公开(公告)号：US20100160179A1

公开(公告)日：2010-06-24

申请号：US12733165

申请日：2008-08-11

申请人： Mitsugu Usui ,  Chikako Hakii

发明人： Mitsugu Usui ,  Chikako Hakii

IPC分类号： C40B30/04 ,  C07H21/00 ,  C40B40/06

CPC分类号： C12Q1/682 ,  C12Q2537/143 ,  C12Q2525/313 ,  C12Q2537/162 ,  C12Q2537/125 ,  C12Q2525/161

摘要： Provided are a method of detecting a target substance whereby the detection sensitivity in the PALSAR method can be improved and multiple genes can be simultaneously detected, and a kit for detection. The method of detecting a target substance is a method of detecting a target substance by forming a signal probe polymer using one or more sets of paired dimer-forming probes for forming dimer probes or dimer probes, one or more kinds of crosslinking probes, and one or more kinds of assist probes, in which a dimer-forming probe includes a 5′-side region, a central region, and a 3′-side region, a crosslinking probe includes two regions, i.e., a 5′-side region and a 3′-side region, the 5′-side region of the dimer-forming probe is complementary to either 5′-side region of a crosslinking probe, the 3′-side region of the dimer-forming probe is complementary to either 3′-region of a crosslinking probe, and an assist probe includes a region complementary to the 5′-side region in one of paired dimer-forming probes, a region complementary to the 3′-side region in the other, and a target region capable of binding to the target substance.

摘要翻译： 提供了检测目标物质的方法，由此可以提高PALSAR方法中的检测灵敏度并同时检测多个基因，以及用于检测的试剂盒。 检测目标物质的方法是通过使用一组或多组用于形成二聚体探针或二聚体探针的配对二聚体形成探针，一种或多种交联探针和一种或多种交联探针形成信号探针聚合物来检测目标物质的方法 或多种辅助探针，其中二聚体形成探针包括5'-侧区，中心区和3'侧区域，交联探针包括两个区域，即5'侧区域和 3'-侧区域，二聚体形成探针的5'侧区域与交联探针的5'侧区域互补，二聚体形成探针的3'侧区域与3 交联探针的区域和辅助探针包括与成对的二聚体形成探针中的一个中的5'-侧区域互补的区域，与另一个的3'侧区域互补的区域和靶区域 能够结合目标物质。

公开(公告)号：US07632639B2

公开(公告)日：2009-12-15

申请号：US10546593

申请日：2004-02-20

申请人： Mitsugu Usui ,  Toshihiko Fujikawa

发明人： Mitsugu Usui ,  Toshihiko Fujikawa

IPC分类号： C12Q1/68 ,  C07H21/04

CPC分类号： C12Q1/682 ,  C12Q1/6827 ,  C12Q2565/113 ,  C12Q2565/501 ,  C12Q2561/125

摘要： There is provided a signal amplification method for detecting a mutated gene, which can increase the detection sensitivity of mutated genes on a DNA chip according to the PALSAR method, can establish efficient signal amplification and can establish simple detection by contriving design of oligonucleotide probes for use in the PALSAR method. The signal amplification method comprises a ligation reaction with a DNA ligase and a self-assembly reaction which forms a double-stranded self-assembly substance having a regular higher-order structure of oligonucleotides, wherein the detection sensitivity of the mutated gene on a DNA chip is improved.

摘要翻译： 提供了一种用于检测突变基因的信号扩增方法，其可以根据PALSAR方法增加DNA芯片上突变基因的检测灵敏度，可以建立有效的信号扩增，并且可以通过设计寡核苷酸探针来设计简单的检测来使用 在PALSAR方法中。 信号扩增方法包括与DNA连接酶的连接反应和形成具有规则的寡核苷酸高阶结构的双链自组装物质的自组装反应，其中突变基因在DNA芯片上的检测灵敏度 改进了

