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    Isothermal nucleic acid amplification method using surfactant
    1.
    发明授权
    Isothermal nucleic acid amplification method using surfactant 有权
    使用表面活性剂的等温核酸扩增方法

    公开(公告)号：US08435741B2

    公开(公告)日：2013-05-07

    申请号：US12179098

    申请日：2008-07-24

    申请人： Hayato Miyoshi ,  Yoshihide Iwaki ,  Toshihiro Mori

    发明人： Hayato Miyoshi ,  Yoshihide Iwaki ,  Toshihiro Mori

    IPC分类号： C12Q1/68 ,  C12P19/34

    CPC分类号： C12Q1/6853 ,  C12Q2531/119 ,  C12Q2527/125 ,  C12Q2521/101

    摘要： An object to be achieved by the present invention is to provide a nucleic acid amplification method by which a nucleic acid can be amplified substantially isothermally using oligonucleotide primers and DNA polymerase capable of strand displacement. The present invention provides a nucleic acid amplification method which comprises performing substantially isothermal incubation of a reaction solution containing at least one type of deoxynucleotide triphosphate, at least one type of DNA polymerase having strand displacement activity, a divalent cation, at least 0.01% or more surfactant, at least two types of oligonucleotide primer, and the nucleic acid fragment as a template so as to perform a polymerase reaction that initiates from the 3′ end of the primer and thus amplifying the nucleic acid fragment.

    摘要翻译： 本发明要实现的目的是提供一种使用寡核苷酸引物和能够进行链置换的DNA聚合酶基本上等温扩增核酸的核酸扩增方法。 本发明提供了一种核酸扩增方法，其包括对含有至少一种类型的脱氧核苷酸三磷酸的反应溶液进行基本上恒温培养，至少一种具有链置换活性的DNA聚合酶，二价阳离子，至少0​​.01％或更多 表面活性剂，至少两种类型的寡核苷酸引物和核酸片段作为模板，从而进行从引物的3'末端引发并因此扩增核酸片段的聚合酶反应。

    Nucleic acid amplification method
    2.
    发明授权
    Nucleic acid amplification method 有权
    核酸扩增法

    公开(公告)号：US08420323B2

    公开(公告)日：2013-04-16

    申请号：US12270706

    申请日：2008-11-13

    申请人： Hayato Miyoshi ,  Yoshihide Iwaki ,  Toshihiro Mori

    发明人： Hayato Miyoshi ,  Yoshihide Iwaki ,  Toshihiro Mori

    IPC分类号： C12Q1/68 ,  C12P19/34

    CPC分类号： C12Q1/6844 ,  C12Q2531/119 ,  C12Q2525/155 ,  C12Q2525/161

    摘要： An object to be achieved by the present invention is to provide a nucleic acid amplification method by which a nucleic acid can be amplified using oligonucleotide primers and DNA polymerase. The present invention provides a nucleic acid amplification method which comprises performing incubation of a reaction solution containing at least one type of deoxynucleotide triphosphate, at least one type of DNA polymerase, at least two types of oligonucleotide primer, and the nucleic acid fragment as a template so as to perform a polymerase reaction that initiates from the 3′ end of the primer and thus amplifying the nucleic acid fragment, wherein a tag sequence is added at the 5′ end of the first oligonucleotide primer, and the tag sequence is a nucleotide sequence on the template nucleic acid fragment which is present downstream of the sequence which is substantially complementary with the 3′ end region of the first oligonucleotide primer (a region where the first oligonucleotide is annealed to the template nucleic acid).

    摘要翻译： 本发明要实现的目的是提供使用寡核苷酸引物和DNA聚合酶扩增核酸的核酸扩增方法。 本发明提供核酸扩增方法，其包括对含有至少一种脱氧核苷酸三磷酸，至少一种DNA聚合酶，至少两种类型的寡核苷酸引物和核酸片段作为模板的反应溶液进行温育 从而进行从引物的3'端引发的聚合酶反应，从而扩增核酸片段，其中在第一寡核苷酸引物的5'末端添加标签序列，并且标签序列是核苷酸序列 在与第一寡核苷酸引物的3'末端区域（第一寡核苷酸与模板核酸退火的区域）基本上互补的序列下游的模板核酸片段上。

    METHOD FOR DETECTING BACTERIA OF THE GENUS MYCOBACTERIUM (ACID-FAST BACTERIA) AND KIT FOR THE SAME
    3.
    发明申请
    METHOD FOR DETECTING BACTERIA OF THE GENUS MYCOBACTERIUM (ACID-FAST BACTERIA) AND KIT FOR THE SAME 审中-公开
    用于检测一般马铃薯（酸性细菌）细菌的菌株及其相关试剂的方法

    公开(公告)号：US20090023141A1

    公开(公告)日：2009-01-22

    申请号：US11872610

    申请日：2007-10-15

    申请人： Yoshihide Iwaki

    发明人： Yoshihide Iwaki

    IPC分类号： C12Q1/68

    CPC分类号： C12Q1/689

    摘要： An object of the present invention is to provide an oligonucleotide for rapidly and conveniently detecting bacteria of the genus Mycobacterium (acid-fast bacteria) or for identifying the bacterial species thereof, and a method and kit for detecting bacteria of the genus Mycobacterium (acid-fast bacteria) using such oligonucleotid. The present invention provides a method for identifying Mycobacterium tuberculosis, which comprises performing a nucleic acid amplification reaction using a primer for nucleic acid amplification that comprises a nucleotide sequence corresponding to a variable region in a 16S rRNA gene sequence of Mycobacterium tuberculosis and has at least 3 continuous nucleotides contained in the nucleotide sequence represented by SEQ ID NO: 1 at the 3′ end.

