo pump includes a first flow path in which the flow path resistance changes in accordance with the differential pressure, and 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 that changes the internal pressure of the pressurizing chamber. It is a pump that can feed in the direction.

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 pump driving voltage and frequency.

  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. .

By a photolithography technique, a pump unit in which a large number of piezoelectric pumps are provided on a single silicon substrate can be manufactured.
For example, a port is formed on the substrate of the pump unit and communicated with the pump connection portion 64 of the inspection chip 50 through this port.

The inspection chip 50 can be connected to the pump unit by vertically superimposing the substrate with the perforated port and the vicinity of the pump connection portion 64 of the inspection chip 50.
In addition, the driving liquid tank 30 is connected to the port connected to the inspection chip 50 and the port on the opposite side through the 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.
<Blower device>
In FIG. 2, the inspection chip 50 inserted into the base body 12 while being placed on the chip transport tray 22 is connected at a predetermined position in the base body 12 at the micro pump unit 26 and the pump connection portion 64. Is done.

  Further, the driving liquid 31 such as oil or buffer liquid stored in the driving liquid tank 30 is sent into the test chip 50 by the micro pump driven by the pump control device 28, and is supplied to the reagent storage unit 60 in the test chip 50. The stored reagent 58 or the like is pushed out to the mixing unit 70.

  At this time, if the surface of the chip connection portion 66 of the micro pump unit 26 or the surface of the pump connection portion 64 of the inspection chip 50 is wet with the driving liquid 31, moisture, other liquids, etc., the micro pump unit 26 and the inspection are inspected. Even if the chip 50 is brought into close contact, there is a risk that the driving liquid 31 sent from the micropump leaks out at the connection portion.

  Therefore, in this embodiment, as shown in FIG. 2, the inspection chip 50 is preliminarily formed in a stage before the chip connection portion 66 of the connected micro pump unit 26 and the pump connection portion 64 of the inspection chip 50 are separated. Gas is sent from the blower 32 into the provided flow path 20 and the driving liquid 31 is removed in advance to prevent the liquid from leaking out from these connecting portions.

As shown in FIG. 2, such a blower 32 is configured to be connected to a blower connection unit 48 provided in advance on the inspection chip 50 in the system main body.
Further, the micropump unit 26 connected to the pump connection portion 64 of the inspection chip 50 may be provided with a liquid recovery device connection portion 49 as shown in FIG. 49 is connected to the liquid recovery device 16.

  As shown in FIG. 4, such a liquid recovery device 16 takes gas into the flow path 20 from the blower device 32 in a state where the micropump unit 26 and the inspection chip 50 are connected, and further drives the driving liquid 31. Is pushed up to the liquid recovery device connection portion 49 of the upper micropump unit 26, and the drive liquid 31 is recovered by the liquid recovery device 16 connected to the liquid recovery device connection portion 49.

  Note that when the gas is taken into the inspection chip 50 by the blower 32, the flow path 59 and the flow path 71 have different diameters as shown in FIG. It is configured so that it can be surely captured.

  Similarly, on the micro pump unit 26 side, the flow diameter of the drive liquid pushed up by the gas sent from the blower 32 is changed by making the flow path on the drive liquid tank 30 side different from the flow path on the liquid recovery apparatus side. 31 is configured to be reliably recovered by the liquid recovery device 16.

  As shown in FIG. 7A, the flow of recovery of the driving liquid 31 by the blower 32 is such that the chip connection portion 66 of the micropump unit 26 and the pump connection portion 64 of the inspection chip 50 are connected. In this state, the blower 32 is connected to the blower connection part 48 of the inspection chip 50.

  In this state, when the gas is sent into the flow path 20 from the blower 32, the driving liquid 31 filled in the flow path 59 is pushed upward by the gas 68 as shown in FIG.

When the gas is further fed into the flow path, the driving liquid 31 in the flow path 59 does not move from the vicinity of the chip connection portion 66 and the pump connection portion 64 as shown in FIG. 7C. .
In this way, if the micro pump unit 26 and the inspection chip 50 are detached in the state where the driving liquid 31 is not present in the vicinity of the chip connection part 66 and the pump connection part 64, liquid can be reliably prevented from leaking at the connection part. can do.

  Further, when the liquid recovery device connecting portion 49 is provided in the micro pump unit 26, as shown in FIG. 8A, the chip connecting portion 66 of the micro pump unit 26 and the pump connecting portion 64 of the inspection chip 50 are provided. Are connected to the blower connection portion 48 of the test chip 50.

