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公开(公告)号：US20190160466A1
公开(公告)日：2019-05-30
申请号：US16321766
申请日：2017-07-25
发明人： Yutaka YAMAGATA , Hiroyoshi AOKI , Kengo USUI , Yuki TANAKA , Norihiro MATSUYAMA , Kanji MITA
摘要： The present invention provides a tool that can analyze a target in a sample with simple operations and can be downsized, and an analysis method using the same. The analysis cell of the present invention includes: a main substrate: a sample inlet cover member; and a gas outlet cover member. The main substrate includes a flow path, an inlet for a sample, and a gas outlet, and the inlet and the gas outlet communicate with an outside. The inlet communicates with an upstream end portion of the flow path and the gas outlet communicates with a downstream end portion of the flow path. The flow path has a shape that expands from an upstream side toward a downstream side of the flow path. The sample inlet cover member is a liquid-tight member and can be fixed to the inlet when the sample inlet cover member is in use. The gas outlet cover member is a liquid-tight and gas-permeable member and can be fixed to the gas outlet when the gas outlet cover member is in use.
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2.发明授权公开(公告)号：US10760116B2
公开(公告)日：2020-09-01
申请号：US15755898
申请日：2016-08-26
IPC分类号： C12P19/34 , C12Q1/682 , C12Q1/68 , C12M1/00 , G01N21/64 , C12Q1/6818 , C12Q1/6851 , C12Q1/6876 , C12N15/09
摘要： The present invention provides a method for analyzing a template nucleic acid, a method for analyzing a target substance, an analysis kit for a template nucleic acid or a target substance, and an analyzer for a template nucleic acid or a target substance, which are excellent in accuracy. The method for analyzing a template nucleic acid of the present invention includes the steps of: fractionating a sample containing a template nucleic acid into a plurality of template nucleic acid fractions; amplifying a target sequence and its complementary sequence in the template nucleic acid with respect to each of the plurality of template nucleic acid fractions in the presence of a nucleic acid amplification reagent; detecting generation or quenching of a signal that shows an amplification of the target sequence or the complementary sequence with respect to each of the plurality of template nucleic acid fractions after the amplification step; and discriminating a template nucleic acid fraction in which the generation or quenching of a signal that shows the amplification has been detected among the plurality of template nucleic acid fractions as an amplified fraction in which the target sequence or the complementary sequence has been amplified, wherein the nucleic acid amplification reagent contains a primer set that amplifies the target sequence and the complementary sequence and a signal generating substance that generates or quenches a signal in response to the amplification, and the signal generating substance generates a signal in a state where it is bound sequence-dependently and quenches a signal in a state where it is not bound or quenches a signal in a state where it is bound sequence-dependently and generates a signal in a state where it is not bound, and generation and quenching of a signal are reversible.
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公开(公告)号：US20190032108A1
公开(公告)日：2019-01-31
申请号：US15755898
申请日：2016-08-26
申请人： RIKEN , KABUSHIKI KAISHA DNAFORM
IPC分类号： C12Q1/682 , G01N21/64 , C12Q1/6876
摘要： The present invention provides a method for analyzing a template nucleic acid, a method for analyzing a target substance, an analysis kit for a template nucleic acid or a target substance, and an analyzer for a template nucleic acid or a target substance, which are excellent in accuracy. The method for analyzing a template nucleic acid of the present invention includes the steps of: fractionating a sample containing a template nucleic acid into a plurality of template nucleic acid fractions; amplifying a target sequence and its complementary sequence in the template nucleic acid with respect to each of the plurality of template nucleic acid fractions in the presence of a nucleic acid amplification reagent; detecting generation or quenching of a signal that shows an amplification of the target sequence or the complementary sequence with respect to each of the plurality of template nucleic acid fractions after the amplification step; and discriminating a template nucleic acid fraction in which the generation or quenching of a signal that shows the amplification has been detected among the plurality of template nucleic acid fractions as an amplified fraction in which the target sequence or the complementary sequence has been amplified, wherein the nucleic acid amplification reagent contains a primer set that amplifies the target sequence and the complementary sequence and a signal generating substance that generates or quenches a signal in response to the amplification, and the signal generating substance generates a signal in a state where it is bound sequence-dependently and quenches a signal in a state where it is not bound or quenches a signal in a state where it is bound sequence-dependently and generates a signal in a state where it is not bound, and generation and quenching of a signal are reversible.