公开(公告)号：US07122310B2

公开(公告)日：2006-10-17

申请号：US10149187

申请日：2001-10-05

申请人： Mitsugu Usui ,  Mari Mitsuka ,  Chikako Hakii

发明人： Mitsugu Usui ,  Mari Mitsuka ,  Chikako Hakii

IPC分类号： C12Q1/68 ,  C12P19/34 ,  C07H21/04

CPC分类号： C12Q1/682 ,  C12N15/11 ,  C12Q1/6883 ,  C12Q2600/156 ,  G01N33/582

摘要： The present invention provides a novel method for forming a self-assembly substance of probes making it possible to shorten a reaction time required for formation of a self-assembly substance and also to increase regions of probes available for detection of a target gene by previously preparing a plurality of dimer-probes and increasing the types. This method comprises the steps of: providing plural groups, wherein each group includes a pair of dimer forming probes containing a pair of oligonucleotides, each oligonucleotide having three regions of a 3′ side region, a mid-region and a 5′ side region, in which the mid-regions of the oligonucleotides have base sequences complementary to each other, and the 3′ side regions and the 5′ side regions of the oligonucleotides have base sequences not complementary to each other; hybridizing a plurality of pairs of the dimer forming probes of the plural groups; and forming a double-stranded self-assembly substance by self-assembly of the oligonucleotides.

摘要翻译： 本发明提供了形成探针的自组装物质的新方法，使得可以缩短形成自组装物质所需的反应时间，并且还可以通过预先制备来增加可用于检测靶基因的探针的区域 多个二聚体探针并增加类型。 该方法包括以下步骤：提供多组，其中每组包括一对含有一对寡核苷酸的二聚体形成探针，每个寡核苷酸具有3'侧区域，中间区域和5'侧区域的三个区域， 其中寡核苷酸的中间区具有彼此互补的碱基序列，并且寡核苷酸的3'侧区和5'侧区具有彼此不互补的碱基序列; 杂交多对的二聚体形成探针对; 并通过寡核苷酸的自组装形成双链自组装物质。

公开(公告)号：US20060035235A1

公开(公告)日：2006-02-16

申请号：US10543683

申请日：2004-02-13

申请人： Mitsugu Usui ,  Toshihiko Fujikawa

发明人： Mitsugu Usui ,  Toshihiko Fujikawa

IPC分类号： C12Q1/68 ,  C12P19/34

CPC分类号： C12Q1/682 ,  C12Q2537/125

摘要： There is provided a signal amplification method for detecting an expressed gene, which allows detection with an inexpensive and simple operation in a short time without expensive enzymes and also allows detection depending on the length or expression amount of the original RNA without using the linear amplification method or the PCR method. In the signal amplification method, the detection sensitivity of the expressed gene on a DNA chip is improved by the use of a reverse transcription reaction and a self-assembly reaction forming a self-assembly substance by means of self-assembling of oligonucleotide probes.

摘要翻译： 提供了用于检测表达基因的信号放大方法，其允许在短时间内以便宜且简单的操作进行检测而没有昂贵的酶，并且还允许根据原始RNA的长度或表达量进行检测而不使用线性扩增方法 或PCR方法。 在信号扩增方法中，通过使用通过寡核苷酸探针的自组装形成自组装物质的逆转录反应和自组装反应，提高了DNA芯片上表达基因的检测灵敏度。

公开(公告)号：US07745124B2

公开(公告)日：2010-06-29

申请号：US11579063

申请日：2005-04-28

申请人： Chikako Hakii ,  Mitsugu Usui

发明人： Chikako Hakii ,  Mitsugu Usui

IPC分类号： C12Q1/68 ,  C07H21/02 ,  C07H21/04

CPC分类号： C12Q1/682 ,  B01L7/54 ,  C12Q1/6832 ,  Y10T436/143333 ,  C12Q2565/519 ,  C12Q2537/125 ,  C12Q2527/101

摘要： A hybridization method is provided in which an efficient hybridization reaction can be carried out. Further, there are provided, using this hybridization method, a method for detecting a target gene with high sensitivity and a signal amplifying method for markedly improving the detection sensitivity of the target gene. There is provided a hybridization method comprising the use of oligonucleotides in a reaction solution, the method comprising forming partially a reaction temperature region in the reaction solution and performing a hybridization reaction in the reaction temperature region.

