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公开(公告)号：US20090137406A1
公开(公告)日：2009-05-28
申请号：US12227904
申请日：2007-06-01
申请人： Kenji Kinoshita , Kanehisa Yokoyama , Kentaro Fujimoto , Toru Yakabe , Shin Saito , Kazuhiko Fujiwara
发明人： Kenji Kinoshita , Kanehisa Yokoyama , Kentaro Fujimoto , Toru Yakabe , Shin Saito , Kazuhiko Fujiwara
CPC分类号： C12Q1/6837
摘要： The present invention provides an RNA detection method detecting, from a reaction system containing a target sample, a target RNA chain originated from the target sample, using a surface having on the surface thereof a polymer substance which contains a first unit having a group derived from a phosphate ester composing the hydrophilic portion of a phospholipid and a second unit having a carboxylic acid derivative group composed of an electron-attractive substitutional group bound to a carbonyl group, while being provided with at least one reaction space, the reaction space having an immobilized nucleic acid primer immobilized therein.
摘要翻译： 本发明提供一种RNA检测方法，其从含有目标样品的反应体系中检测源自目标样品的靶RNA链，其表面上具有含有第一单元的表面，该第一单元含有来自 构成磷脂的亲水部分的磷酸酯和具有由与羰基结合的吸电子取代基组成的羧酸衍生物基团的第二单元，同时具有至少一个反应空间，所述反应空间具有固定化的 固定在其中的核酸引物。
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公开(公告)号：US08088580B2
公开(公告)日：2012-01-03
申请号：US12227904
申请日：2007-06-01
申请人： Kenji Kinoshita , Kanehisa Yokoyama , Kentaro Fujimoto , Toru Yakabe , Shin Saito , Kazuhiko Fujiwara
发明人： Kenji Kinoshita , Kanehisa Yokoyama , Kentaro Fujimoto , Toru Yakabe , Shin Saito , Kazuhiko Fujiwara
CPC分类号： C12Q1/6837
摘要： The present invention provides an RNA detection method detecting, from a reaction system containing a target sample, a target RNA chain originated from the target sample, using a surface having on the surface thereof a polymer substance which contains a first unit having a group derived from a phosphate ester composing the hydrophilic portion of a phospholipid and a second unit having a carboxylic acid derivative group composed of an electron-attractive substitutional group bound to a carbonyl group, while being provided with at least one reaction space, the reaction space having an immobilized nucleic acid primer immobilized therein.
摘要翻译： 本发明提供一种RNA检测方法，其从含有目标样品的反应体系中检测源自目标样品的靶RNA链，其表面上具有含有第一单元的表面，该第一单元含有来自 构成磷脂的亲水部分的磷酸酯和具有由与羰基结合的吸电子取代基组成的羧酸衍生物基团的第二单元，同时具有至少一个反应空间，所述反应空间具有固定化的 固定在其中的核酸引物。
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3.发明申请Method of synthesizing cdna and method of synthesizing rna chains, and nucleotide-immobilized carrier 有权合成cdna的方法和合成rna链的方法以及核苷酸固定载体公开(公告)号：US20100016566A1
公开(公告)日：2010-01-21
申请号：US11990641
申请日：2006-08-09
CPC分类号： C12P19/34 , C12N15/1096 , C12Q1/686 , C12Q2521/107 , C12Q2565/537
摘要： A method of synthesizing a cDNA chain using an insoluble carrier having on the surface thereof a polymer substance containing a first unit having a group derived from a phosphoric ester composing the hydrophilic portion of phospholipid, and a second unit having a group derived from carboxylic acid having an electron-attractive substituent bound to a carbonyl group, which includes immobilizing a polynucleotide for DNA elongation; bringing a solution containing an RNA fragment, nucleotide monomers, and a reverse transcriptase or an enzyme having polymerase activity into contact with the surface of the insoluble carrier; and allowing the polynucleotide for DNA elongation immobilized on the surface of the carrier to elongate using the RNA fragment contained in the solution as a template, to thereby form a single-strand cDNA.