Further, the liquid recovery apparatus 16 is connected to the liquid recovery apparatus connection portion 49 of the micropump unit 26.
In this state, when the gas is sent from the blower 32 into the flow path 20, the driving liquid 31 filled in the reagent container 60 is pushed upward by the gas 68 as shown in FIG. .

  When the gas is further fed into the flow path, the driving liquid 31 in the reagent container 60 is connected to the liquid recovery device connection part 49 in the vicinity of the chip connection part 66 and the pump connection part 64 as shown in FIG. Thus, the liquid is recovered in the liquid recovery device 16.

  Note that the type, amount, and temperature of the gas sent out by the blower 32 may be appropriately set in accordance with the driving liquid 31, the specimen or processing liquid contained in the test chip 50, the configuration of the system body, and the like.

The number of blower devices 32 can be set as appropriate according to the number of blower connection portions 24 provided on the inspection chip.
As such an installation example, one air blower 32 is installed, and the air blower 32 and all the air blower connection parts 24 are connected to each other so that gas is sent from one air blower, or a plurality of air blowers An installation example is provided in which the device 32 is installed, and each of the air blowers 32 is shared and connected to the air blower connection portion 24 of the inspection chip 50 so as to send in gas.

  Further, in the present embodiment, the blower 32 is connected to the inspection chip 50 side, and gas is allowed to flow in from the inspection chip 50 side, thereby eliminating the driving liquid 31 in the flow path near the chip connection portion 66 and the pump connection portion 64. The case where the air blower 32 is connected to the inspection chip 50 side, and the case where the liquid recovery device 16 on the micro pump unit 26 side collects the driving liquid 31 is described. It is not limited to this.

  For example, the blower 32 is connected to the micro pump unit 26 side, and the driving liquid 31 in the flow path near the chip connection portion 66 and the pump connection portion 64 is eliminated by allowing gas to flow from the micro pump unit 26 side. Alternatively, the air blower 32 may be connected to the micro pump unit 26 side, and the liquid recovery device 16 may be provided on the test chip 50 side to recover the liquid.

Further, the blower device may be configured to be switchable so that it can be used on both the micropump unit 26 side and the inspection chip 50 side, and can be changed without departing from the content of the present invention.
<Hydrophobic layer>
A hydrophobic layer 52 can be provided on the surface of the chip connection part 66 or the pump connection part 64.

Conventionally, as shown in FIG. 9 (a-1), after the micropump unit 26 and the test chip 50 are brought into close contact with each other and the driving liquid 31 is caused to flow into the reagent storage unit 60, as shown in FIG. 9 (a-2). As described above, the micropump unit 26 and the inspection chip 50 are separated.

  At this time, the driving liquid 31 in the vicinity of the surfaces of the pump connection portion 64 and the chip connection portion 66 is connected between the respective surfaces during the initial separation, and further, between the pump connection portion 64 and the chip connection portion 66. The liquid was drained by increasing the distance.

When the liquid is drained, the liquid may be scattered in the system main body or on the inspection chip 50.
In the embodiment of the present invention, as shown in FIG. 9 (b-1), the hydrophobic layer 52 is provided on the surfaces of the pump connection part 64 and the chip connection part 66, and in this state, the micro pump unit 26 and the inspection chip are provided. 50.

  Further, when the micropump unit 26 and the inspection chip 50 are separated, the surface tension of the driving liquid 31 is generated by the hydrophobizing layer 52 provided on the surfaces of the pump connection part 64 and the chip connection part 66, and the liquid draining is performed. Can be done smoothly.

  Such a hydrophobic layer 52 may be a sealing member made of a soft resin such as polyethylene, silicone, or Teflon (registered trademark), and the sealing surface of the sealing member is connected to the inspection chip 50, the micro pump unit 26, and the like. It is good to use the contact surface.

  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 view of the inside of the base body showing a state in which the inspection chip is mounted on the base body of the micro total analysis system of the embodiment. FIG. 3 is another schematic view of the inside of the base body showing a state in which the test chip is mounted on the base body of the micro total analysis system of another embodiment. FIG. 4 is an enlarged view of the main part showing the main part of the micro comprehensive analysis system of FIG. 3 in an enlarged manner. FIG. 5 is a schematic diagram for explaining a test chip of the micro comprehensive analysis system of the present invention. FIG. 6 is an enlarged view of a main part of the main part of the inspection chip shown in FIG. FIG. 7 is a schematic diagram for explaining the movement of the liquid when using the blower of the micro total system of the present invention shown in FIG. FIG. 8 is a schematic diagram for explaining the movement of the liquid when using the blower of the micro total system of the present invention shown in FIG. FIG. 9 is a schematic diagram for explaining the situation when the micro integrated system of the present invention, the inspection chip of the conventional micro integrated system, and the micro pump unit are detached.