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公开(公告)号：US20170197216A1
公开(公告)日：2017-07-13
申请号：US15314373
申请日：2016-01-28
发明人： Koichiro TSUJIMARU
IPC分类号： B01L7/00
CPC分类号： B01L7/52 , B01L2300/0654 , B01L2300/0663 , B01L2300/168 , B01L2400/0478 , C12M1/00 , C12M1/34 , C12Q1/68 , G01N35/08 , G01N37/00
摘要： The present invention provides a tool that can analyze a target in a sample by a simple operation and can be downsized and an analysis method using the tool. By inserting an analysis chip into an analysis device, a reaction of a sample and a reagent is analyzed. The analysis chip includes a parallel flow channel in which plural reaction flow channels are connected in parallel, and the axial direction of the parallel flow channel is a direction of inserting the analysis chip into the analysis device. The analysis device includes a main body case being a housing, including an insertion opening into which the analysis chip is to be inserted; and a void; a heating unit that causes a sample to thermally react with a reagent, the heating unit being disposed in the void at at least one of inner surfaces so as to face a parallel surface of the parallel flow channel of the analysis chip; a light source unit that emits light to the analysis chip, the light source unit being disposed in the void at least above or below a reaction flow channel located at at least one end of the parallel flow channel of the analysis chip and extended along the axial direction of the parallel flow channel; and a plurality of detection units that detect the thermal reactions, each detection unit being disposed in the void at a downstream side end in the insertion direction of the analysis chip so as to correspond to each reaction flow channel of the analysis chip.
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5.发明申请公开(公告)号：US20190032127A1
公开(公告)日：2019-01-31
申请号：US16073210
申请日：2017-01-13
发明人： Yasuhiro MURAKAWA , Yujiro TAKEGAMI
IPC分类号： C12Q1/6869 , C12N15/113 , C12Q1/6806 , C12Q1/686
摘要： The present invention provides an analysis method by which a DNA element can be identified with higher sensitivity, and a transcription amount of the DNA element can be determined. The present invention is a method for decoding a base sequence of a nucleic acid corresponding to an end region of RNA, including: a preparing-decoding step of preparing, using a template RNA, at least one nucleic acid selected from the group consisting of: an RNA nucleic acid of a 5′-end region of the template RNA or a complementary DNA nucleic acid corresponding to the 5′-end region of the template RNA; an RNA nucleic acid of 3′-end region of the template RNA or a complementary DNA nucleic acid corresponding to the 3′-end region of the template RNA; an RNA nucleic acid of a partial region of an antisense strand or a sense strand for a full length of the template RNA; and combinations thereof or a nucleic acid having a sequence that has two or more linked sequences thereof, wherein the template RNA to be used in the preparing-decoding step is a nascent RNA. The DNA element can be identified and analyzed with higher accuracy and higher sensitivity by determining information on the decoded sequence of the nucleic acid and mapping the DNA element to the genome.
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公开(公告)号：US10066264B2
公开(公告)日：2018-09-04
申请号：US14414324
申请日：2013-08-29
发明人： Yoshihide Hayashizaki , Masayoshi Itoh , Takahiro Arakawa , Kengo Usui , Sotaro Uemura , Yasumasa Mitani
IPC分类号： C12Q1/68 , C12Q1/6876 , C12Q1/682 , C12Q1/6834 , C12Q1/6853
CPC分类号： C12Q1/6876 , C09B23/04 , C09B23/06 , C12Q1/682 , C12Q1/6834 , C12Q1/6853 , C12Q2600/158 , C12Q2525/301 , C12Q2565/1015 , C12Q2565/107 , C12Q2565/543
摘要： The present invention is to provide a method for analyzing a target nucleic acid, by which the target nucleic acid can be analyzed rapidly and easily. In order to achieve the above object, the present invention provides a method for analyzing a target nucleic acid in a sample, including the step of: analyzing the target nucleic acid in the sample by bringing the sample into contact with a label and with a primer or probe that can hybridize to the target nucleic acid. The primer or probe is immobilized on a solid phase. The label does not emit light when the primer or probe does not hybridize to the target nucleic acid, whereas the label emits light when the primer or probe has hybridized to the target nucleic acid. The analysis is carried out by detecting the light emitted from the label.
