WO2005108571A1 - 検査用マイクロリアクタおよび遺伝子検査装置ならびに遺伝子検査方法 - Google Patents
検査用マイクロリアクタおよび遺伝子検査装置ならびに遺伝子検査方法 Download PDFInfo
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- B01F35/717—Feed mechanisms characterised by the means for feeding the components to the mixer
- B01F35/7176—Feed mechanisms characterised by the means for feeding the components to the mixer using pumps
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- B01F35/00—Accessories for mixers; Auxiliary operations or auxiliary devices; Parts or details of general application
- B01F35/80—Forming a predetermined ratio of the substances to be mixed
- B01F35/83—Forming a predetermined ratio of the substances to be mixed by controlling the ratio of two or more flows, e.g. using flow sensing or flow controlling devices
- B01F35/831—Forming a predetermined ratio of the substances to be mixed by controlling the ratio of two or more flows, e.g. using flow sensing or flow controlling devices using one or more pump or other dispensing mechanisms for feeding the flows in predetermined proportion, e.g. one of the pumps being driven by one of the flows
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- B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
- B01L3/5027—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip
- B01L3/502723—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip characterised by venting arrangements
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- B01L3/5027—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip
- B01L3/502738—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip characterised by integrated valves
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Definitions
- the present invention relates to a microreactor, particularly to a genetic test device including a bioreactor that can be suitably applied to a genetic test.
- Patent Document 1 JP 2001-322099 A
- Patent Document 2 Japanese Patent Application Laid-Open No. 2004-108285
- Non-Patent Document 1 "DNA Chip Technology and Its Applications”, “Protein Nucleic Acid Enzyme”, Vol. 43, No. 13 (1998) Fumio Kimizuka, Ikunoyuki Kato, Kyoritsu Shuppan Co., Ltd.
- the present invention provides a microreactor which is a disposable type, low cost, has a simple configuration and a highly accurate liquid sending system, and enables highly accurate detection, particularly a microreactor for genetic testing. The purpose is to do.
- Another object of the present invention is to provide a bio-microreactor having a cross-contamination and a carry-over contaminant, which is unlikely to cause problems.
- the gene testing apparatus of the present invention has been made in view of the above-described circumstances, and has a method of appropriately changing primers and bioprobes in order to ensure versatility and high sensitivity.
- a reagent mixing unit that mixes a plurality of reagents delivered from outlets of the plurality of reagent storage units to generate a mixed reagent
- reaction section for mixing and reacting the mixed reagent delivered from the reagent mixing section and the sample delivered from the sample receiving section,
- the plurality of reagent storage units, the reagent mixing unit, the sample receiving unit, and the reaction unit are incorporated in the chip, and are communicated with each other by a flow path.
- the reagent mixing section has a delivery prevention mechanism for preventing delivery of the initial mixed reagent to the reaction section.
- the reagent mixing section forms a mixing channel, and a delivery channel for delivering the mixed reagent to the reaction section is branched at an intermediate portion of the mixing channel, and the initial mixed reagent is It is accommodated between the middle part and the downstream end of the mixing channel and is prevented from being sent from the sending channel to the reaction section.
- the reagent mixing section is connected to the connection section between the mixing channel and the sending channel when the pressure in the mixing channel becomes equal to or higher than a predetermined pressure. It has a liquid sending control unit that sends out the mixed reagent.
- the microreactor for sample inspection is
- a reagent mixing unit that mixes a plurality of reagents delivered from outlets of the plurality of reagent storage units to generate a mixed reagent
- reaction section for mixing and reacting the mixed reagent delivered from the reagent mixing section and the sample delivered from the sample receiving section
- the plurality of reagent storage units, the reagent mixing unit, the sample receiving unit, and the reaction unit are incorporated in the chip, and are communicated with each other by a flow path.
- Each of the reagent storage units has an inlet for injecting the driving liquid into the storage chamber and an outlet for pushing out the reagent in the storage chamber by the injected reagent, and the injection port is a pump connectable to an external pump. After being connected to the connecting portion, the driving liquid is injected into the receiving chamber from the inlet by an external pump, and an air vent channel having an open end is provided at a connecting portion between the inlet and the pump connecting portion.
- FIG. 1 is a schematic view of a microreactor for genetic testing according to one embodiment of the present invention.
- FIG. 2 is a schematic diagram of a genetic test device including the microreactor of FIG. 1 and a device main body.
- Fig. 3 is a diagram showing a state in which a sealant is filled between the reagent accommodating section and a flow path communicating therewith.
- FIG. 4 shows a piezo pump
- (a) is a cross-sectional view showing an example of the pump
- (b) is a top view thereof.
- (C) is a sectional view showing another example of the piezo pump.
- FIG. 5 is a graph showing a relationship between a drive voltage waveform applied to a piezoelectric element of a pump and a displacement of a liquid.
- FIG. 6 (a) is a diagram showing a configuration of a pump unit for sending a driving liquid
- FIG. 6 (b) is a diagram showing a configuration of a pump unit for sending a reagent.
- FIG. 7 is a view showing a flow path for air release.
- FIGS. 9 (a) and 9 (b) are cross-sectional views along the flow channel axis direction of a liquid sending control unit.
- FIG. 10 (a) and (b) are cross-sectional views showing an example of a check valve provided in a flow path. [FIG.
- FIG. 11 is a cross-sectional view showing an example of an active valve provided in a flow path, where (a) shows an open valve state,
- FIG. 12 is a diagram showing a configuration of a reagent quantification unit.
- Fig. 13 is a view showing a flow path configuration in which the head portion is truncated so that the mixture ratio is stabilized and the mixture is supplied to the next step.
- FIG. 14 is a diagram showing a configuration of a reagent mixing section of the microreactor according to one embodiment of the present invention.
- FIG. 15 is a diagram showing a configuration of a portion that communicates with the flow channel of FIG. 14 and performs amplification reaction and detection of a sample and a reagent.
- FIG. 16 is a diagram showing a configuration of a part that performs amplification reaction and detection between a positive control and a reagent, which also communicates with the flow channel shown in FIG. 14.
- FIG. 17 is a diagram showing a configuration of a part for performing amplification reaction and detection of a negative control and a reagent, which also communicates with the flow channel shown in FIG. 14.
- FIG. 18 is a cross-sectional view showing an example of an active valve provided in a flow path, where (a) shows an open state and (b) shows a closed state.
- FIG. 19 is a cross-sectional view showing an example of an active valve provided in a flow channel, where (a) shows a valve open state and (b) shows a valve closed state.
- the microreactor for genetic testing of the present invention comprises:
- a sample container into which a sample or DNA extracted from the sample is injected,
- a reagent storage section in which a reagent used for the gene amplification reaction is stored
- a positive control housing section in which the positive control is housed is housed
- a negative control housing section in which the negative control is housed is housed
- a probe DNA storage unit in which a probe DNA to be hybridized to the gene to be detected amplified by the gene amplification reaction is stored;
- a pump connection unit that can be connected to a separate micropump that sends the liquid in each of the storage units and the flow path;
- a micropump is connected to the chip via a pump connector, and the sample or the sample DNA extracted from the sample container and the reagent stored in the reagent container are sent to the channel. After mixing and amplifying in the flow channel, the processing solution obtained by treating this reaction solution and the probe DNA stored in the probe DNA storage section are sent to the downstream flow channel. In the reaction mixture, hybridization is performed, and based on the reaction product, an amplification reaction is detected.
- the microreactor for genetic testing comprises:
- a reverse transcriptase storage unit in which a sample or RNA extracted from the sample is injected into the sample storage unit, and a reverse transcriptase for synthesizing cDNA by reverse transcription reaction from the RNA contained therein is provided,
- the sample contained in the sample storage unit or the RNA extracted from the sample and the reverse transcriptase stored in the reverse transcriptase storage unit are sent to the flow channel and mixed in the flow channel to synthesize cDNA.
- the amplification reaction and the detection thereof may be performed.
- microreactor for genetic testing comprises:
- the pump of the microphone port pump wherein the passage of the liquid is blocked until the liquid sending pressure in the forward direction reaches a preset pressure, and the liquid is allowed to pass by applying a liquid sending pressure higher than the preset pressure.