摘要翻译： 提供可以进行有效的杂交反应的杂交方法。 此外，使用这种杂交方法，提供用于检测高灵敏度的靶基因的方法和用于显着提高靶基因的检测灵敏度的信号放大方法。 提供了包括在反应溶液中使用寡核苷酸的杂交方法，该方法包括部分地在反应溶液中形成反应温度区域并在反应温度区域进行杂交反应。

公开(公告)号：US20100151592A1

公开(公告)日：2010-06-17

申请号：US12733164

申请日：2008-08-11

申请人： Mitsugu Usui ,  Chikako Hakii

发明人： Mitsugu Usui ,  Chikako Hakii

IPC分类号： G01N33/53 ,  C07H21/00

CPC分类号： C12Q1/682 ,  C12Q2525/313

摘要： Provided are a method of detecting a target substance, by which the detection sensitivity in the PALSAR method can be improved and multiple genes can be simultaneously detected, and a kit for detection. The method is a method of detecting a target substance by forming a signal probe polymer with the use of multiple dimer probes or dimer-forming probes and an assist probe, in which each dimer probe is made up of two kinds of dimer-forming probes including a 5′-side region, a central region and a 3′-side region (the central regions of the two kinds of dimer-forming probes are complementary to each other and the 3′-side regions and the 5′-side regions thereof are not complementary to each other), the multiple dimer probes are constituted such that a dimer probe polymer is formed via a self-assembly reaction, and the assist probe is designed so as to have a structure having a region complementary to the 5′-side region of one dimer-forming probe in one dimer probe, a region complementary to the 3′-side region of the other dimer-forming probe, and a target region capable of binding to the target substance.

摘要翻译： 提供了检测目标物质的方法，通过该方法可以提高PALSAR方法中的检测灵敏度并同时检测多个基因，以及用于检测的试剂盒。 该方法是通过使用多个二聚体探针或二聚体形成探针和辅助探针形成信号探针聚合物来检测目标物质的方法，其中每个二聚体探针由两种二聚体形成探针组成，包括 5'侧区域，中央区域和3'侧区域（两种二聚体形成用探针的中心区域彼此互补，3'侧区域和5'侧区域 彼此不互补），构成多个二聚体探针，使得通过自组装反应形成二聚体探针聚合物，辅助探针被设计成具有与5'-末端互补的区域的结构， 在一个二聚体探针中的一个二聚体形成探针的侧面区域，与另一个二聚体形成探针的3'侧区域互补的区域和能够结合靶物质的靶区域。

公开(公告)号：US20090054809A1

公开(公告)日：2009-02-26

申请号：US11887832

申请日：2006-04-06

申请人： Takeharu Morishita ,  Mitsugu Usui

发明人： Takeharu Morishita ,  Mitsugu Usui

IPC分类号： A61B5/00

CPC分类号： G01N1/38 ,  C12Q1/24 ,  G01N2015/0088

摘要： It is intended to provide: a sampling device for quantitatively sampling a liquid sample having a high viscosity such as sputum; an auxiliary jig therefor; an agent for homogenizing sputum by which sputum can be quickly treated under mild conditions and a microbe contained in the sputum can be detected at an improved stability; a homogenization device; a homogenization method; a method of detecting a microbe by using the above method; a separation device for conveniently, quickly and efficiently collecting a microbe contained in sputum; a method of collecting a microbe; and a method of detecting a microbe by using the above method. A sampling device for a viscous sample includes: a tubular body; a gasket; a long rod-like plunger; a plate-shaped nozzle that has substantially the same diameter as that of a straight section of the tubular body and is adapted to close a distal end of the tubular body; and a slit formed in the plate-like nozzle, the slit having a predetermined shape adapted to serve as an aspiration and discharge port for aspirating and discharging the viscous sample.

摘要翻译： 旨在提供：用于定量采样具有高粘度的液体样品如痰液的取样装置; 一种辅助夹具; 可以在温和条件下快速治疗痰的痰匀浆剂，并且可以以更好的稳定性检测痰中含有的微生物; 均质装置; 均质化方法; 利用上述方法检测微生物的方法; 用于方便，快速和有效地收集痰中含有的微生物的分离装置; 收集微生物的方法; 以及使用上述方法检测微生物的方法。 用于粘性样品的采样装置包括：管状体; 垫圈 长棒状柱塞; 板状喷嘴，其直径与管状体的直线部分具有大致相同的直径，并且适于封闭管状体的远端; 以及形成在板状喷嘴中的狭缝，狭缝具有适于用作抽吸和排出粘性样品的抽吸和排出口的预定形状。