    摘要翻译： 本发明的目的是提供一种用于快速方便地检测分枝杆菌属（耐酸菌）的细菌或用于鉴定其细菌种类的寡核苷酸，以及用于检测分枝杆菌属（酸性细菌）的细菌的方法和试剂盒， 快速细菌）使用这种寡核苷酸。 本发明提供了一种用于鉴定结核分枝杆菌的方法，其包括使用核酸扩增引物进行核酸扩增反应，所述引物包含对应于结核分枝杆菌16S rRNA基因序列中的可变区的核苷酸序列，并且具有至少3 包含在3'端由SEQ ID NO：1表示的核苷酸序列中的连续核苷酸。

    Detection method of SNPs
    4.
    发明申请
    Detection method of SNPs 审中-公开
    SNP检测方法

    公开(公告)号：US20080268444A1

    公开(公告)日：2008-10-30

    申请号：US11785203

    申请日：2007-04-16

    申请人： Toshihiro Mori ,  Yoshihide Iwaki

    发明人： Toshihiro Mori ,  Yoshihide Iwaki

    IPC分类号： C12Q1/68

    CPC分类号： C12Q1/6825 ,  C12Q1/6827 ,  C12Q2563/113 ,  C12Q2521/514 ,  C12Q2563/173 ,  C12Q2565/607

    摘要： A method for detecting a mismatch between a target nucleic acid as a measuring object and a control nucleic acid, the method comprising: (a) effecting formation of a double-stranded nucleic acid through hybridization of the control nucleic acid and the target nucleic acid; (b) allowing a mismatch binding protein to contact with the double-stranded nucleic acid and thereby to bind to a mismatched site; (c) allowing an intercalating agent which specifically recognizes the double-stranded nucleic acid and is intercalated therein, to contact with the double-stranded nucleic acid; (d) detecting the intercalating agent intercalated into the double-stranded nucleic acid; and (e) judging the presence or absence of a mismatch between the control nucleic acid and the target nucleic acid, by comparing amounts of the intercalating agent intercalated into the double-stranded nucleic acid in the absence and presence of the mismatch binding protein.

    摘要翻译： 一种用于检测作为测量对象的靶核酸与对照核酸之间的错配的方法，所述方法包括：（a）通过所述对照核酸和所述靶核酸的杂交来实现双链核酸的形成; （b）允许错配结合蛋白与双链核酸接触，从而结合错配位点; （c）允许特异性识别双链核酸并插入其中的插入剂与双链核酸接触; （d）检测嵌入到双链核酸中的插入剂; 和（e）通过在不存在和存在不匹配结合蛋白的情况下比较插入到双链核酸中的插入剂的量来判断对照核酸和靶核酸之间是否存在错配。

    MICROFLUIDIC CHIP
    6.
    发明申请
    MICROFLUIDIC CHIP 审中-公开
    微流感芯片

    公开(公告)号：US20080153152A1

    公开(公告)日：2008-06-26

    申请号：US11943380

    申请日：2007-11-20

    申请人： Akira Wakabayashi ,  Hideyuki Karaki ,  Yoshihiro Sawayashiki ,  Kota Kato ,  Yoshihide Iwaki

    发明人： Akira Wakabayashi ,  Hideyuki Karaki ,  Yoshihiro Sawayashiki ,  Kota Kato ,  Yoshihide Iwaki

    IPC分类号： C12M1/00

    CPC分类号： B01F1/0022 ,  B01F5/0646 ,  B01F5/0655 ,  B01F13/0059 ,  B01F13/1013 ,  B01F13/1027 ,  B01L3/502707 ,  B01L3/502723 ,  B01L7/525 ,  B01L2200/0684 ,  B01L2200/10 ,  B01L2200/16 ,  B01L2300/0654 ,  B01L2300/0816 ,  B01L2300/0864 ,  B01L2300/0867 ,  B01L2400/0445 ,  B01L2400/0487 ,  B01L2400/0688

    摘要： A microfluidic chip, includes: a first port for inputting: a sample liquid; and a first liquid; a second port for inputting a second liquid; a third port for supplying air pressure; a first channel (A) for mixing the sample liquid and the first liquid to generate a first mixed liquid; a second channel (B) for beating the first mixed liquid; a third channel (C) for allowing the second liquid to converge into the first mixed liquid to generate a second mixed liquid; a fourth channel (D) installing a first solid; a fifth channel (E) for promoting mixing of the first solid; a plurality of sixth channels (F) each having a second solid; and a seventh channel (G), which connects the fifth channel (E) and the plurality of sixth channels (F), for dispensing a fixed quantity of the second mixed liquid to each of the plurality of sixth channels (F).