Explanation of symbols

DESCRIPTION OF SYMBOLS 10 Micro integrated analysis system 12 Base main body 14 Test | inspection chip entrance / exit 16 Liquid collection | recovery apparatus 18 Display part 20 Flow path 22 Test | inspection chip conveyance tray 24 Blower connection part 26 Micro pump unit 28 Pump control apparatus 30 Drive liquid tank 31 Drive liquid 32 Blower 34 Peltier element 36 Heater 38 Temperature controller 40 LED
42 Photodiode 44 Detection Device 46 Fine Flow Channel 48 Blower Device Connection Portion 49 Liquid Recovery Device Connection Portion 50 Inspection Chip 52 Hydrophobized Layer 56 Reagent Supply Hole 58 Reagent 59 Flow Channel 60 Reagent Storage Portion 62 Treatment Solution 64 Pump Connection Portion 66 Chip Connection Portion 68 Gas 70 Merge portion 71 Flow path B Water repellent valve portion C Water repellent valve portion

Claims (9)

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 channel opening for communicating with the inspection chip, and
A micropump unit disposed in the base body,
The pump connection of the inspection chip;
The chip connection part of the micropump unit;
After mounting the inspection chip in the base body with the liquid tight contact,
A micro total analysis system that automatically performs a reaction between a biological substance and a reagent in the test chip and a detection thereof,
When separating the pump connection part of the inspection chip and the chip connection part of the micro pump unit,
Between the pump connection portion of the inspection chip and the fine flow path,
Between the chip connection part and the micropump unit,
While equipped with a blower device that sends gas to the connection part between the pump connection part of the inspection chip and the chip connection part of the micropump unit,
The inspection chip or the micropump unit is
A micro total analysis system comprising a flow path for introducing a gas sent from the blower. 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 channel opening for communicating with the inspection chip, and
A micropump unit disposed in the base body,
The pump connection of the inspection chip;
The chip connection part of the micropump unit;
After mounting the inspection chip in the base body with the liquid tight contact,
A micro total analysis system that automatically performs a reaction between a biological substance and a reagent in the test chip and a detection thereof,
When separating the pump connection part of the inspection chip and the chip connection part of the micro pump unit,
Between the pump connection portion of the inspection chip and the fine flow path,
Between the chip connection part and the micropump unit,
While equipped with a blower device that sends gas to the connection part between the pump connection part of the inspection chip and the chip connection part of the micropump unit,
The inspection chip and the micropump unit are:
A micro total analysis system comprising a flow path for introducing a gas sent from the blower. The micro total analysis system is
3. The micro total analysis system according to claim 2, further comprising a liquid recovery device that recovers the liquid pushed out of the micro pump unit or the inspection chip by the gas sent by the blower. The blower is
4. The micro total analysis system according to claim 1, wherein the micro total analysis system is provided in the vicinity of a pump connection part of the inspection chip or a chip connection part of the pump unit. 5. The blower is
The micro total analysis system according to any one of claims 1 to 3, wherein the micro total analysis system is provided in the vicinity of a pump connection portion of the inspection chip and a chip connection portion of the pump unit. 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 channel opening for communicating with the inspection chip, and
A micropump unit disposed in the base body,
The pump connection of the inspection chip;
The chip connection part of the micropump unit;
After mounting the inspection chip in the base body with the liquid tight contact,
A micro total analysis system that automatically performs a reaction between a biological substance and a reagent in the test chip and a detection thereof,
The pump connection of the test chip;
A micro total analysis system, wherein a hydrophobic layer is provided on a surface of the micro pump unit with the chip connection portion. The hydrophobic layer is
The micro total analysis system according to claim 6, which is made of any one of polyethylene, silicone, and Teflon (registered trademark). 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;
A temperature control device for controlling the temperature at a predetermined part of the inspection chip; and
The micro total analysis system according to any one of claims 1 to 7, further comprising: 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;
The micro comprehensive analysis system according to claim 1, wherein the micro total analysis system is a micro pump equipped with a micro pump.

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