- a liquid sending control unit capable of controlling the passage of liquid by pressure
- a backflow prevention unit for preventing backflow of the liquid in the flow path is provided,
- the micro pump, the liquid sending control unit, and the backflow prevention unit control the liquid sending, the fixed amount, and the mixing of the liquids in the flow path.
- the liquid sending control section is formed of a fine channel having a cross-sectional area smaller than the cross-sectional area of these adjacent flow channels formed between these flow channels so as to connect adjacent flow channels on both sides in series.
- the microreactor for genetic testing described above comprises:
- the backflow prevention portion may be a check valve in which the valve body closes the flow path opening by the backflow pressure, or an active valve that closes the opening by pressing the valve body against the flow path opening by the valve body deformation means. It is characterized by.
- a reagent-filled flow path that is configured with a flow path between the backflow prevention unit and the liquid-feed control unit, and that can be filled with a predetermined amount of reagent;
- a branch flow path is provided, which is branched from the reagent filling flow path and communicates with a pump connection portion connected to a micropump that feeds a driving liquid.
- the reagent does not pass from the backflow prevention unit side to the liquid supply control unit.
- the reagent is The micropump is used to supply the driving liquid in a direction facing the reagent filling flow path at a liquid sending pressure that allows the reagent to pass therethrough, thereby filling the reagent filling flow path.
- There is provided a reagent quantitative section configured to push out the reagent from the liquid sending control section first, thereby quantitatively sending the reagent.
- a mixing channel connected to the plurality of channels and mixing each reagent from the channels;
- a second liquid supply control unit is provided at a position near the branch point of the branch flow path with the mixing flow path, and the liquid supply pressure at which the reagent mixture can pass is smaller than the first liquid supply control unit.
- the reagent mixture is sent until the tip of the reagent mixture sent into the mixing channel reaches the first solution sending control unit, and then the reagent mixture solution passes through the first solution sending control unit.
- the configuration is such that the reagent mixture is passed from the second liquid supply controller to the branch flow path at the liquid supply pressure, and the reagent mixture is supplied to the next step.
- the microreactor for genetic testing described above comprises:
- the cross-sectional area force of the narrow flow path in the first liquid flow control unit is smaller than the cross-sectional area of the narrow flow path in the second liquid flow control unit.
- the microreactor for genetic testing is characterized in that a flow path between the pump connection portion and the storage portion in which the content liquid sent by the micropump connected to the pump connection portion is stored. It is characterized in that a flow path for air bleeding, which branches off from the road and has an open end, is provided.
- Reagents used for gene amplification reaction, positive control and negative control It is preferable that the file is accommodated in the accommodation section.
- the liquid content in the storage section before use is placed between each storage section for storing the reagent used for the gene amplification reaction, the positive control and the negative control, and the channel communicating therewith. It is characterized by being filled with a sealant for preventing leakage into the flow path.
- the sealant preferably has an oil-and-fat power having a solubility in water of 1% or less.
- the sealant has a solubility in water of 1% or less and a melting point of 8 ° C to room temperature (25 ° C).
- sealing agent an aqueous solution of gelatin is preferable.
- the microreactor for genetic testing described above comprises:
- the reagent used for the gene amplification reaction comprises a chimeric primer that specifically hybridizes to the gene to be detected, a DNA polymerase having strand displacement activity, and an endnuclease.
- the sample or the sample extracted from the sample, or the cDNA synthesized by reverse transcription reaction from the sample or the sample extracted from the sample and the RNA, and the biotin-modified primer are sent to the flow channel from these storage sections and Performing a gene amplification reaction in the road;
- the genetic testing device of the present invention includes the microreactor and a micropump connected to a pump connection of the microreactor.
- the genetic test apparatus is
- the micropump a first flow path in which the flow path resistance changes according to the differential pressure
- the genetic test apparatus is
- a pump connection portion is provided on the upstream side of each reagent storage portion in which a reagent is stored, and a micropump is connected to these pump connection portions, and each micropump power is supplied by a driving liquid, whereby the power of the reagent storage portion is increased.
- the genetic test apparatus is
- the reagent is mixed at a desired ratio by controlling the operation of the actuator by a driving signal of a driving device of the micropump.
- the above-described genetic test apparatus preferably includes a detection apparatus for detecting an amplification reaction based on a reaction product of hybridization between the amplified gene and probe DNA.
- the above-described genetic testing device preferably includes a temperature control device for controlling a reaction temperature of each reaction in the flow channel of the microreactor.
- the above-described genetic testing device comprises a device main body in which the micropump, the detection device, and the temperature control device are integrated, and a microreactor that can be mounted on the device main body. It is characterized in that the gene amplification reaction and the detection of the amplification reaction are performed automatically.
- the microreactor of the present invention has a configuration suitable for mass production, and has a multi-purpose power. Due to its utility, it can be manufactured at low cost. In addition, since the flow path system including the pump and the valve has a simple configuration, the dead volume through which air bubbles enter is small, and the liquid sending precision is high. This microreactor enables detection with high detection sensitivity because it incorporates a DNA amplification step for detection.
- the genetic test apparatus of the present invention has a system configuration in which the reagents for each sample 'components equipped with a liquid-sending system element and the control' detection components are separated from each other. And serious problems such as cross-contamination and carryover-one contamination. Since it is easy to wash and remove nonspecific binding substances other than the binding (or interaction) between the sample DNA and the primers and probes, a low knock ground and a microreactor chip can be provided.
- the present invention provides gene expression analysis, gene function analysis, single gene polymorphism analysis (SNP), drug screening, safety and toxicity testing of drugs, pesticides, and various danigaku substances, clinical clinical diagnosis, and food Applicable in fields such as inspection, forensic medicine, chemistry, brewing, agriculture and forestry, fisheries, livestock and agricultural production.
- SNP single gene polymorphism analysis
- microreactor of the present invention a microreactor of the present invention
- a genetic test apparatus including the microreactor, each control device, and a detection device
- a gene test method including a gene amplification step and a detection step using the present apparatus
- FIG. 1 is a schematic diagram of a microreactor for genetic testing according to one embodiment of the present invention
- FIG. 2 is a schematic diagram of a genetic testing device including the microreactor and a device main body according to one embodiment of the present invention.
- the microreactor shown in Fig. 1 is composed of a single chip made of resin, glass, silicon, ceramics, or the like.
- the chip includes a sample storage section, reagent storage section, probe DNA storage section, control storage section, flow path, pump connection section, liquid transfer control section, backflow prevention section, Each part of the drug metering section and the mixing section is installed at a functionally appropriate position by micromachining technology. If necessary, a reverse transcriptase section may be provided.
- the sample storage unit communicates with the sample injection unit to temporarily store the sample and supply the sample to the mixing unit. In some cases, it may have the function of separating blood cells.
- the mixing of the reagent with the reagent and the mixing of the sample with the reagent may be performed in a single mixing section at a desired ratio, or one or both may be divided to provide a plurality of junctions. It may be mixed so as to have a desired mixing ratio.
- the gene amplification reaction and the processing necessary for its detection are automatically performed in the chip, and the gene can be simultaneously and quickly analyzed for a large number of items. It is configured to allow inspection.
- necessary reagents are pre-packaged in a predetermined amount, and DNA or RNA of a sample and a predetermined amplification reaction, and detection of an amplification product are detected.
- the microreactor is used for each sample as a unit for performing the above.
- a control system, optical detection, data collection, and processing related to each control of liquid feeding, temperature, and reaction constitute a main body of the genetic testing device of the present invention together with the micropump and the optical device.
- the main body of the apparatus is commonly used for a specimen sample by mounting the chip on the main body. Therefore, even if there are many samples, it can be processed efficiently and quickly.
- the present invention only the above-mentioned detachable chip needs to be replaced. If it is necessary to change the control of each device element, the control program stored in the main body of the device must be appropriately modified.
- the genetic test apparatus of the present invention is small in size and convenient to carry, and therefore has good workability and operability regardless of the place and time of use.
- the sample to be measured according to the present invention is a gene, DNA or RNA as a nucleic acid that becomes a type II amplification reaction in the case of a genetic test. It may be prepared or isolated from a sample that may contain such a nucleic acid.