    摘要翻译： 一种微流体芯片，包括：第一端口，用于输入：样品液体; 和第一液体; 用于输入第二液体的第二端口; 用于提供空气压力的第三个端口; 用于混合样品液体和第一液体以产生第一混合液体的第一通道（A）; 用于打浆第一混合液体的第二通道（B）; 第三通道（C），用于允许第二液体会聚到第一混合液体中以产生第二混合液体; 第四通道（D），安装第一固体; 用于促进第一固体的混合的第五通道（E）; 多个第六通道（F），每个具有第二固体; 以及连接第五通道（E）和多个第六通道（F）的第七通道（G），用于将固定量的第二混合液体分配到多个第六通道（F）中的每一个。

    Method for separating and purifying nucleic acid
    9.
    发明申请
    Method for separating and purifying nucleic acid 审中-公开
    核酸分离纯化方法

    公开(公告)号：US20060051799A1

    公开(公告)日：2006-03-09

    申请号：US11217339

    申请日：2005-09-02

    申请人： Yoshihide Iwaki ,  Toshihiro Mori

    发明人： Yoshihide Iwaki ,  Toshihiro Mori

    IPC分类号： C12Q1/68 ,  C12N1/08 ,  C07H21/04

    CPC分类号： C12N15/1006

    摘要： Nucleic acid contained in a sample is highly efficiently recovered at a high recovery ratio by a method for separating and purifying nucleic acid using whole blood as the sample, which is a method for separating and purifying nucleic acid, comprising: preparing a sample solution containing nucleic acid; putting the sample solution containing nucleic acid in contact with a solid phase to allow nucleic acid to be adsorbed to the solid phase; putting a washing solution in contact with the solid phase to wash the solid phase at the state of nucleic acid adsorbed thereon; and putting a elution solution in contact with the solid phase to allow nucleic acid to be desorbed from the solid phase, wherein the step of preparing a sample solution containing nucleic acid comprises at least one selected from the group consisting of vortexing, mixing with inversion, and pipetting.

    摘要翻译： 包含在样品中的核酸通过使用全血作为样品的分离和纯化核酸的方法以高回收率被高效地回收，作为分离和纯化核酸的方法，包括：制备含有核酸的样品溶液 酸; 将含有核酸的样品溶液与固相接触以允许核酸被吸附到固相上; 将洗涤溶液与固相接触以在其上吸附的核酸的状态下洗涤固相; 并将洗脱溶液与固相接触以使核酸从固相中解吸，其中制备含有核酸的样品溶液的步骤包括选自涡旋，与反相混合的至少一种， 和移液。

    Method for analyzing a target nucleic acid fragment and a kit for analyzing a target nucleic acid fragment
    10.
    发明申请
    Method for analyzing a target nucleic acid fragment and a kit for analyzing a target nucleic acid fragment 审中-公开
    用于分析靶核酸片段的方法和用于分析靶核酸片段的试剂盒

    公开(公告)号：US20050266450A1

    公开(公告)日：2005-12-01

    申请号：US11106612

    申请日：2005-04-15

    申请人： Yoshihiko Makino ,  Toshihiro Mori ,  Yoshihide Iwaki

    发明人： Yoshihiko Makino ,  Toshihiro Mori ,  Yoshihide Iwaki

    IPC分类号： C12P19/34 ,  C12Q1/42 ,  C12Q1/68 ,  G01N33/52

    CPC分类号： G01N33/523 ,  C12Q1/42 ,  C12Q1/6816 ,  C12Q1/6869 ,  C12Q2565/625 ,  C12Q2565/301 ,  C12Q2521/543 ,  C12Q2533/101

    摘要： An object of the present invention is to provide a method for analyzing a target nucleic acid fragment which can be simply and swiftly carried out by using a small apparatus, a kit for analyzing a target nucleic acid fragment using the method for analysis, and a dry analytical element for quantifying pyrophosphoric acid. The present invention provides a method for analyzing pyrophosphoric acid generated upon polymerase elongation reaction based on certain nucleotide sequence of a target nucleic acid, a kit for analysis for carrying out the above mentioned method for analysis, and a dry analytical element for quantifying pyrophosphoric acid.

    摘要翻译： 本发明的目的是提供一种用于分析目标核酸片段的方法，其可以通过使用小型装置简单快速地进行，使用该分析方法分析靶核酸片段的试剂盒和干燥的 用于定量焦磷酸的分析元素。 本发明提供一种基于靶核酸的某些核苷酸序列分析聚合酶延伸反应产生的焦磷酸的方法，用于进行上述分析方法的分析用试剂盒和定量焦磷酸的干燥分析元素。