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    METHOD FOR DISTINGUISHING TARGET BASE SEQUENCE
    1.
    发明公开
    METHOD FOR DISTINGUISHING TARGET BASE SEQUENCE 有权
    VERFAHREN ZUR UNTERSCHEIDUNG VON ZIELBASISSEQUENZEN

    公开(公告)号：EP2554680A1

    公开(公告)日：2013-02-06

    申请号：EP11762732.3

    申请日：2011-03-25

    申请人： Toppan Printing Co., Ltd. ,  Kyushu University, National University Corporation

    发明人： KITANO, Shiro ,  YAMANE, Akio ,  MARUYAMA, Atsushi

    IPC分类号： C12Q1/68 ,  C12N15/09 ,  G01N21/78 ,  G01N33/53 ,  G01N33/533

    CPC分类号： C12Q1/6832 ,  C12Q1/6827 ,  C12Q2527/125 ,  C12Q2537/1373 ,  C12Q2545/107

    摘要： According to the present invention, a method for identifying a base sequence accompanying competitive hybridization is provided, wherein a nucleic acid having a target base sequence can be identified in an extremely short period of time as compared with conventional methods while maintaining the accuracy of identifying differences in base sequences. The method of the present invention comprises a thermal denaturation step for subjecting a sample double-stranded nucleic acid and a reference double-stranded nucleic acid containing the same base sequence as a target base sequence to thermal denaturation treatment in a single reaction solution, a temperature lowering step for carrying out competitive hybridization between the sample double-stranded nucleic acid and the reference double-stranded nucleic acid by lowering the temperature of the reaction solution after the thermal denaturation step, a measurement step for measuring a double-stranded nucleic acid formed by a nucleic acid strand that composed the reference double-stranded nucleic acid and a nucleic acid strand that composed the sample double-stranded nucleic acid, and an identification step for identifying identity between the reference double-stranded nucleic acid and the sample double-stranded nucleic acid based on measurement results obtained from the measurement step; wherein, the temperature lowering step is carried out in the presence of a cationic comb-type polymer.

    摘要翻译： 根据本发明，提供了用于鉴定伴随竞争性杂交的碱基序列的方法，其中与常规方法相比，可以在极短的时间内鉴定具有目标碱基序列的核酸，同时保持识别差异的准确性 在碱基序列中。 本发明的方法包括热变性步骤，用于将单个反应溶液中的样品双链核酸和含有相同碱基序列的参考双链核酸作为目标碱基序列进行热变性处理，温度 通过降低热变性步骤后的反应溶液的温度来进行样品双链核酸与参考双链核酸之间的竞争性杂交的降低步骤，测定由双链核酸形成的双链核酸的测定步骤 构成参考双链核酸的核酸链和构成样品双链核酸的核酸链，以及用于鉴定参考双链核酸和样品双链核酸之间的同一性的鉴定步骤 基于从测量步骤获得的测量结果; 其中，降温步骤在阳离子梳型聚合物的存在下进行。

    METHOD FOR DETECTING TARGET BASE SEQUENCE USING COMPETITIVE PRIMER
    5.
    发明公开
    METHOD FOR DETECTING TARGET BASE SEQUENCE USING COMPETITIVE PRIMER 有权
    VERFAHREN ZUR ERKENNUNG VON ZIELBASISSEQUENZEN MITHILFE KONKURRIERENDER PRIMER

    公开(公告)号：EP2551356A1

    公开(公告)日：2013-01-30

    申请号：EP11759408.5

    申请日：2011-03-23

    申请人： Toppan Printing Co., Ltd.