- the method for preparing a gene, DNA or RNA with such a sample power is not particularly limited, and conventional techniques can be used. Recently, techniques for preparing genes, DNAs or RNAs from biological samples for DNA amplification have been developed and can be used in the form of kits and the like.
- the sample itself is not particularly limited, but most samples derived from living organisms such as whole blood, serum, buffy coat, urine, feces, saliva, sputum, etc .; cell cultures; viruses, bacteria, molds, and yeasts Examples include nucleic acid-containing samples of plants, animals, and the like; samples that may contain or contain microorganisms and the like, and any other samples that may contain nucleic acids.
- DNA can be separated and purified from a sample according to a conventional method by phenol-form extraction with chloroform and ethanol precipitation. It is generally known to use high concentrations of chaotropic reagents, such as guanidine hydrochloride or isothiocyanate, which are near saturating concentrations, to release nucleic acids. Instead of applying the above phenol-chloroform extraction method, etc., directly treat the specimen with a protease-containing protease solution (Takashi Saito, ⁇ PCR Experiment Manual '', HBJ Press, 1991) , P309) is a simple and fast method.
- a protease-containing protease solution Takashi Saito, ⁇ PCR Experiment Manual '', HBJ Press, 1991
- the obtained genomic DNA or gene may be fragmented using an appropriate restriction enzyme, for example, BamHI, BgLII, Dral, EcoRI, EcoRV, HindIII, PvuII, etc. according to a conventional method. In this way, an aggregate of the sample DNA and its fragments can be prepared.
- an appropriate restriction enzyme for example, BamHI, BgLII, Dral, EcoRI, EcoRV, HindIII, PvuII, etc.
- RNA molecules of a retrovirus functioning as a gene are targeted.
- RNA molecules of a retrovirus functioning as a gene are targeted.
- RNA and other RNA molecules such as mRNA and rRNA, which are direct communication carriers for the expressed gene, are targeted.
- These RNAs may be converted to cDNA using an appropriate reverse transcriptase and the force analyzed.
- the method for preparing mRNA should be based on known techniques. And reverse transcriptase is readily available.
- the microreactor of the present invention requires an extremely small amount of sample as compared with a manual operation performed using a conventional apparatus. For example, in the case of a gene, the DNA is 0.001 to 100ng. For this reason, the microreactor of the present invention, even when only a small amount of sample is obtained, is less restricted from the aspect of the specimen and inevitably requires a smaller amount of reagents, thereby reducing the test cost.
- the sample is introduced from the injection part of the “sample storage part”.
- the amplification method is not limited.
- a DNA amplification technique a PCR amplification method that is widely used in various fields can be used.
- Various conditions for implementing the amplification technology have been studied in detail, and are described in various documents, including improvements.
- PCR amplification it is necessary to control the temperature by raising and lowering the temperature between three temperatures.
- the present inventors have already proposed a flow path device capable of controlling the temperature suitable for a microchip (see Japanese Patent Application Laid-Open 2004—108285). This device system may be applied to the channel for amplification of the chip of the present invention.
- the thermal cycle is switched at high speed, and the microchannel is a micro reaction cell with a small heat capacity, so DNA amplification is performed in a much shorter time than in the conventional method using manual micro tubes and micro vials. be able to.
- the Isotnermal chimera primer initiated nucleic acid amplification method is characterized in that DNA amplification can be performed in a short time at an arbitrary constant temperature of 50 to 5 ° (Patent No. 3433929). Therefore, the ICAN method is a suitable amplification technique in the microreactor of the present invention because simple temperature control is sufficient. By hand, the method, which takes one hour, ends up to be analyzed in 10-20 minutes, preferably 15 minutes, in the bioreactor of the invention.
- the microreactor of the present invention in which the DNA amplification reaction may be performed by another PCR method, has the flexibility to cope with any of them by changing the design of the flow path.
- the details of the technique for using any of the DNA amplification reactions are disclosed, and can be easily derived by those skilled in the art.
- PCR primers are two types of oligonucleotides that are amplified and complementary to both ends of a DNA strand at a specific site.
- Primers for the ICAN method are chimeric primers of DNA and RNA, and their preparation has already been technically established (Patent No. 3433929). It is important to use the most appropriate primer design and selection to determine the success or failure of the amplification reaction and the efficiency.
- the DNA of the amplification product can be immobilized on the substrate via binding to streptavidin on the substrate, and can be used for quantification of the amplification product.
- primer labeling substances include digoxigenin and various fluorescent dyes.
- Reagents such as enzymes used in amplification reactions can be easily obtained for both PCR and ICAN.
- the reagents in the PCR method include at least the power of 2'-deoxynucleoside 5'-triphosphate, Taq DNA polymerase, Vent DNA polymerase or Pfo DNA polymerase.
- the reagents in the ICAN method include at least 2'-deoxynucleoside 5'-triphosphate, a chimeric primer that can be detected and can specifically hybridize to a gene, and a DNA having strand displacement activity. Including polymerase and endonuclease RNase.
- the internal control is used as an internal standard for monitoring amplification or quantification of the target nucleic acid (DNA, RNA).
- the sequence of the internal control has the same primer as the sample primer on both sides of the sequence different from the sample, so that it can be amplified similarly to the sample.
- the sequence of the positive control is a specific sequence for detecting a sample, and the portion between the primer and the hybridized portion and the sequence between them are the same as the sample.
- Nucleic acid used for control (DN A, RNA) may be those described in known technical literature.
- Negative controls include all reagents other than nucleic acids (DNA, RNA), and are used to check for contamination and to correct for knock ground.
- RNA sample it is a reverse transcriptase or reverse transcription primer for synthesizing cDNA from RNA, and these are also commercially available and easily available.
- amplification substrates (2'-deoxynucleoside 5'-triphosphate), gene amplification reagents, and the like are preliminarily sealed in predetermined amounts in the reagent storage sections of one microreactor. . Therefore, the microreactor of the present invention can be used immediately without using a required amount of reagent each time it is used.
- the method for detecting the DNA of the target gene amplified in the present invention is not particularly limited, and a suitable method is used if necessary. Among such methods, detection methods such as visible light spectroscopy, fluorescence measurement, and luminescence are mainly used. In addition, there are other methods such as electrochemical method, surface plasmon resonance, and quartz crystal microbalance.
- the gene testing apparatus of the present invention together with the microreactor, performs detection for detecting the presence or absence, scale, and the like of an amplification reaction based on a reactive product by hybridization of the amplified gene and probe DNA.
- the method of the present invention using the microreactor is specifically performed in the following steps. That is, using the microreactor described above, (1) the DNA extracted from the sample or the sample power or the cDNA synthesized by reverse transcription reaction from the extracted RNA from the sample or the sample power and the primer modified with biotin are transferred from these storage sections to the flow channel. Sending the solution and amplifying the gene in the microchannel, (2) mixing the amplification reaction solution containing the gene amplified in the microchannel and the denaturing solution, and converting the amplified gene into a single strand.
- a step of denaturation treatment (3) a treatment solution in which the amplified gene is denatured into a single strand is sent into a microchannel to which streptavidin is adsorbed, and the amplified gene is immobilized.
- Process (4) Fluorescein isothiocyanate (FITC) at the end of the microchannel in which the amplified gene is immobilized.
- FITC Fluorescein isothiocyanate
- a step of sending a washing liquid into the flow path to which streptavidin has been adsorbed, if necessary, between the above steps is included.
- a washing solution for example, various buffers, aqueous saline solutions, organic solvents and the like are suitable.
- the detection method of the present invention is preferably a method that can be measured with high sensitivity by finally using visible light. Compared to fluorescence photometry, the equipment is more versatile, has fewer interfering factors, and data processing is easier.
- the optical detection device for that purpose is integrated with a liquid sending means including the micropump and a temperature control device for controlling the reaction temperature of each reaction in the flow path of the microreactor, and has an integrated configuration. The detection is performed using the genetic test apparatus of the present invention.
- the denaturing solution is a reagent for converting the gene DNA into a single strand, and examples thereof include sodium hydroxide and potassium hydroxide.
- the probe include oligodeoxynucleotides.
- FITC fluorescence tomography
- RITC rhodamine isothiocynate
- the above amplification and detection are performed by controlling various conditions set in advance with respect to the liquid sending order, volume, timing, and the like as well as the control of the micropump and the temperature as the contents of the program.