公开(公告)号：US07393636B2

公开(公告)日：2008-07-01

申请号：US10495065

申请日：2002-10-30

申请人： Mitsugu Usui ,  Mari Mitsuka ,  Chikako Hakii

发明人： Mitsugu Usui ,  Mari Mitsuka ,  Chikako Hakii

IPC分类号： C12Q1/68 ,  C12P19/34 ,  C07H21/04 ,  C07H21/02

CPC分类号： C12Q1/6844 ,  C12Q2525/155 ,  C12Q2525/313

摘要： There is provided a method for forming a self-assembly substance using oligonucleotides without using special instruments or complicated procedures, an self-assembly substance formed by the method for forming the self-assembly substance, and a method for detecting an amplified target gene at a low cost and in a simple way by making use of the method for forming the self-assembly substance. In the method for forming the self-assembly substance using a self-assembly reaction of oligonucleotides, the oligonucleotides comprise oligonucleotides synthesized by a gene amplification reaction. The oligonucleotides synthesized by the gene amplification reaction are detected by forming a self-assembly substance by the use of the method for forming the self-assembly substance of oligonucleotides, and by detecting the formed self-assembly substance.

摘要翻译： 提供了使用寡核苷酸而不使用特殊仪器或复杂程序形成自组装物质的方法，通过形成自组装物质的方法形成的自组装物质，以及用于检测扩增靶基因的方法 低成本，并且通过利用形成自组装物质的方法以简单的方式。 在使用寡核苷酸的自组装反应形成自组装物质的方法中，寡核苷酸包含通过基因扩增反应合成的寡核苷酸。 通过基因扩增反应合成的寡核苷酸通过使用形成寡核苷酸的自组装物质的方法形成自组装物质，并通过检测形成的自组装物质来检测。

公开(公告)号：US06261846B1

公开(公告)日：2001-07-17

申请号：US09292563

申请日：1999-04-15

申请人： Mitsugu Usui

发明人： Mitsugu Usui

IPC分类号： G01N3300

CPC分类号： C12N15/10 ,  C12Q1/6804 ,  C12Q1/682 ,  Y10T436/143333 ,  C12Q2565/125 ,  C12Q2525/313 ,  C12Q2525/107

摘要： A gene amplifying method for efficiently amplifying a gene without using enzyme or branched DNA. A plurality of pairs of probes each composed of three or more portions complementary to such portions in the other probe are used, and hybridized such that they cross in alternation to form a double-stranded polymer.

摘要翻译： 用于不使用酶或支链DNA有效扩增基因的基因扩增方法。 使用多个成对的探针，每个探针由在另一个探针中与这些部分互补的三个或更多个部分组成，并且杂交，使得它们交替交叉以形成双链聚合物。

公开(公告)号：US08450058B2

公开(公告)日：2013-05-28

申请号：US12733164

申请日：2008-08-11

申请人： Mitsugu Usui ,  Chikako Hakii

发明人： Mitsugu Usui ,  Chikako Hakii

IPC分类号： C12Q1/68 ,  C07H21/00 ,  C07H21/02 ,  C07H21/04

CPC分类号： C12Q1/682 ,  C12Q2525/313

摘要： Provided are a method of detecting a target substance, by which the detection sensitivity in the PALSAR method can be improved and multiple genes can be simultaneously detected, and a kit for detection. The method is a method of detecting a target substance by forming a signal probe polymer with the use of multiple dimer probes or dimer-forming probes and an assist probe, in which each dimer probe is made up of two kinds of dimer-forming probes including a 5′-side region, a central region and a 3′-side region (the central regions of the two kinds of dimer-forming probes are complementary to each other and the 3′-side regions and the 5′-side regions thereof are not complementary to each other), the multiple dimer probes are constituted such that a dimer probe polymer is formed via a self-assembly reaction, and the assist probe is designed so as to have a structure having a region complementary to the 5′-side region of one dimer-forming probe in one dimer probe, a region complementary to the 3′-side region of the other dimer-forming probe, and a target region capable of binding to the target substance.

摘要翻译： 提供了检测目标物质的方法，通过该方法可以提高PALSAR方法中的检测灵敏度并同时检测多个基因，以及用于检测的试剂盒。 该方法是通过使用多个二聚体探针或二聚体形成探针和辅助探针形成信号探针聚合物来检测目标物质的方法，其中每个二聚体探针由两种二聚体形成探针构成，包括 5'侧区域，中央区域和3'侧区域（两种二聚体形成用探针的中心区域彼此互补，3'侧区域和5'侧区域 彼此不互补），构成多个二聚体探针，使得通过自组装反应形成二聚体探针聚合物，辅助探针被设计成具有与5'-末端互补的区域的结构， 在一个二聚体探针中的一个二聚体形成探针的侧面区域，与另一个二聚体形成探针的3'侧区域互补的区域和能够结合靶物质的靶区域。