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公开(公告)号：US20100256007A1
公开(公告)日：2010-10-07
申请号：US12734235
申请日：2008-10-17
申请人： Mitsugu Usui , Chikako Hakii
发明人： Mitsugu Usui , Chikako Hakii
CPC分类号： G01N33/5308 , C12Q1/682 , C12Q2525/151
摘要： Provided are a method of forming a probe-polymer capable of readily and efficiently forming a polymer of a nucleic acid probe, a probe-polymer formed by the method, and a novel nucleic acid probe used in the method, and a detection method for a target analyte capable of sensitively and readily detecting the target analyte. The nucleic acid probe is formed by including three or more nucleic acid regions, in which each of the nucleic acid regions includes a first region and a second region complementary to the first region, the both regions being adjacent to each other. The polymer of the nucleic acid probe is formed by reacting the nucleic acid probe.
摘要翻译： 提供了形成能够容易且有效地形成核酸探针的聚合物的探针聚合物的方法，通过该方法形成的探针 - 聚合物和本方法中使用的新型核酸探针，以及检测方法 能够灵敏地容易地检测目标分析物的目标分析物。 核酸探针通过包括三个或更多个核酸区域形成，其中每个核酸区域包括第一区域和与第一区域互补的第二区域，两个区域彼此相邻。 核酸探针的聚合物通过核酸探针反应形成。
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2.发明申请Method for forming self-assembly substance using oligonucleotide synthesized by gene amplification reaction, self-assembly substance and method for detecting gene 失效使用通过基因扩增反应合成的寡核苷酸形成自组装物质的方法，自组装物质和检测基因的方法公开(公告)号：US20060286553A1
公开(公告)日：2006-12-21
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
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。
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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
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。
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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
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。
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5.发明授权Method for forming self-assembly substance using oligonucleotide synthesized by gene amplification reaction, self-assembly substance and method for detecting gene 失效使用通过基因扩增反应合成的寡核苷酸形成自组装物质的方法，自组装物质和检测基因的方法公开(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
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。
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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
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。
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7.发明授权Methods of constructing self-assembly of probes and method of detecting the same 失效构建探针自组装的方法及其检测方法公开(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
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。
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公开(公告)号：US07927804B2
公开(公告)日：2011-04-19
申请号：US12225062
申请日：2007-03-14
申请人： Chikako Hakii , Mitsugu Usui
发明人： Chikako Hakii , Mitsugu Usui
CPC分类号： C12Q1/682 , C12Q2565/501 , C12Q2563/179 , C12Q2537/125
摘要： Provided are a method of forming a signal probe-polymer which makes it possible to form a polymer efficiently and quantitatively, a signal probe-polymer formed by the method, oligonucleotide probes for use in the method, and a method of detecting target analyte having high sensitivity and excellent quantitative capability. The method of forming a signal probe-polymer comprises reacting a plurality of pairs of oligonucleotide probes with each other to form a polymer, a first probe of the pair of oligonucleotide probes comprising three nucleic acid regions of X, Y, and Z regions, located in the stated order from the 5′-terminal and a second probe comprising three nucleic acid regions of X′, Y′, and Z′ regions, located in the stated order from the 5′-terminal, wherein each region of the oligonucleotide probes has a length of from 13 to 15 bases.
摘要翻译： 提供了形成信号探针 - 聚合物的方法，其可以有效和定量地形成聚合物，通过该方法形成的信号探针 - 聚合物，用于该方法的寡核苷酸探针和检测具有高的靶分析物的方法 敏感性和优良的定量能力。 形成信号探针聚合物的方法包括使多对寡核苷酸探针彼此反应以形成聚合物，该对寡核苷酸探针的第一探针包含X，Y和Z区的三个核酸区，位于 按照5'末端的顺序，以及包含从5'末端起按照顺序的X'，Y'和Z'区域的三个核酸区域的第二探针，其中寡核苷酸探针的每个区域 长度为13至15个碱基。
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公开(公告)号：US20100248222A1
公开(公告)日：2010-09-30
申请号：US12225062
申请日：2007-03-14
申请人： Chikako Hakii , Mitsugu Usui
发明人： Chikako Hakii , Mitsugu Usui
CPC分类号： C12Q1/682 , C12Q2565/501 , C12Q2563/179 , C12Q2537/125
摘要： Provided are a method of forming a signal probe-polymer which makes it possible to form a polymer efficiently and quantitatively, a signal probe-polymer formed by the method, oligonucleotide probes for use in the method, and a method of detecting target analyte having high sensitivity and excellent quantitative capability. The method of forming a signal probe-polymer comprises reacting a plurality of pairs of oligonucleotide probes with each other to form a polymer, a first probe of the pair of oligonucleotide probes comprising three nucleic acid regions of X, Y, and Z regions, located in the stated order from the 5′-terminal and a second probe comprising three nucleic acid regions of X′, Y′, and Z′ regions, located in the stated order from the 5′-terminal, wherein each region of the oligonucleotide probes has a length of from 13 to 15 bases.
摘要翻译： 提供了形成信号探针 - 聚合物的方法，其可以有效和定量地形成聚合物，通过该方法形成的信号探针 - 聚合物，用于该方法的寡核苷酸探针和检测具有高的靶分析物的方法 敏感性和优良的定量能力。 形成信号探针聚合物的方法包括使多对寡核苷酸探针彼此反应以形成聚合物，该对寡核苷酸探针的第一探针包含X，Y和Z区的三个核酸区，位于 按照5'末端的顺序，以及包含从5'末端起按照顺序的X'，Y'和Z'区域的三个核酸区域的第二探针，其中寡核苷酸探针的每个区域 长度为13至15个碱基。
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10.发明授权Probe for constructing probe-polymer method of constructing probe-polymer and utilization thereof 失效探针聚合物构建探针 - 聚合物探针及其应用公开(公告)号：US07060814B2
公开(公告)日：2006-06-13
申请号：US09979999
申请日：2001-05-28
申请人： Mitsugu Usui , Chikako Hakii , Mari Mitsuka
发明人： Mitsugu Usui , Chikako Hakii , Mari Mitsuka
CPC分类号： C12N15/10 , C12Q1/6811 , C12Q1/6816 , C12Q1/682 , C12Q2527/101 , C12Q2525/185 , C12Q2525/313 , C12Q2563/179 , C12Q2537/143 , C12Q2525/161
摘要： The present invention provides a method for measuring a target gene under isothermal conditions without using enzyme. A pair of probes each having n (n≧3) base sequence regions complementary to each other are hybridized alternately to form a double-stranded probe-polymer. A base pair at branched sites of each complementary base sequence region is designed to be a G (guanine)-C (cytosine) bond, whereby a stable double-stranded probe-polymer is formed. One of complementary portions in one probe is constituted to have a base sequence complementary to a part of a target gene, whereby a target gene-probe-polymer complex is formed and the target gene is measured.
摘要翻译： 本发明提供了在不使用酶的条件下在等温条件下测定靶基因的方法。 一对具有互补互补的n（n> = 3）个碱基序列区的探针交替杂交以形成双链探针 - 聚合物。 每个互补碱基序列区的支链位点的碱基对设计为G（鸟嘌呤）-C（胞嘧啶）键，由此形成稳定的双链探针 - 聚合物。 一个探针中的互补部分之一构成为具有与靶基因的一部分互补的碱基序列，由此形成靶基因 - 探针 - 聚合物复合物，并测量靶基因。
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