- the flow path of the microreactor is also activated.
- the sample injection preferably initiates the analysis automatically, and the sample and reagent transfer, the gene amplification reaction based on mixing, the gene detection reaction and the optical measurement are performed automatically as a series of continuous steps.
- the measurement data is stored in a file together with necessary conditions and recorded items.
- the presence or absence of amplification or the amplification efficiency can be measured to determine the DNA power derived from the gene in the sample. It can be used to determine whether they have the same power as the gene or whether they are different. In particular, it is effective for quickly identifying the causative virus or bacterium of an infectious disease from a gene.
- Data for diagnosing the degree of expression of the oncogene, the hereditary hypertension gene, and the like can be obtained by the genetic test of the present invention. Specifically, it is an analysis of the type and expression level of mRNA, which is a proof of the expression of such a gene.
- mutations in the promoter region of regulatory genes can also be detected by genetic testing using the microreactor of the present invention.
- a primer having a nucleic acid sequence containing a mutated portion is used.
- the above gene mutation means a mutation at a nucleotide base of a gene.
- the analysis of a gene polymorphism by using the genetic test device of the present invention is also useful for identifying a disease susceptibility gene.
- the genetic test method using the genetic test device of the present invention uses a much smaller sample amount, a small amount of labor, and a simple device as compared with conventional nucleic acid sequence analysis, restriction enzyme analysis, and nucleic acid hybridization analysis. It is clear from the configuration of the device and the principle of analysis that high-accuracy results can be obtained.
- the microreactor for genetic testing, the apparatus for genetic testing, and the like of the present invention include gene expression analysis, gene function analysis, single gene polymorphism analysis (SNP), clinical testing, diagnosis, pharmaceutical screening, pharmaceuticals, agricultural chemicals, and the like. It can be used in the fields of safety of various chemicals, toxicity testing, environmental analysis, food testing, forensic medicine, chemistry, brewing, fisheries, livestock, agricultural production, agriculture and forestry.
- SNP single gene polymorphism analysis
- FIG. 1 is a schematic diagram of a microreactor for genetic testing according to one embodiment of the present invention
- FIG. 2 is a schematic diagram of a genetic testing device including the microreactor and a device main body.
- the microreactor shown in Fig. 1 is composed of a single resin chip, and is used to transfer a sample such as blood.
- a sample such as blood.
- gene amplification reaction and its detection are automatically performed in the chip, and gene diagnosis can be performed simultaneously for multiple items. For example, by simply dropping a blood sample of about 2 to 3 ⁇ l on a chip that is several cm in length and width, the amplification reaction and its detection can be performed by attaching the chip to the device body 2 in Fig. 2. ing.
- the sample injected into the sample storage unit 20 of FIG. 1 and the reagent used for the gene amplification reaction previously sealed in the reagent storage units 18a to 18c are provided by a micropump (shown in FIG. ),
- the liquid is sent to the flow path communicating with each storage section, the sample and the reagent are mixed in the flow path via the Y-shaped flow path, and the amplification reaction is performed.
- the flow path is formed to have a width of about 100 m and a depth of about 100 m, for example, and the amplification reaction is detected by an optical detection device (not shown) incorporated in the device main body 2 of FIG.
- the probe DNA is hybridized by irradiating measurement light from an LED to the flow path for each inspection item and detecting transmitted light or reflected light by light detection means such as a photodiode or a photomultiplier tube.
- light detection means such as a photodiode or a photomultiplier tube.
- the apparatus main body 2 also incorporates a temperature control device for controlling the reaction temperature, and the reagent is previously sealed in a small unit in which the liquid sending pump, the optical detection device, and the temperature control device are integrated.
- Gene diagnosis can be easily performed simply by attaching the chip. In this way, measurement can be performed quickly regardless of location and time, so that it can be used for emergency medical care and personal use for home medical care. Since a large number of micropump units and the like used for liquid transfer are incorporated in the main body of the apparatus, the chip can be used as a disposable type.
- the microreactor of the present embodiment preferably performs an amplification reaction by the ICAN method, and in the microreactor, a sample extracted from blood or sputum power and a biotin modification that specifically hybridizes to a gene to be detected.
- a gene amplification reaction is carried out using the thus obtained chimeric primer, a DNA polymerase having strand displacement activity, and a reagent containing endonuclease. After the denaturation treatment, the reaction solution is sent to a channel in which streptavidin is adsorbed, and the amplified gene is fixed in the channel.
- the probe DNA modified with TC and the immobilized gene are hybridized, and the gold colloid whose surface has been modified with the FITC antibody is adsorbed to the probe hybridized to the immobilized gene, and the gold colloid is absorbed.
- the amplified gene is detected by optically measuring the concentration of the gene.
- a microreactor is configured as follows in order to perform a highly accurate, quick and highly reliable genetic test with one chip. First, all controls are integrated into one chip, and the internal control, positive control, and negative control are pre-enclosed in a microreactor. The amplification reaction and detection operation are performed. As a result, the genetic test can be performed quickly and with high reliability.
- the passage of the liquid is blocked at each position of the flow path until the liquid sending pressure in the forward direction reaches a preset pressure, and a liquid sending pressure higher than the preset pressure is applied.
- a liquid sending control unit that allows the passage of the liquid by the pump pressure of the micropump, and a backflow prevention unit that prevents the backflow of the liquid in the flow path are provided.
- the micropump, the liquid sending control unit, and the backflow prevention unit control the liquid sending in the flow channel, and can send a fixed amount of reagent and the like with high precision. A plurality of reagents can be quickly mixed.
- the microreactor is provided with a plurality of reagent containers for accommodating each reagent, the reagent used for the gene amplification reaction, the denaturing solution for denaturing the amplified gene, and the hybridization between the amplified gene and the hybridization.
- the probe DNA to be accommodated is accommodated.
- a reagent is previously stored in the reagent storage section so that the test can be performed promptly regardless of location or time.
- the surface of the reagent section of the reagents and the like built in the chip is hermetically sealed in order to prevent evaporation, leakage, mixing of air bubbles, contamination, and denaturation.
- the microreactor is stored, it is sealed with a sealing material in order to prevent the reagent from leaking into the fine flow channel and reacting with the reagent.
- These sealants are solidified or gelled under refrigerated conditions under which ⁇ -TAS (microreactor) is stored before use, and when used, they melt and become a fluid state at room temperature. is there. As shown in FIG.
- the reagent is sealed in the reagent container by filling a sealant 32 between the reagent 31 and the channel 15 communicating with the reagent container 18. Air may be interposed between the sealant and the reagent, but it is preferable that the amount of air interposed between the sealant and the reagent is sufficiently small (relative to the amount of the reagent). .
- a plastic material having low solubility in water can be used, and an oil or fat having a solubility in water of 1% or less is preferable.
- Such fats and oils can be examined with a fats and oils handbook and the like, and for example, the fats and oils shown in Table 1 can be mentioned.
- reagents are stored in the microreactor in advance, it is desirable to keep the microreactors refrigerated in view of the stability of the reagents.
- a substance that is in a solid state during refrigeration and becomes liquid at room temperature is used as a sealant.
- the reagent can be sealed in a solid state at the time of refrigerated storage, and can be discharged in a liquid state at the time of use and can be easily discharged.
- sealing agents include oils and fats having a solubility in water of 1% or less and a melting point of 8 ° C. to room temperature (25 ° C.), and an aqueous solution of gelatin.
- the gelling temperature of the aqueous gelatin solution can be adjusted by changing the concentration of the gelatin. For example, in order to gel at about 10 ° C., an approximately 1% aqueous solution may be used.
- the sealant may be similarly filled between each of the accommodating sections accommodating the positive control and the negative control and the flow path communicating therewith.
- a micropump is connected to the upstream side of the reagent storage section, and the driving liquid is supplied to the reagent storage section side by the micropump, whereby the reagent is pushed out to the flow path and sent. .
- a micropump is provided for each of the sample storage unit, the reagent storage unit, the positive control storage unit, and the negative control storage unit, for supplying the liquid in these storage units.
- the micropump is built into the main body of the device separately from the microreactor.