    发明人： MAKINO, Yoichi ,  YAMANE, Akio ,  NAKAYAMA, Masato

    IPC分类号： C12Q1/68 ,  C12N15/09

    CPC分类号： C12Q1/6869 ,  C12Q1/6827 ,  C12Q1/6858 ,  C12Q2531/119 ,  C12Q2537/161 ,  C12Q2531/143 ,  C12Q2531/113 ,  C12Q2525/161 ,  C12Q2525/185 ,  C12Q2525/204 ,  C12Q2535/125 ,  C12Q2549/101 ,  C12Q2563/107 ,  C12Q2565/101

    摘要： Disclosed is a method of detecting a target base sequence having a polymorphic base, the method including: (a) a step of adding to a nucleic acid sample having a target nucleic acid that includes a base sequence including the target base sequence: at least one type of detection primer, at least one type of competitive primer, and at least one type of common primer; (b) a step of annealing the detection primer and the competitive primer to the target nucleic acid in a competitive manner, thereby synthesizing an extension product A; (c) a step of annealing the common primer to the extension product A obtained in the step (b) or in the following step (d), thereby synthesizing an extension product B; (d) a step of annealing the detection primer or the competitive primer to the extension product B obtained in the previous step (c), thereby synthesizing the extension product A; and (e) a step of detecting the extension product A or the extension product B.

    摘要翻译： 本发明公开了一种检测具有多态碱基的目标碱基序列的方法，该方法包括：（a）向具有包含目标碱基序列的碱基序列的靶核酸的核酸样品中添加至少一种 检测引物的类型，至少一种类型的竞争性引物和至少一种类型的共同引物; （b）以竞争的方式将检测引物和竞争性引物退火至靶核酸，由此合成延伸产物A; （c）将通用引物退火到步骤（b）或后续步骤（d）中获得的延伸产物A的步骤，从而合成延伸产物B; （d）将检测引物或竞争性引物退火到在前面步骤（c）中获得的延伸产物B的步骤，从而合成延伸产物A; 和（e）检测扩展产品A或扩展产品B的步骤。

    METHOD OF DISTINGUISHING GENOTYPES
    6.
    发明公开
    METHOD OF DISTINGUISHING GENOTYPES 有权
    鉴别基因型的方法

    公开(公告)号：EP2418289A1

    公开(公告)日：2012-02-15

    申请号：EP10758232.2

    申请日：2010-03-26

    申请人： Toppan Printing Co., Ltd.

    发明人： YAMANE, Akio ,  NAKAYAMA, Mashimo ,  KITANO, Shiro

    IPC分类号： C12Q1/68 ,  C12N15/09 ,  G01N21/78 ,  G01N33/50 ,  G01N33/566

    CPC分类号： C12Q1/686 ,  C12Q1/6858 ,  G01N2021/6441 ,  C12Q2537/161 ,  C12Q2535/131 ,  C12Q2531/119 ,  C12Q2525/186 ,  C12Q2527/125

    摘要： The present invention relates to a method of distinguishing genotypes using PCR-PHFA including: a nucleic acid amplification step in which a mutation site-including region of a gene contained in a specimen is amplified by a nucleic acid amplification reaction, thereby obtaining an amplification reaction solution which contains a specimen double-stranded nucleic acid; and a distinction step in which the amplification reaction solution obtained from the nucleic acid amplification step is mixed with a reference double-stranded nucleic acid having a specific genotype on the mutation site as well as being labeled with a labeling substance, and the mixture is subjected to a competitive strand displacement reaction, and the level of the occurrence of strand displacement between the reference double-stranded nucleic acid and the specimen double-stranded nucleic acid is assessed so as to distinguish the identity between the reference double-stranded nucleic acid and the specimen double-stranded nucleic acid; and the competitive strand displacement reaction is performed under a condition to suppress a polymerase extension reaction, and a genotype distinguishing kit for use in the distinct of genotypes by this method.

    摘要翻译： 本发明涉及使用PCR-PHFA区分基因型的方法，其包括：核酸扩增步骤，其中通过核酸扩增反应扩增标本中所含基因的突变位点包含区域，从而获得扩增反应 含有标本双链核酸的溶液; 以及将由核酸扩增步骤得到的扩增反应液与在突变位点上具有特定基因型的标准双链核酸混合并用标记物质标记的区分步骤， 进行竞争性链置换反应，并且评估参考双链核酸和样本双链核酸之间链置换发生的水平，以区分参考双链核酸和参考双链核酸之间的同一性 标本双链核酸; 竞争链置换反应在抑制聚合酶延伸反应的条件下进行，并且通过该方法使用基因型不同的基因型判别试剂盒。