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公开(公告)号：US20070238125A1
公开(公告)日：2007-10-11
申请号：US11783124
申请日：2007-04-06
CPC分类号： C12P19/34
摘要： The present invention relates to a probe synthesis method and a probe synthesis kit for nucleic acid detection, which are intended for the fluorophore labeling and detection of a target gene. In this method, a probe unit for analyte nucleic acid recognition having a base sequence specific to an analyte nucleic acid and plural labeled probe units each having a fluorophore labeled internal base are hybridized to oligonucleotides complementary thereto, and the adjacent probe units are bonded by ligation. As a result, an analytical probe with increased fluorescence intensity is synthesized easily and inexpensively, while the number of fluorophore labels is controlled.
摘要翻译： 本发明涉及用于靶基因的荧光标记和检测的用于核酸检测的探针合成方法和探针合成试剂盒。 在该方法中，具有分析物核酸特异性的碱基序列的分析物核酸识别用探针单元和各自具有荧光团标记的内部碱基的多个标记的探针单元与互补的寡核苷酸杂交，并且通过连接键合相邻的探针单元 。 结果，容易且廉价地合成了增加荧光强度的分析探针，同时控制了荧光标记的数量。
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公开(公告)号：US20060040261A1
公开(公告)日：2006-02-23
申请号：US11132286
申请日：2005-05-19
申请人： Maiko Tanabe , Chihiro Uematsu
发明人： Maiko Tanabe , Chihiro Uematsu
CPC分类号： C12Q1/707
摘要： The present application relates to primers for isothermal amplification of HCV each include at least eighteen consecutive bases corresponding to a 3′ end region of one selected from base sequences of SEQ ID NOs: 1-10, 21 and 22. The primers are specific to HCV subtypes 1a, 1b, 2a, 2b and 3a, respectively and enable genotyping of HCV by isothermal amplification.
摘要翻译： 本申请涉及用于HCV等温扩增的引物各自包括至少十八个连续的碱基，其对应于选自SEQ ID NO：1-10,21和22的碱基序列的3'端区域。引物对HCV特异 亚型1a，1b，2a，2b和3a，使得能够通过等温扩增进行HCV的基因分型。
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公开(公告)号：US06242193B1
公开(公告)日：2001-06-05
申请号：US09567270
申请日：2000-05-09
IPC分类号： C12Q168
CPC分类号： G01N21/6458 , C12Q1/6869 , G01N21/648 , G01N21/6486 , C12Q2537/149 , C12Q2523/319
摘要： A single molecule of single-stranded sample DNA (7) having a bead (5) at one end and a magnetic bead (6) at the other end is extended and fixed in the field of view of a fluorescent microscope by using a magnetic force (11) and a laser trap (3), and a primer (8) is bonded thereto, followed by elongation reaction (10) using polymerase. Only a single chemically modified nucleotide (9) labeled with at least one fluorophore which varies depending on the kind of the base is incorporated. Only the single fluorophore incorporated is measured as a fluorescence-microscopic image by evanescent irradiation (13) with exciting laser beams, and the kind of the base is determined from the kind of the fluorophore. The fluorophore labeling the nucleotide incorporated is released by evanescent irradiation (13) with ultraviolet laser beams (2), and the next nucleotide is incorporated. DNA sequencing is carried out by repeating the above procedure. The base sequence determination can be carried out by using the single DNA molecule, so that a DNA base sequence of hundreds kilos or more bases can be efficiently determined.