- the pump connection force is also connected to the microreactor!
- FIG. 4 (a) is a cross-sectional view showing an example of this pump
- FIG. 4 (b) is a top view thereof.
- the micro pump includes a substrate 42 having a first liquid chamber 48, a first flow path 46, a pressurizing chamber 45, a second flow path 47, and a second liquid chamber 49, and is laminated on the substrate 42.
- a drive unit (not shown) for driving the element 44 is provided.
- a 500 ⁇ m-thick photosensitive glass substrate is used as the substrate 42, and etching is performed until the depth reaches 100 ⁇ m, whereby the first liquid chamber 48, the first flow path 46, A pressurizing chamber 45, a second flow path 47 and a second liquid chamber 49 are formed.
- the first channel 46 has a width of 25 / ⁇ and a length of 20 m.
- the second channel 47 has a width of 25 m and a length of 150 m.
- the upper substrate 41 which is a glass substrate
- the upper surfaces of the first liquid chamber 48, the first flow path 46, the second liquid chamber 49, and the second flow path 47 are formed.
- the portion of the upper substrate 41 corresponding to the upper surface of the pressurizing chamber 45 is processed by etching or the like and penetrates.
- a vibrating plate 43 having a thickness of 50 ⁇ m and also having a thin glass force is laminated, and a force such as a lead zirconate titanate (PZT) ceramic having a thickness of 50 m is formed thereon.
- PZT lead zirconate titanate
- the piezoelectric element 44 and the vibrating plate 43 attached to the piezoelectric element 44 vibrate due to the driving voltage of the driving unit, whereby the volume of the pressurizing chamber 45 increases or decreases.
- the first flow path 46 and the second flow path 47 have the same width and depth, and the length of the second flow path is longer than that of the first flow path.
- the differential pressure increases, a turbulent flow occurs in a swirl in the flow path, and the flow resistance increases.
- the second flow path 47 since the flow path width is long, even if the differential pressure is large, the rate of change in the flow path resistance with respect to the change in the differential pressure is smaller than in the first flow path, which tends to be laminar.
- the driving voltage applied to the piezoelectric element 44 causes the diaphragm 43 to be quickly displaced inward of the pressurizing chamber 45 to reduce the volume of the pressurizing chamber 45 while giving a large pressure and a differential pressure.
- the volume of the pressure chamber 45 is increased while slowly displacing the vibration plate 43 outward from the pressure chamber 45 to apply a small pressure difference, the liquid is sent in the direction B in FIG.
- the diaphragm 43 is quickly displaced outward from the pressurizing chamber 45 to increase the volume of the pressurizing chamber 45 while applying a large differential pressure, and then the diaphragm 43 is slowly moved inward from the pressurizing chamber 45.
- FIG. 5 shows an example of the relationship between the drive voltage waveform applied to the piezoelectric element 44 and the displacement of the liquid.
- the graph of the amount of liquid movement shown in Fig. 5 (b) schematically shows the flow rate obtained by the pump operation. Vibration is superimposed.
- the difference in the change ratio of the flow path resistance with respect to the change in the differential pressure between the first flow path and the second flow path is based on other geometrical differences that are not necessarily required due to the difference in the flow path length. It may be something.
- the piezo pump configured as described above, by changing the driving voltage and frequency of the pump, it is possible to control the liquid sending direction and the liquid sending speed.
- Fig. 4 (c) shows another example of this pump.
- the pump has a silicon substrate 71, a piezoelectric element 44, and a flexible wiring force (not shown).
- the silicon substrate 71 is obtained by processing a silicon wafer into a predetermined shape by a known photolithography technique.
- a pressure chamber 45, a diaphragm 43, a first flow path 46, a first liquid chamber 48, a second flow path 47, and a second liquid chamber 49 are formed by etching.
- the first liquid chamber 48 is provided with a port 72
- the second liquid chamber 49 is provided with a port 73, and communicates with the pump connection portion of the microreactor via this port.
- the pump can be connected to the microreactor by vertically stacking the substrate 74 with the perforated port and the vicinity of the pump connection portion of the microreactor.
- a plurality of pumps can be formed on one silicon substrate. In this case, it is desirable to connect a driving liquid tank to the port on the opposite side of the port connected to the microreactor. If there are multiple pumps, those ports may be connected to a common drive fluid tank.
- FIG. 6 shows the configuration around the pump connection part.
- FIG. 1A shows the configuration of a pump unit that sends a driving liquid
- FIG. 1B shows the configuration of a pump unit that sends a reagent.
- the driving liquid 24 may be oil-based such as mineral oil or water-based.
- the sealing liquid 25 for sealing the reagent may be filled in the flow path as shown in FIG. Alternatively, it may be filled in a storage section provided for a sealing liquid.
- a flow path 26 for air release is provided in the flow path between the pump connection section 12 and the reagent storage section 18.
- the air vent channel 26 branches off from the channel 15 between the pump connection part and the reagent storage part, and its terminal is open. Air bubbles present in the flow path 15 are removed from the air release flow path 26 when, for example, a pump is connected.
- the air vent channel 26 has a channel diameter of 10 m or less and prevents the aqueous liquid 27 such as water from passing through the channel 15 from leaking out.
- the contact angle is 30 ° or more.
- each micropump that sends these is controlled as follows.
- the reagent 31 is sent in the direction A and the sample 33 is sent in the direction B from the upstream of the Y-shaped branch flow path, so that they are mixed in the flow path 15.
- the drive of the pump for sending the reagent 31 and the drive of the pump for sending the sample 33 are controlled as shown in FIG. 8C.
- the feeding of the sample 33 is stopped while the reagent 31 is sent in the direction A, and the feeding of the reagent 31 is stopped while the sample 33 is sent in the direction B.
- the reagent 31 and the sample 33 are alternately filled in the channel 15 in a ring shape as shown in FIG. 8A.
- the width of the slice layer can be set to 1 to 2 m. The shorter the width of the layer, the faster the diffusion between the reagent 31 and the sample 33 proceeds and the faster the mixing.
- reagent 31 and sample 33 are sent to channel 15 at a fixed ratio of 1: 1 in a channel with a channel diameter of 100 m, as shown in Fig. 8 (b), approximately 50 ⁇ m A reagent layer having a width and a sample layer are formed, and the mixing proceeds more slowly as compared with the case of FIG. 8 (a).
- the microreactor of the present embodiment is provided with a number of liquid sending controllers as shown in FIG.
- the liquid supply control unit blocks the passage of the liquid until the liquid supply pressure in the forward direction reaches the predetermined pressure, and allows the liquid to pass through by applying a liquid supply pressure equal to or higher than the predetermined pressure.
- the liquid sending control section 13 is formed of a portion having a reduced flow path diameter, and this allows the narrowed flow path (narrow flow path) 51 to be connected from one end side. Restricts the liquid that has reached to the other end.
- the throttle channel 51 has, for example, a length of 150 It is formed so that the length and width are about 30 mx 30 m for a flow path of mx 150 m.
- a water-repellent coating for example, a fluorine-based coating may be applied to the inner surface of the throttle channel 51.
- the cross-sectional area formed between these flow paths so as to connect the flow paths adjacent on both sides in series and perpendicular to the flow path axis direction in these adjacent flow paths is smaller than the cross-sectional area.
- the microreactor of the present embodiment is provided with a large number of backflow prevention units for preventing backflow of liquid in its flow path.
- the backflow prevention unit is a check valve in which the valve body closes the flow path opening by the backflow pressure, or an active valve that closes the opening by pressing the valve body to the flow path opening by the valve body deformation means. Power.
- FIGS. 10 (a) and 10 (b) are cross-sectional views showing an example of a check valve used for the flow channel of the microreactor of the present embodiment.
- the check valve of FIG. 10A the passage of the liquid is allowed and blocked by opening and closing the opening 68 formed in the substrate 62 by the movement of the microsphere 67 using the microsphere 67 as a valve element. That is, when the liquid is sent from the direction A, the microsphere 67 is separated from the substrate 62 by the liquid pressure and the opening 68 is opened, so that the passage of the liquid is allowed. On the other hand, when the liquid flows backward from the direction B, the microsphere 67 is seated on the substrate 62 and the opening 68 is closed, so that the passage of the liquid is blocked.