摘要翻译： 一种使用不溶性载体合成cDNA链的方法，所述不溶性载体在其表面上具有含有源自构成磷脂的亲水部分的磷酸酯的基团的第一单元的聚合物物质和具有衍生自羧酸的基团的第二单元， 与羰基结合的电子吸引取代基，其包括固定多核苷酸用于DNA延伸; 将含有RNA片段，核苷酸单体和逆转录酶或具有聚合酶活性的酶的溶液与不溶性载体的表面接触; 并且使用溶液中包含的RNA片段作为模板，使固定在载体表面上的DNA延伸的多核苷酸伸长，从而形成单链cDNA。
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公开(公告)号：US07968290B2
公开(公告)日：2011-06-28
申请号：US11920559
申请日：2006-05-16
CPC分类号： C12N15/1096 , C12Q1/6834 , C12Q2565/537 , C12Q2527/107
摘要： A method of detecting a gene including immobilizing a primer for DNA elongation onto an insoluble carrier having on the surface thereof a polymer substance containing a first unit having a phosphorylcholine group and a second unit having a carboxylic acid-derived group having an electron-attractive substituent bound to a carbonyl group; annealing the template DNA fragments or RNA fragments with the primer for DNA elongation, so as to elongate the DNA primer while incorporating therein an enzyme, thereby allowing coloration of a chromogenic reagent by its enzymatic action; and judging whether the DNA fragments or RNA fragments of the gene presents or not, based on the degree of coloration.
摘要翻译： 一种检测基因的方法，包括将DNA延伸引物固定在具有表面的不溶性载体上，所述不溶性载体具有含有具有磷酸胆碱基团的第一单元的聚合物物质和具有吸电子取代基的羧酸衍生基团的第二单元 与羰基结合; 用引物退火模板DNA片段或RNA片段以进行DNA延伸，以便在引入酶的同时延长DNA引物，从而通过其酶作用使着色试剂着色; 并基于着色程度来判断该基因的DNA片段或RNA片段是否存在。
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5.发明授权Method of synthesizing cDNA and method of synthesizing RNA chains, and nucleotide-immobilized carrier 有权合成cDNA的方法和合成RNA链的方法以及核苷酸固定载体公开(公告)号：US08822670B2
公开(公告)日：2014-09-02
申请号：US11990641
申请日：2006-08-09
IPC分类号： C07H21/00
CPC分类号： C12P19/34 , C12N15/1096 , C12Q1/686 , C12Q2521/107 , C12Q2565/537
摘要： A method of synthesizing a cDNA chain using an insoluble carrier having on the surface thereof a polymer substance containing a first unit having a group derived from a phosphoric ester composing the hydrophilic portion of phospholipid, and a second unit having a group derived from carboxylic acid having an electron-attractive substituent bound to a carbonyl group, which includes immobilizing a polynucleotide for DNA elongation; bringing a solution containing an RNA fragment, nucleotide monomers, and a reverse transcriptase or an enzyme having polymerase activity into contact with the surface of the insoluble carrier; and allowing the polynucleotide for DNA elongation immobilized on the surface of the carrier to elongate using the RNA fragment contained in the solution as a template, to thereby form a single-strand cDNA.
摘要翻译： 一种使用不溶性载体合成cDNA链的方法，所述不溶性载体在其表面上具有含有源自构成磷脂的亲水部分的磷酸酯的基团的第一单元的聚合物物质和具有衍生自羧酸的基团的第二单元， 与羰基结合的电子吸引取代基，其包括固定多核苷酸用于DNA延伸; 将含有RNA片段，核苷酸单体和逆转录酶或具有聚合酶活性的酶的溶液与不溶性载体的表面接触; 并且使用溶液中包含的RNA片段作为模板，使固定在载体表面上的DNA延伸的多核苷酸伸长，从而形成单链cDNA。
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公开(公告)号：US20090042190A1
公开(公告)日：2009-02-12
申请号：US11920559
申请日：2006-05-16
IPC分类号： C12Q1/68
CPC分类号： C12N15/1096 , C12Q1/6834 , C12Q2565/537 , C12Q2527/107
摘要： A method of detecting a gene including immobilizing a primer for DNA elongation onto an insoluble carrier having on the surface thereof a polymer substance containing a first unit having a phosphorylcholine group and a second unit having a carboxylic acid-derived group having an electron-attractive substituent bound to a carbonyl group; annealing the template DNA fragments or RNA fragments with the primer for DNA elongation, so as to elongate the DNA primer while incorporating therein an enzyme, thereby allowing coloration of a chromogenic reagent by its enzymatic action; and judging whether the DNA fragments or RNA fragments of the gene presents or not, based on the degree of coloration.