摘要翻译： 在一端具有珠（5）和另一端的磁珠（6）的单分子单链样品DNA（7）在荧光显微镜的视场中通过使用磁力 （11）和激光捕获器（3），并将引物（8）与其结合，然后使用聚合酶进行伸长反应（10）。 只有一个化学修饰的核苷酸（9）被至少一个根据碱的种类而变化的荧光团标记。 通过用激发激光束的ev逝照射（13）测量所掺入的单个荧光团作为荧光显微镜图像，并且根据荧光团的种类确定碱基的种类。 标记所结合核苷酸的荧光团通过用紫外激光束（2）的ev逝照射（13）释放，并且并入下一个核苷酸。 通过重复上述步骤进行DNA测序。 碱基序列的测定可以通过使用单个DNA分子进行，从而可以有效地确定数百个碱基的DNA碱基序列。
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公开(公告)号：US06183970B2
公开(公告)日：2001-02-06
申请号：US09383198
申请日：1999-08-26
申请人： Kazunori Okano , Hideki Kambara , Chihiro Uematsu , Hiroko Matsunaga , Takashi Irie , Tomoharu Kajiyama , Kenji Yasuda
发明人： Kazunori Okano , Hideki Kambara , Chihiro Uematsu , Hiroko Matsunaga , Takashi Irie , Tomoharu Kajiyama , Kenji Yasuda
IPC分类号： C12Q168
CPC分类号： B82Y30/00 , B01J19/0046 , B01J2219/00317 , B01J2219/00576 , B01J2219/00596 , B01J2219/00608 , B01J2219/0061 , B01J2219/00612 , B01J2219/00617 , B01J2219/00619 , B01J2219/00621 , B01J2219/00626 , B01J2219/00637 , B01J2219/00641 , B01J2219/00644 , B01J2219/00653 , B01J2219/00659 , B01J2219/00677 , B01J2219/00702 , B01J2219/00704 , B01J2219/00713 , B01J2219/00722 , B01L3/5085 , B01L3/5088 , B01L2300/0819 , B01L2400/0421 , C12Q1/6837 , C40B40/06 , C12Q2565/607 , C12Q2561/119
摘要： There are beforehand prepared a monomer having a reaction residue and a polynucleotide probe set comprising plural kinds of polynucleotide probes having a residue bonded to the reaction residue. The monomer is mixed with each kind of polynucleotide probes comprising any plural probes selected from the polynucleotide probe set. Each kind of the resultant mixtures is added to each of different small holes to make the mixture into gel matrix. Thus, a polynucleotide probe chip is produced. Sample DNA is forcibly migrated in the gels by electrophoresis. Laser light is projected onto the side face of the chip. The fluorescence emitted from the whole surface of the chip is collectively detected with a high-sensitive two-dimensional detector. Thus, the polynucleotide probe chip, holding various kinds of DNA probes, for detecting DNA can be provided. This chip has high hybridization-efficiency and makes high-sensitivity and high-speed DNA detection possible.
摘要翻译： 预先制备具有反应残基的单体和含有残基与反应残基连接的多种多核苷酸探针的多核苷酸探针组。 将单体与包含选自多核苷酸探针组的任何多个探针的各种多核苷酸探针混合。 将各种所得混合物加入每个不同的小孔中以使混合物成为凝胶基质。 因此，产生多核苷酸探针芯片。 样品DNA通过电泳在凝胶中强制迁移。 激光投射到芯片的侧面。 从芯片的整个表面发射的荧光由高灵敏度二维检测器共同检测。 因此，可以提供用于检测DNA的保持各种DNA探针的多核苷酸探针芯片。 该芯片具有高杂交效率，可实现高灵敏度和高速DNA检测。
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公开(公告)号：US5948615A
公开(公告)日：1999-09-07
申请号：US834385
申请日：1997-04-16
申请人： Chihiro Uematsu , Hideki Kambara
发明人： Chihiro Uematsu , Hideki Kambara
CPC分类号： C12N15/1096 , C12Q1/683 , C12Q1/6855
摘要： The present invention comprises a method for analysis of a nucleic acid which comprises:(1) a step of digesting a double-stranded DNA sample with a plurality of restriction enzymes to obtain double-stranded DNA fragments;(2) a step of ligating a plurality of oligonucleotides to the double-stranded DNA fragments respectively at the both ends thereof;(3) a step of dispensing a solution containing the double-stranded DNA fragments into a plurality of tubes;(4) a step of adding DNA primers comprising combinations of DNA primers selected from each set of a plurality of DNA primer sets comprising a plurality of labeled primers having a base sequence complementary to the base sequence of oligonucleotide and a part or all of the base sequence contiguous to the base sequence complementary to the base sequence of the oligonucleotide and recognized by the restriction enzymes and a selective base sequence of 1 to 4 bases at the 3'-end thereof, to the respective tubes corresponding to the combinations and performing a complementary strand synthesis reaction of the region of the double-stranded DNA fragments between the base sequences recognized by the two restriction enzymes; and,(5) a step of subjecting the products obtained by the complementary strand synthesis reaction to electrophoresis to produce a large number of DNA fragments from the long double stranded DNA sample digested with restriction enzymes and obtain fingerprinting patterns therefrom which enables to inspect the long double-stranded DNA sample.