- a flexible substrate 69 laminated on the substrate 62 and having its end extending above the opening 68 moves up and down above the opening 68 by hydraulic pressure. Opening 68 is opened and closed.
- the flexible substrate 69 comes into close contact with the substrate 62 and the opening 68 is closed, so that the passage of the liquid is blocked.
- FIG. 11 is a cross-sectional view showing an example of the active valve used in the flow channel of the microreactor of the present embodiment.
- FIG. 11 (a) shows the valve in an open state
- FIG. 11 (b) shows the valve closed. Indicates the status.
- a flexible substrate 63 having a valve portion 64 protruding downward is formed on a substrate 62 having an opening 65 formed thereon.
- the flexible substrate 63 is also pressed upward by a valve body deforming means such as a pneumatic, hydraulic, or hydraulic piston, a piezoelectric actuator, or a shape memory alloy actuator.
- a valve body deforming means such as a pneumatic, hydraulic, or hydraulic piston, a piezoelectric actuator, or a shape memory alloy actuator.
- the valve portion 64 is brought into close contact with the substrate 62 so as to cover the opening 65, thereby blocking the backflow in the B direction.
- the active valve is not limited to one that is operated by an external driving device, and may have a configuration in which the valve body deforms itself to close the flow path. For example, as shown in FIG. 18, it may be deformed by electric heating using a bimetal 81, or as shown in FIG. 19, may be deformed by electric heating using a shape memory alloy 82. Just a little.
- FIG. 12 is a diagram showing the configuration of such a reagent quantification unit.
- a predetermined amount of reagent is provided in a flow path (reagent filling flow path 15a) between the backflow prevention unit 16 and the liquid sending control unit 13a. Is filled.
- a branch channel 15b is provided, which branches off from the reagent filling channel 15a and communicates with the micropump 11 that sends the driving liquid.
- the fixed-quantity liquid sending of the reagent is performed as follows. First, the reagent 31 is filled by supplying the reagent 31 to the reagent filling channel 15a at the liquid sending pressure from the backflow prevention unit 16 side without the reagent 31 passing from the liquid sending control unit 13a. Next, the micro pump 11 sends the driving liquid 25 from the branch flow path 15b to the reagent filling flow path 15a in the direction of the force at a liquid sending pressure that allows the reagent 31 to pass from the liquid sending control unit 13a. As a result, the reagent 31 filled in the reagent filling flow path 15a is pushed out from the liquid sending control unit 15a, whereby the reagent 31 is quantitatively sent.
- the reagent can be pushed out by sending the driving liquid 25 by the micropump 11 and sending the air, the sealing liquid and the like into the reagent filling channel 15a.
- FIG. 13 is a diagram showing a channel configuration in which the head portion is truncated so that the mixture ratio is stabilized and the mixture is sent to the next step.
- the reagents 31a and 31b to be mixed are sent from the channels 15a and 15b to the mixing channel 15c, respectively.
- a branch flow path 15d for sending the reagent mixture 31c to the next step is branched, and a position before the branch point of the mixing flow path 15c with the branch flow path 15d is provided.
- the first liquid supply control unit 13a is provided.
- the liquid transfer pressure at which the reagent mixture 31c can pass is smaller than that of the first liquid transfer control unit 13a, and the second liquid transfer control is performed.
- a part 13b is provided.
- the reagent mixture 31c of the reagent 31a and the reagent 31b sent from the flow paths 15a and 15b into the mixing flow path 15c has its leading end 31d reaching the first liquid sending control section 13a. Is fed through the mixed channel 15c. After the leading end 31d of the reagent mixture 31c reaches the first liquid supply controller 13a, the mixture is further supplied into 15c, so that the reagent mixture 31b flows from the second liquid supply controller 13b to the branch channel 15d. Then, 3 lc of the reagent mixture is sent to the next step.
- the second liquid flow control For example, by making the cross-sectional area of the narrow flow path in the first liquid flow control unit 13a smaller than the cross-sectional area of the fine flow path in the second liquid flow control unit 13b, the second liquid flow control The liquid sending pressure at which the reagent mixture 31c can pass through the section 13b can be reduced by J / J from that of the first liquid sending control section 13a.
- FIGS. 14 to 17 Biotin-specific primers that hybridize specifically to the gene to be detected, DNA polymerases with strand displacement activity, and endonucleases.
- Which reagents are stored in the reagent storage sections 18a, 18b, and 18c in Fig. 14, and a piezo pump 11 built in the main body of the apparatus separate from the microreactor is connected to the upstream side of each reagent storage section by a pump connection section 12. Then, the reagents are sent from these reagent storage sections to the downstream flow path 15a by these pumps.
- the flow path 15a, the flow path from the flow path 15a to the next process branched from the flow path 15a, and the liquid sending control sections 13a and 13b constitute the flow paths described with reference to Fig. 13, and the liquid is sent from each reagent storage section.
- the tip of the mixed solution of the reagent is cut off, and the mixed solution is sent to the next process after the mixed state is stabilized.
- Each reagent storage section contains a total of more than 7.51 reagents, and a total of 7.51 reagent mixtures, truncated at the tip, are divided into three streams of 2.51 each.
- the liquid is sent to channels 15b, 15c and 15d.
- Channel 15b is for reaction with analyte and detection system (Figure 15)
- Channel 15c is for reaction with positive control and detection system (Figure 16)
- Channel 15d is for reaction with negative control and detection system ( Figure 16). (Fig. 17).
- the mixed reagent sent to the flow path 15b is filled in the storage unit 17 in FIG.
- the reagent filling flow path described with reference to FIG. 12 is configured between the check valve 16 on the upstream side of the storage section 17a and the liquid sending control section 13a on the downstream side, and the pump 11 for sending the driving liquid is provided. Together with the liquid sending control unit 13b provided in the communicating branch flow path, it constitutes the above-described reagent quantitative unit.
- the sample from which the blood or sputum power is also extracted is injected from the sample storage unit 20, and the storage unit 17b is filled with the sample by the same mechanism as the reagent quantification unit described above (2.5 ⁇ 1), followed by The fixed amount is sent to the channel.
- the sample and the reagent mixture filled in each of the reservoirs 17a and 17b are sent to the channel 15e (volume 5 ⁇ l) via the Y-shaped channel, where the mixing and the ICAN reaction are performed. Done.
- the pumps 11 are alternately driven to introduce the sample and the reagent mixture alternately into the channel 15e in a ring shape as described in FIG.
- the body and the reagent are allowed to diffuse and mix.
- the probe DNA solution (2.51) whose terminal was fluorescently labeled with FITC and the denatured treatment solution (1.51) accommodated in the probe DNA accommodation section 21c were placed in a volume 4 volume.
- the solution is sent to the channel 15h of 1 and mixed, and the probe DNA is hybridized to the single-stranded amplified gene.
- the washing solution, the internal control probe DNA solution, and the FITC antibody contained in each of the accommodating parts 21d, 21f, and 21e were introduced into the channel 22a in which the amplified gene was immobilized by a single pump 11.
- the solution of gold colloid labeled with is sent in the order shown in FIG.
- a single pump 11 is used to label the amplified gene in the channel 22b where the amplified gene is immobilized, with the washing solution, the probe DNA solution for MTB, and the FIT C antibody contained in each of the reservoirs 21d, 21g, and 21e.
- the gold colloid solution is sent in the order shown in FIG.
- the colloidal gold is bound to the immobilized amplified gene via FITC and fixed.
- the presence or absence of amplification or amplification efficiency is measured by optically detecting the immobilized gold colloid.
- the flow paths 15c and 15d in Fig. 14 are connected to the positive control reaction and detection system shown in Fig. 16 and the negative control reaction and detection system shown in Fig. 17, respectively.
- the amplification reaction is performed in the flow channel with the reagent, and then the probe DNA stored in the probe DNA storage is hybridized in the flow channel. The amplification reaction is detected based on the reaction product.