摘要翻译： 一种检测基因的方法，包括将DNA延伸引物固定在具有表面的不溶性载体上，所述不溶性载体具有含有具有磷酸胆碱基团的第一单元的聚合物物质和具有吸电子取代基的羧酸衍生基团的第二单元 与羰基结合; 用引物退火模板DNA片段或RNA片段以进行DNA延伸，以便在引入酶的同时延长DNA引物，从而通过其酶作用使着色试剂着色; 并基于着色程度来判断该基因的DNA片段或RNA片段是否存在。
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公开(公告)号：US08435742B2
公开(公告)日：2013-05-07
申请号：US12739978
申请日：2008-11-04
CPC分类号： C12Q1/6809 , C12Q2565/537 , C12Q2525/143 , C12Q2521/107
摘要： [Object] It is to provide a method and a kit capable of detecting or quantifying a target RNA simply and rapidly from trace amounts of RNA in a sample, in a case such as when one or more kinds of pathogenic microorganisms are to be detected or quantified.[Solving Means] The method comprises the steps of 1) synthesizing cDNA from a sample containing the target RNA using a liquid-phase primer having a promoter sequence and a reverse transcriptase to obtain a cDNA-RNA complex, 2) degrading the RNA of the complex, 3) synthesizing a double-stranded DNA via the cDNA obtained in the step 2) and the solid-phase primer, 4) synthesizing RNA from the double-stranded DNA, 5) synthesizing cDNA via the RNA obtained in the step 4) and the solid-phase primer to obtain a cDNA-RNA complex, 6) degrading the RNA of the complex obtained in step 5), 7) synthesizing a double-stranded DNA via the cDNA obtained in the step 6) and the liquid-phase primer, and 8) quantifying the double-stranded DNAs obtained in the steps 3) and 7). These steps can be performed in a single reaction solution.
摘要翻译： 本发明提供一种方法和试剂盒，其能够在样品中的痕量RNA中简单快速地检测或定量靶RNA，例如当检测到一种或多种病原微生物或 量化 [解决方法]该方法包括以下步骤：1）使用具有启动子序列和逆转录酶的液相引物合成来自含有靶RNA的样品的cDNA，得到cDNA-RNA复合物，2）降解 复合物，3）通过步骤2）和固相引物获得的cDNA合成双链DNA，4）从双链DNA合成RNA，5）通过步骤4）得到的RNA合成cDNA， 和固相引物以获得cDNA-RNA复合物，6）降解步骤5）中获得的复合物的RNA，7）通过步骤6）中得到的cDNA合成双链DNA和液相 引物，和8）定量在步骤3）和7）中获得的双链DNA。 这些步骤可以在单个反应溶液中进行。
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公开(公告)号：US20110053150A1
公开(公告)日：2011-03-03
申请号：US12739978
申请日：2008-11-04
IPC分类号： C12Q1/68
CPC分类号： C12Q1/6809 , C12Q2565/537 , C12Q2525/143 , C12Q2521/107
摘要： [Object] It is to provide a method and a kit capable of detecting or quantifying a target RNA simply and rapidly from trace amounts of RNA in a sample, in a case such as when one or more kinds of pathogenic microorganisms are to be detected or quantified.[Solving Means] The method comprises the steps of 1) synthesizing cDNA from a sample containing the target RNA using a liquid-phase primer having a promoter sequence and a reverse transcriptase to obtain a cDNA-RNA complex, 2) degrading the RNA of the complex, 3) synthesizing a double-stranded DNA via the cDNA obtained in the step 2) and the solid-phase primer, 4) synthesizing RNA from the double-stranded DNA, 5) synthesizing cDNA via the RNA obtained in the step 4) and the solid-phase primer to obtain a cDNA-RNA complex, 6) degrading the RNA of the complex obtained in step 5), 7) synthesizing a double-stranded DNA via the cDNA obtained in the step 6) and the liquid-phase primer, and 8) quantifying the double-stranded DNAs obtained in the steps 3) and 7). These steps can be performed in a single reaction solution.