摘要翻译： 本发明包括核酸分析方法，其包括：（1）用多种限制酶消化双链DNA样品以获得双链DNA片段的步骤; （2）在其两端分别将多个寡核苷酸与双链DNA片段连接的步骤; （3）将含有双链DNA片段的溶液分配到多个管中的步骤; （4）添加DNA引物的步骤，其包含选自每组多个DNA引物组的DNA引物的组合，所述多个DNA引物组包含多个具有与寡核苷酸的碱基序列互补的碱基序列的标记引物和碱基的一部分或全部 与寡核苷酸的碱基序列互补的碱基序列连续的序列，并且被限制性内切酶和在其3'末端具有1至4个碱基的选择性碱基序列连接到对应于组合的各个管，并进行互补 在两个限制酶识别的碱基序列之间的双链DNA片段的区域的链合成反应; 和（5）使由互补链合成反应得到的产物进行电泳，从用限制酶消化的长双链DNA样品中产生大量DNA片段的步骤，并从中获得指纹图案，其能够检查长 双链DNA样品。
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6.发明授权公开(公告)号：US09221055B2
公开(公告)日：2015-12-29
申请号：US13883830
申请日：2011-10-27
CPC分类号： B01L7/52 , B01L3/50851 , B01L3/50855 , B01L7/5255 , B01L2200/025 , B01L2300/0803 , B01L2300/0822 , G01N21/6456 , G01N21/6486 , G01N35/028 , G01N2035/00366 , G01N2035/0429 , G01N2035/0441 , G01N2035/0465
摘要： Nucleic acid analysis apparatus includes a plurality of temperature adjustment apparatuses, a rotating mechanism, a delivery base and an ejection base, a delivery drive mechanism, an ejection drive mechanism, and a detection apparatus. The rotating mechanism can include a rotating shaft and a plurality of pressing portions that rotate around the rotating shaft. The reaction plate assembly can move over the temperature adjustment apparatuses along the circumferential direction in a state of being pressed onto the temperature adjustment apparatuses by the pressing portions. The delivery drive mechanism can cause the reaction plate assembly to be moved radially inward and delivered between the pressing portions and temperature adjustment apparatuses. The ejection drive mechanism can cause the reaction plate assembly to be moved radially outward and ejected from between the pressing portions and temperature adjustment apparatuses onto the ejection base. Reaction plates, reaction plate assemblies, and nucleic acid analysis apparatuses are provided.
摘要翻译： 核酸分析装置包括多个温度调节装置，旋转机构，输送基座和排出基座，输送驱动机构，排出驱动机构和检测装置。 旋转机构可以包括旋转轴和围绕旋转轴旋转的多个按压部。 反应板组件可以通过按压部分被压在温度调节装置上的状态沿着圆周方向在温度调节装置上移动。 输送驱动机构可以使反应板组件径向向内移动并在按压部分和温度调节装置之间传送。 喷射驱动机构可以使反作用板组件径向向外移动并从压力部分和温度调节装置之间弹出到喷射基座上。 提供反应板，反应板组件和核酸分析装置。
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公开(公告)号：US20090191539A1
公开(公告)日：2009-07-30
申请号：US11976232
申请日：2007-10-23
申请人： Maiko Tanabe , Chihiro Uematsu
发明人： Maiko Tanabe , Chihiro Uematsu
IPC分类号： C12Q1/70
CPC分类号： C12Q1/707
摘要： The present application relates to primers for isothermal amplification of HCV each include at least eighteen consecutive bases corresponding to a 3′ end region of one selected from base sequences of SEQ ID NOs: 1-10, 21 and 22. The primers are specific to HCV subtypes 1a, 1b, 2a, 2b and 3a, respectively and enable genotyping of HCV by isothermal amplification.