Abstract
Description
Claims
Priority Applications (2)
Application Number | Priority Date | Filing Date | Title |
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EP05737310A EP1746158A4 (en) | 2004-05-07 | 2005-04-27 | MICROREACTOR FOR TESTING, GENETIC TEST EQUIPMENT AND GENETIC TEST PROCEDURE |
JP2006512967A JP4784508B2 (ja) | 2004-05-07 | 2005-04-27 | 検査用マイクロリアクタおよび検査装置ならびに検査方法 |
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JP2004-138959 | 2004-05-07 | ||
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US (1) | US7906318B2 (ja) |
EP (1) | EP1746158A4 (ja) |
JP (1) | JP4784508B2 (ja) |
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WO2007080850A1 (ja) * | 2006-01-12 | 2007-07-19 | Sumitomo Bakelite Co., Ltd. | 受動型一方弁及びマイクロ流体デバイス |
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WO2007145040A1 (ja) * | 2006-06-12 | 2007-12-21 | Konica Minolta Medical & Graphic, Inc. | 液漏れ防止機構を備えたマイクロ総合分析システム |
JP2008012490A (ja) * | 2006-07-07 | 2008-01-24 | Shimadzu Corp | 微量化学反応方法及び装置 |
JP2008511842A (ja) * | 2004-09-02 | 2008-04-17 | ハネウェル・インターナショナル・インコーポレーテッド | 微小流体回路の1つまたは複数の動作パラメータを決定する方法と装置 |
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WO2008090760A1 (ja) * | 2007-01-26 | 2008-07-31 | Konica Minolta Medical & Graphic, Inc. | 蛍光検出装置、マイクロチップ、及び検査システム |
JP2009145105A (ja) * | 2007-12-12 | 2009-07-02 | Konica Minolta Medical & Graphic Inc | マイクロチップ |
JP2009210327A (ja) * | 2008-03-03 | 2009-09-17 | Yokogawa Electric Corp | 化学反応用カートリッジ、混合物生成方法及び化学反応用カートリッジの制御装置 |
US8067176B2 (en) | 2008-01-25 | 2011-11-29 | Shimadzu Corporation | Microchemistry reaction method |
JP2013167641A (ja) * | 2007-07-13 | 2013-08-29 | Handylab Inc | マイクロ流体カートリッジにおいて核酸増幅を行う装置及び方法 |
US8845980B2 (en) | 2007-09-10 | 2014-09-30 | Nec Corporation | Sample packing device |
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US9480983B2 (en) | 2011-09-30 | 2016-11-01 | Becton, Dickinson And Company | Unitized reagent strip |
US9528142B2 (en) | 2001-02-14 | 2016-12-27 | Handylab, Inc. | Heat-reduction methods and systems related to microfluidic devices |
US9618139B2 (en) | 2007-07-13 | 2017-04-11 | Handylab, Inc. | Integrated heater and magnetic separator |
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US9670528B2 (en) | 2003-07-31 | 2017-06-06 | Handylab, Inc. | Processing particle-containing samples |
US9677121B2 (en) | 2001-03-28 | 2017-06-13 | Handylab, Inc. | Systems and methods for thermal actuation of microfluidic devices |
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US9802199B2 (en) | 2006-03-24 | 2017-10-31 | Handylab, Inc. | Fluorescence detector for microfluidic diagnostic system |
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US10100302B2 (en) | 2007-07-13 | 2018-10-16 | Handylab, Inc. | Polynucleotide capture materials, and methods of using same |
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US10179910B2 (en) | 2007-07-13 | 2019-01-15 | Handylab, Inc. | Rack for sample tubes and reagent holders |
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US10364456B2 (en) | 2004-05-03 | 2019-07-30 | Handylab, Inc. | Method for processing polynucleotide-containing samples |
US10571935B2 (en) | 2001-03-28 | 2020-02-25 | Handylab, Inc. | Methods and systems for control of general purpose microfluidic devices |
US10799862B2 (en) | 2006-03-24 | 2020-10-13 | Handylab, Inc. | Integrated system for processing microfluidic samples, and method of using same |
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US10900066B2 (en) | 2006-03-24 | 2021-01-26 | Handylab, Inc. | Microfluidic system for amplifying and detecting polynucleotides in parallel |
JP2021081449A (ja) * | 2013-03-16 | 2021-05-27 | ドン ロバーツ レスリー | 自己完結型モジュラー分析カートリッジ及びプログラム可能試薬送達システム |
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US11453906B2 (en) | 2011-11-04 | 2022-09-27 | Handylab, Inc. | Multiplexed diagnostic detection apparatus and methods |
US11806718B2 (en) | 2006-03-24 | 2023-11-07 | Handylab, Inc. | Fluorescence detector for microfluidic diagnostic system |
US11865544B2 (en) | 2013-03-15 | 2024-01-09 | Becton, Dickinson And Company | Process tube and carrier tray |
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Families Citing this family (21)
Publication number | Priority date | Publication date | Assignee | Title |
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Citations (12)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JPH06259138A (ja) * | 1993-03-02 | 1994-09-16 | Toshiba Corp | 流量制御装置 |
WO1996006897A2 (de) * | 1994-08-31 | 1996-03-07 | Henkel Kommanditgesellschaft Auf Aktien | Physikalisch und/oder chemisch abbindende bindemittel |
JPH1080414A (ja) * | 1996-06-04 | 1998-03-31 | Arukea Kk | Mri用マーカー剤及びマーカー |
JPH10337173A (ja) * | 1997-06-05 | 1998-12-22 | Rikagaku Kenkyusho | 生化学反応用マイクロリアクタ |
JP2000081154A (ja) * | 1998-09-03 | 2000-03-21 | Hitachi Ltd | マイクロバルブ |
WO2000037163A1 (en) * | 1998-12-23 | 2000-06-29 | Nanogen, Inc. | Integrated portable biological detection system |
JP2000266337A (ja) * | 1999-03-12 | 2000-09-29 | Babcock Hitachi Kk | 固体・水混合燃料の供給装置および供給方法 |
JP2001029848A (ja) * | 1999-07-19 | 2001-02-06 | Honda Motor Co Ltd | 粉体塗料の塗装ガンへの供給装置 |
US20020009374A1 (en) * | 2000-05-16 | 2002-01-24 | Kusunoki Higashino | Micro pump |
US20020064483A1 (en) * | 2000-11-20 | 2002-05-30 | Yasuhiro Sando | Microchip |
US20020071788A1 (en) * | 2000-12-08 | 2002-06-13 | Minolta Co., Ltd. | Microchip |
WO2003038436A2 (en) * | 2001-11-02 | 2003-05-08 | University Of Strathclyde | Microfluidic ser(r)s detection |
Family Cites Families (14)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US5856174A (en) * | 1995-06-29 | 1999-01-05 | Affymetrix, Inc. | Integrated nucleic acid diagnostic device |
US6130098A (en) * | 1995-09-15 | 2000-10-10 | The Regents Of The University Of Michigan | Moving microdroplets |
US20020068357A1 (en) * | 1995-09-28 | 2002-06-06 | Mathies Richard A. | Miniaturized integrated nucleic acid processing and analysis device and method |
US5958344A (en) * | 1995-11-09 | 1999-09-28 | Sarnoff Corporation | System for liquid distribution |
NZ333346A (en) * | 1996-06-28 | 2000-03-27 | Caliper Techn Corp | High-throughput screening assay systems in microscale fluidic devices |
US6235471B1 (en) * | 1997-04-04 | 2001-05-22 | Caliper Technologies Corp. | Closed-loop biochemical analyzers |
US6637463B1 (en) * | 1998-10-13 | 2003-10-28 | Biomicro Systems, Inc. | Multi-channel microfluidic system design with balanced fluid flow distribution |
US20040053290A1 (en) * | 2000-01-11 | 2004-03-18 | Terbrueggen Robert Henry | Devices and methods for biochip multiplexing |
US6706519B1 (en) * | 1999-06-22 | 2004-03-16 | Tecan Trading Ag | Devices and methods for the performance of miniaturized in vitro amplification assays |