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公开(公告)号：US20180237834A1
公开(公告)日：2018-08-23
申请号：US15554604
申请日：2016-02-18
发明人： Koichiro TSUJIMARU
IPC分类号： C12Q1/6827 , C12Q1/6818 , G01N21/78
CPC分类号： C12Q1/6827 , C12M1/00 , C12N15/09 , C12Q1/6813 , C12Q1/6818 , G01N21/78 , C12Q2535/131 , C12Q2563/107 , C12Q2565/107 , C12Q2521/307 , C12Q2525/301
摘要： The present invention provides a method for accurately analyzing a template nucleic acid. The method is a method for analyzing a template nucleic acid including: an amplification step of amplifying a target sequence in the template nucleic acid and a complementary sequence to the target sequence; and a detection step of detecting association between a probe and an amplification product obtained in the amplification step or dissociation of an assembly between the probe and the amplification product with a temperature change. A primer set is used in the amplification step, and the primer set is a combination of a first primer for a polymerase that performs priming at the 3′ end of a primer to synthesize the target sequence and a second primer for the polymerase to synthesize the complementary strand. Between the first primer and the second primer, a primer that initiates synthesis of a sequence to be hybridized with the probe is a turnback primer including a primer region and a probe region being directly or indirectly linked to the 5′ end of the primer region, and the other primer is any primer having an ability of causing the polymerase to perform priming at the 3′ end. In the amplification step, the turnback primer extends the target sequence or the complementary sequence from the primer region. In the detection step, the turnback primer also serves as the probe, and association between the probe region and the target sequence or the complementary sequence, occurred by turning the probe region of the turnback primer in a molecule of the amplification product back toward the extended target sequence or the extended complementary sequence, is detected
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公开(公告)号：US11312991B2
公开(公告)日：2022-04-26
申请号：US16073210
申请日：2017-01-13
发明人： Yasuhiro Murakawa , Yujiro Takegami
IPC分类号： C12Q1/68 , C12Q1/6869 , C12N15/10 , C12N15/113 , C12Q1/6806 , C12Q1/686 , C07H21/04 , C07H21/00
摘要： The present invention provides an analysis method by which a DNA element can be identified with higher sensitivity, and a transcription amount of the DNA element can be determined. The present invention is a method for decoding a base sequence of a nucleic acid corresponding to an end region of RNA, including: a preparing-decoding step of preparing, using a template RNA, at least one nucleic acid selected from the group consisting of: an RNA nucleic acid of a 5′-end region of the template RNA or a complementary DNA nucleic acid corresponding to the 5′-end region of the template RNA; an RNA nucleic acid of 3′-end region of the template RNA or a complementary DNA nucleic acid corresponding to the 3′-end region of the template RNA; an RNA nucleic acid of a partial region of an antisense strand or a sense strand for a full length of the template RNA; and combinations thereof or a nucleic acid having a sequence that has two or more linked sequences thereof, wherein the template RNA to be used in the preparing-decoding step is a nascent RNA. The DNA element can be identified and analyzed with higher accuracy and higher sensitivity by determining information on the decoded sequence of the nucleic acid and mapping the DNA element to the genome.
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公开(公告)号：US10428371B2
公开(公告)日：2019-10-01
申请号：US15300183
申请日：2015-03-27
IPC分类号： C12Q1/68 , C12Q1/6818 , C12Q1/6816 , C09B23/04 , C09B23/06
摘要： The present invention is intended to provide a novel fluorescent labeled single-stranded nucleic acid, by which the background of an exciton oligomer can be further reduced and the novel use thereof. The present invention relates to a labeled single-stranded nucleic acid having at least two fluorescent atomic group pairs that exhibit an exciton effect. The labeled single-stranded nucleic acid is characterized in that the emission peak wavelength of one of the fluorescent atomic group pairs (fluorescent atomic group pair A) is shorter than the excitation peak wavelength of the other fluorescent atomic group pair (fluorescent atomic group pair B), and the fluorescent atomic group pairs A and B have a Förster resonance energy transfer (FRET) effect. This fluorescent labeled single-stranded nucleic acid is usable as a primer for amplifying a target nucleic acid or a probe to be hybridized with a target nucleic acid.
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公开(公告)号：US20180371518A1
公开(公告)日：2018-12-27
申请号：US15747044
申请日：2016-04-27
发明人： Koichiro TSUJIMARU
摘要： The present invention provides a miniaturizable tool that can analyze a target in a sample by a simple operation and an analysis method using the tool. An analysis chip is a syringe-type chip including: a syringe main body having an openable and closable tip; and a piston to be inserted into the syringe main body, the piston being a hollow body having a translucent closed-end at a side to be inserted into the syringe main body and an open-end at the other side. Analysis device includes a main body case that is a housing, the housing including: an insertion opening into which the syringe-type chip is to be inserted; and a void in communication with the insertion opening, wherein an opposite end of the insertion opening in an axial direction is a bottom; a heating unit that heats the syringe-type chip; a light source unit that emits light to the syringe-type chip; and a detection unit that is a columnar body that can be inserted into the hollow body of the piston and disposed at the bottom of the housing in the axial direction. In use, the syringe-type chip is inserted into the analysis device from a side of the piston and the detection unit of the analysis device is inserted into the piston of the syringe-type chip.
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