摘要翻译： 本申请涉及用于HCV等温扩增的引物各自包括至少十八个连续的碱基，其对应于选自SEQ ID NO：1-10,21和22的碱基序列的3'端区域。引物对HCV 亚型1a，1b，2a，2b和3a分别通过等温扩增进行HCV基因分型。
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公开(公告)号：US20070231808A1
公开(公告)日：2007-10-04
申请号：US11441193
申请日：2006-05-26
IPC分类号： C12Q1/68
CPC分类号： C12Q1/6858 , C12Q1/6818 , C12Q1/6827 , C12Q1/6862 , C12Q2531/125 , C12Q2525/301 , C12Q2521/501 , C12Q2565/107
摘要： This invention provides a method of nucleic acid analysis that enables highly accurate and sensitive quantitation by counting the number of molecules among a plurality of types of genes without amplifying specific genes and that enable reduction of quantitation limits. This method comprises steps of: allowing a polynucleotide comprising a first region having a sequence complementary to the target gene at the 3′ end, a second region having a sequence complementary to the target gene at the 5′ end, and a third region corresponding to a detection probe to hybridize to the target gene; allowing the 3′ end of the first region hybridized to the target gene to ligate to the 5′ end of the second region so as to obtain a circularized polynucleotide; with the use of the circularized polynucleotide as a template, performing a primer extension reaction using a primer having a sequence complementary to part of the circularized polynucleotide and a strand-displacement DNA polymerase; allowing a detection probe containing a sequence identical to the third region to hybridize to a sequence complementary to the third region that iteratively appears in a single-stranded portion of the extension product; and optically detecting the quantity of the detection probe hybridized to the extension product to thereby quantitate the target gene.
摘要翻译： 本发明提供核酸分析方法，通过计数多种类型的基因中的分子数而不扩增特异性基因，并且能够降低定量限度，使得能够进行高度准确和灵敏的定量。 该方法包括以下步骤：允许包含在3'末端具有与靶基因互补的序列的第一区域的多核苷酸，在5'端具有与靶基因互补的序列的第二区域和对应于 与靶基因杂交的检测探针; 允许与靶基因杂交的第一区域的3'末端连接到第二区域的5'端，以获得环化多核苷酸; 使用环化多核苷酸作为模板，使用具有与部分环化多核苷酸互补序列的引物和链置换DNA聚合酶进行引物延伸反应; 允许包含与第三区域相同的序列的检测探针与与重复出现在延伸产物的单链部分中的第三区域互补的序列杂交; 并光学检测与延伸产物杂交的检测探针的量，从而定量靶基因。
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公开(公告)号：US07241597B2
公开(公告)日：2007-07-10
申请号：US10634795
申请日：2003-08-06
申请人： Chihiro Uematsu , Hideki Kambara , Kazunon Okano
发明人： Chihiro Uematsu , Hideki Kambara , Kazunon Okano
CPC分类号： C12Q1/6809 , C12Q1/6855 , C12Q2525/173
摘要： The inventive method for assaying DNA fragments in mixture comprises step 1 of ligating different oligomers hybridizable to primers of the same melting temperature and the same length to individual groups of DNA fragments in a set of DNA fragments; step 2 of mixing together the groups of DNA fragments ligated with the oligomers; step 3 of simultaneous PCR of the groups of DNA fragments ligated with the oligomers in one receptacle by using the primers being complementary to the oligomers and corresponding to the individual groups; and step 4 of detecting PCR amplified DNA fragments; characterized in that the method enables the comparison of plural samples under no influence of PCR reproducibility.
摘要翻译： 用于测定混合物中DNA片段的本发明方法包括步骤1，将不同的低聚物连接到一组DNA片段中与相同融合温度和相同长度的引物相同的DNA片段的单个组; 步骤2，将与低聚物连接的DNA片段组合在一起; 通过使用与寡聚体互补并对应于各个基团的引物，在一个容器中与低聚物连接的DNA片段组同时PCR的步骤3; 和检测PCR扩增的DNA片段的步骤4; 其特征在于，该方法能够在不影响PCR再现性的情况下比较多个样品。
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公开(公告)号：US20060223085A1
公开(公告)日：2006-10-05
申请号：US11332527
申请日：2006-01-17
申请人： Maiko Tanabe , Chihiro Uematsu
发明人： Maiko Tanabe , Chihiro Uematsu
CPC分类号： C12P19/34 , C12Q1/6865 , C12Q2525/179 , C12Q2525/143 , C12Q2521/119
摘要： This invention provides a novel method for amplifying and detecting a target gene rapidly with high sensitivity under isothermal conditions. In such method, a sequence to be amplified can be freely designed regardless of the template sequence, an amplified product can be amplified and detected within a short period of time with high sensitivity, and thus, the gene expression level can be determined more easily than is possible with prior art.
摘要翻译： 本发明提供了一种用于在等温条件下以高灵敏度快速扩增和检测靶基因的新方法。 在这种方法中，无论模板序列如何，都可以自由地设计要扩增的序列，可以在较短的时间内以高灵敏度扩增和检测扩增产物，因此可以更容易地确定基因表达水平 对于现有技术是可能的。
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