US20020187564A1 (en) * | 2001-06-08 | 2002-12-12 | Caliper Technologies Corp. | Microfluidic library analysis |
CN1258602C (zh) * | 2001-09-22 | 2006-06-07 | 香港基因晶片开发有限公司 | 一种侦测基因序列的装置和方法 |
US7455770B2 (en) * | 2002-09-09 | 2008-11-25 | Cytonome, Inc. | Implementation of microfluidic components in a microfluidic system |
EP1654347B1 (en) * | 2003-06-26 | 2014-06-04 | Seng Enterprises Limited | Improved materials for constructing cell-chips, cell-chip covers, cell-chip coats, processed cell-chips and uses thereof |
JP4996248B2 (ja) * | 2003-07-31 | 2012-08-08 | ハンディーラブ インコーポレイテッド | 粒子含有サンプルの処理 |
-
2005
- 2005-04-27 CN CNB2005800141047A patent/CN100516212C/zh not_active Expired - Fee Related
- 2005-04-27 EP EP05737310A patent/EP1746158A4/en not_active Withdrawn
- 2005-04-27 WO PCT/JP2005/008051 patent/WO2005108571A1/ja not_active Application Discontinuation
- 2005-04-27 JP JP2006512967A patent/JP4784508B2/ja not_active Expired - Fee Related
- 2005-05-04 US US11/121,096 patent/US7906318B2/en active Active
Patent Citations (12)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JPH06259138A (ja) * | 1993-03-02 | 1994-09-16 | Toshiba Corp | 流量制御装置 |
WO1996006897A2 (de) * | 1994-08-31 | 1996-03-07 | Henkel Kommanditgesellschaft Auf Aktien | Physikalisch und/oder chemisch abbindende bindemittel |
JPH1080414A (ja) * | 1996-06-04 | 1998-03-31 | Arukea Kk | Mri用マーカー剤及びマーカー |
JPH10337173A (ja) * | 1997-06-05 | 1998-12-22 | Rikagaku Kenkyusho | 生化学反応用マイクロリアクタ |
JP2000081154A (ja) * | 1998-09-03 | 2000-03-21 | Hitachi Ltd | マイクロバルブ |
WO2000037163A1 (en) * | 1998-12-23 | 2000-06-29 | Nanogen, Inc. | Integrated portable biological detection system |
JP2000266337A (ja) * | 1999-03-12 | 2000-09-29 | Babcock Hitachi Kk | 固体・水混合燃料の供給装置および供給方法 |
JP2001029848A (ja) * | 1999-07-19 | 2001-02-06 | Honda Motor Co Ltd | 粉体塗料の塗装ガンへの供給装置 |
US20020009374A1 (en) * | 2000-05-16 | 2002-01-24 | Kusunoki Higashino | Micro pump |
US20020064483A1 (en) * | 2000-11-20 | 2002-05-30 | Yasuhiro Sando | Microchip |
US20020071788A1 (en) * | 2000-12-08 | 2002-06-13 | Minolta Co., Ltd. | Microchip |
WO2003038436A2 (en) * | 2001-11-02 | 2003-05-08 | University Of Strathclyde | Microfluidic ser(r)s detection |
Non-Patent Citations (2)
Title |
---|
KITAMORI ET AL: "Kagaku Kenkyu ha Microchip no Uede", CHEMISTRY, vol. 54, no. 10, 1999, pages 14 - 19, XP002994428 * |
See also references of EP1746158A4 * |
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US8257974B2 (en) | 2006-03-29 | 2012-09-04 | Konica Minolta Medical & Graphic, Inc. | Method of reaction in flow channel of microchip and analysis device |
WO2007145040A1 (ja) * | 2006-06-12 | 2007-12-21 | Konica Minolta Medical & Graphic, Inc. | 液漏れ防止機構を備えたマイクロ総合分析システム |
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US10234474B2 (en) | 2007-07-13 | 2019-03-19 | Handylab, Inc. | Automated pipetting apparatus having a combined liquid pump and pipette head system |
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US10844368B2 (en) | 2007-07-13 | 2020-11-24 | Handylab, Inc. | Diagnostic apparatus to extract nucleic acids including a magnetic assembly and a heater assembly |
US10139012B2 (en) | 2007-07-13 | 2018-11-27 | Handylab, Inc. | Integrated heater and magnetic separator |
US11254927B2 (en) | 2007-07-13 | 2022-02-22 | Handylab, Inc. | Polynucleotide capture materials, and systems using same |
JP2016000054A (ja) * | 2007-07-13 | 2016-01-07 | ハンディーラブ インコーポレイテッド | 診断装置 |
US10875022B2 (en) | 2007-07-13 | 2020-12-29 | Handylab, Inc. | Integrated apparatus for performing nucleic acid extraction and diagnostic testing on multiple biological samples |
US10100302B2 (en) | 2007-07-13 | 2018-10-16 | Handylab, Inc. | Polynucleotide capture materials, and methods of using same |
US10065185B2 (en) | 2007-07-13 | 2018-09-04 | Handylab, Inc. | Microfluidic cartridge |
US10071376B2 (en) | 2007-07-13 | 2018-09-11 | Handylab, Inc. | Integrated apparatus for performing nucleic acid extraction and diagnostic testing on multiple biological samples |
US8845980B2 (en) | 2007-09-10 | 2014-09-30 | Nec Corporation | Sample packing device |
JP2009145105A (ja) * | 2007-12-12 | 2009-07-02 | Konica Minolta Medical & Graphic Inc | マイクロチップ |
US8067176B2 (en) | 2008-01-25 | 2011-11-29 | Shimadzu Corporation | Microchemistry reaction method |
JP2009210327A (ja) * | 2008-03-03 | 2009-09-17 | Yokogawa Electric Corp | 化学反応用カートリッジ、混合物生成方法及び化学反応用カートリッジの制御装置 |
USD787087S1 (en) | 2008-07-14 | 2017-05-16 | Handylab, Inc. | Housing |
US9765389B2 (en) | 2011-04-15 | 2017-09-19 | Becton, Dickinson And Company | Scanning real-time microfluidic thermocycler and methods for synchronized thermocycling and scanning optical detection |
US10781482B2 (en) | 2011-04-15 | 2020-09-22 | Becton, Dickinson And Company | Scanning real-time microfluidic thermocycler and methods for synchronized thermocycling and scanning optical detection |
US11788127B2 (en) | 2011-04-15 | 2023-10-17 | Becton, Dickinson And Company | Scanning real-time microfluidic thermocycler and methods for synchronized thermocycling and scanning optical detection |
USD905269S1 (en) | 2011-09-30 | 2020-12-15 | Becton, Dickinson And Company | Single piece reagent holder |
US9480983B2 (en) | 2011-09-30 | 2016-11-01 | Becton, Dickinson And Company | Unitized reagent strip |
USD831843S1 (en) | 2011-09-30 | 2018-10-23 | Becton, Dickinson And Company | Single piece reagent holder |
US10076754B2 (en) | 2011-09-30 | 2018-09-18 | Becton, Dickinson And Company | Unitized reagent strip |
US11453906B2 (en) | 2011-11-04 | 2022-09-27 | Handylab, Inc. | Multiplexed diagnostic detection apparatus and methods |
US10822644B2 (en) | 2012-02-03 | 2020-11-03 | Becton, Dickinson And Company | External files for distribution of molecular diagnostic tests and determination of compatibility between tests |
US10220392B2 (en) | 2013-03-15 | 2019-03-05 | Becton, Dickinson And Company | Process tube and carrier tray |
US11433397B2 (en) | 2013-03-15 | 2022-09-06 | Becton, Dickinson And Company | Process tube and carrier tray |
US11865544B2 (en) | 2013-03-15 | 2024-01-09 | Becton, Dickinson And Company | Process tube and carrier tray |
JP2021081449A (ja) * | 2013-03-16 | 2021-05-27 | ドン ロバーツ レスリー | 自己完結型モジュラー分析カートリッジ及びプログラム可能試薬送達システム |
WO2024043160A1 (ja) * | 2022-08-23 | 2024-02-29 | 東洋製罐グループホールディングス株式会社 | 流体デバイス、流体デバイスの製造方法、及び検査システム |
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US7906318B2 (en) | 2011-03-15 |
CN1950504A (zh) | 2007-04-18 |
US20050250200A1 (en) | 2005-11-10 |
JPWO2005108571A1 (ja) | 2008-03-21 |
EP1746158A1 (en) | 2007-01-24 |
JP4784508B2 (ja) | 2011-10-05 |
CN100516212C (zh) | 2009-07-22 |
EP1746158A4 (en) | 2009-11-25 |
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