摘要翻译： 本发明提供一种方法和试剂盒，其能够在样品中的痕量RNA中简单快速地检测或定量靶RNA，例如当检测到一种或多种病原微生物或 量化 [解决方法]该方法包括以下步骤：1）使用具有启动子序列和逆转录酶的液相引物合成来自含有靶RNA的样品的cDNA，得到cDNA-RNA复合物，2）降解 复合物，3）通过步骤2）和固相引物获得的cDNA合成双链DNA，4）从双链DNA合成RNA，5）通过步骤4）得到的RNA合成cDNA， 和固相引物以获得cDNA-RNA复合物，6）降解步骤5）中获得的复合物的RNA，7）通过步骤6）中得到的cDNA合成双链DNA和液相 引物，和8）定量在步骤3）和7）中获得的双链DNA。 这些步骤可以在单个反应溶液中进行。
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公开(公告)号：US3935193A
公开(公告)日：1976-01-27
申请号：US102556
申请日：1970-12-29
申请人： Jinnosuke Abe , Tetsuo Watanabe , Teruo Take , Kentaro Fujimoto , Tadashiro Fujii , Kazuyoshi Nishiie
发明人： Jinnosuke Abe , Tetsuo Watanabe , Teruo Take , Kentaro Fujimoto , Tadashiro Fujii , Kazuyoshi Nishiie
IPC分类号： C07D499/00 , C07D499/04 , C07D499/44 , C07D99/16
CPC分类号： C07D499/00 , Y02P20/55
摘要： Novel diacyl penicillins of the formula ##EQU1## wherein R is alkyl, cycloalkyl, aryl, aralkyl, phenoxyalkyl, heterocyclic carboxyl excepting isoxazole carboxyl derivatives, or ##SPC1##In which benzene ring A may optionally be substituted andB represents a protective group for the amino group, and X is a protective group for the carboxyl group, are produced by reacting a benzyl penicillin ester of the formula ##EQU2## wherein X is as defined above, with a chlorinating agent in the presence of tertiary organic base to obtain an imide chloride group-incorporated compound of the formula ##EQU3## and then reacting the compound of the last-named formula with carboxylate of the formulaR-COOM (IV)wherein M is a metal atom, and R is as defined above.
摘要翻译： 式O CH2-C的新型二酰基青霉素O CH2-C ANGLE S ANGLE CH3 N-CH-CHC ANGLE |||CH3 RC ANGLE ||| O||| O = C-N --- CH-COOX（I）其中R 或烷基，环烷基，芳基，芳烷基，苯氧基烷基，除异恶唑羧基衍生物以外的杂环羧基，或其中任选取代的苯甲烯环，B表示氨基的保护基，X为羧基的保护基， 通过使式S ANGLE CH3 CH2-CO-NH-CH-CHC ANGLE |||CH3 O = C-N --- CH-COOX（II）的苄基青霉素酯反应制备，其中X如上定义， 在叔有机碱的存在下与氯化剂反应，得到式I化合物的酰亚胺氯化物基团化合物。ANGLE CH3 CH2-C = N-CH-CHC ANGLE ||| CH3 ClO = C-N --- CH-COOX，（III），然后使最后命名的化合物与式R-COOM（Ⅳ）的羧酸酯反应，其中M是金属原子，R如上定义。
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