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    Method of analyzing methylated DNA
    2.
    发明授权
    Method of analyzing methylated DNA 有权
    分析甲基化DNA的方法

    公开(公告)号：US08163490B2

    公开(公告)日：2012-04-24

    申请号：US12403167

    申请日：2009-03-12

    申请人： Ayako Sakai ,  Masahiro Kajita ,  Hideki Ishihara

    发明人： Ayako Sakai ,  Masahiro Kajita ,  Hideki Ishihara

    IPC分类号： C12Q1/68 ,  C12P19/34

    CPC分类号： C12Q1/6827 ,  C12Q2521/331 ,  C12Q2531/113

    摘要： The present invention provides a method of analyzing methylated DNA, comprising steps of: (A) treating a DNA-containing sample with a restriction enzyme to obtain a sample containing a DNA fragment; (B) concentrating methylated DNA contained in the sample obtained in step (A) to obtain a methylated DNA concentrate; (C) subjecting the methylated DNA concentrate obtained in step (B) and a primer set to nucleic acid amplification reaction, wherein the primer set performs the nucleic acid amplification reaction in step (C) by using a template DNA which does not have a CpG site; (D) detecting an amplification product obtained in step (C); (E) judging whether the methylated DNA concentrate obtained in step (B) is appropriate as a sample for detection of methylated DNA, on the basis of the detection result of the amplification product in step (D); and (F) analyzing the methylated DNA contained in the methylated DNA concentrate.

    摘要翻译： 本发明提供一种分析甲基化DNA的方法，包括以下步骤：（A）用限制酶处理含有DNA的样品，得到含有DNA片段的样品; （B）浓缩步骤（A）中获得的样品中所含的甲基化DNA以获得甲基化DNA浓缩物; （C）使步骤（B）中获得的甲基化DNA浓缩物和引物组进行核酸扩增反应，其中引物组通过使用不具有CpG的模板DNA进行步骤（C）中的核酸扩增反应 现场; （D）检测步骤（C）中获得的扩增产物; （E）基于步骤（D）中的扩增产物的检测结果判断步骤（B）中获得的甲基化DNA浓缩物是否适合作为检测甲基化DNA的样品; 和（F）分析甲基化DNA浓缩物中所含的甲基化DNA。

    Method for confirming conversion treatment and nucleic acid molecule used therefor
    4.
    发明申请
    Method for confirming conversion treatment and nucleic acid molecule used therefor 审中-公开
    用于确认转化处理的方法和用于其的核酸分子

    公开(公告)号：US20080096213A1

    公开(公告)日：2008-04-24

    申请号：US11902254

    申请日：2007-09-20

    申请人： Masahiro Kajita ,  Noriaki Yamamoto ,  Ayako Sakai ,  Hideki Ishihara

    发明人： Masahiro Kajita ,  Noriaki Yamamoto ,  Ayako Sakai ,  Hideki Ishihara

    IPC分类号： C12Q1/68 ,  C07H21/00

    CPC分类号： C12Q1/6827 ,  C12Q2565/1015 ,  C12Q2545/114 ,  C12Q2545/101

    摘要： There is provided a method for confirming conversion treatment of non-methylated cytosine, including a step of mixing a DNA taken from a living body, with a nucleic acid for accuracy control having a sequence A which is not contained in a genome of the living body and contains cytosine, a step of converting non-methylated cytosine in the mixture into a base other than cytosine, a step of detecting the sequence A and/or a sequence A′ which has a sequence obtained by converting cytosine in the sequence A into the base other than cytosine, and a step of determining whether the conversion step has been properly performed or not based on the result of the detection step.

    摘要翻译： 提供了一种确认非甲基化胞嘧啶的转化处理的方法，包括将从活体取得的DNA混合的步骤与用于精确控制的核酸的步骤，所述序列A不包含在生物体的基因组中 并含有胞嘧啶，将混合物中的非甲基化胞嘧啶转化为除胞嘧啶以外的碱基的步骤，检测序列A和/或A序列的步骤，其具有通过将序列A中的胞嘧啶转化为 碱基以外的碱基，以及基于检测步骤的结果确定转化步骤是否已经被适当地进行的步骤。

    METHOD OF DETECTING METHYLATED CYTOSINE
    5.
    发明申请
    METHOD OF DETECTING METHYLATED CYTOSINE 有权
    检测甲基化胞嘧啶的方法

    公开(公告)号：US20100240042A1

    公开(公告)日：2010-09-23

    申请号：US12674648

    申请日：2008-08-20

    申请人： Masahiro Kajita ,  Noriko Oka ,  Noriaki Yamamoto ,  Ayako Sakai ,  Hideki Ishihara

    发明人： Masahiro Kajita ,  Noriko Oka ,  Noriaki Yamamoto ,  Ayako Sakai ,  Hideki Ishihara

    IPC分类号： C12Q1/68 ,  C07H21/04 ,  C12M1/34

    CPC分类号： C12Q1/6827 ,  C12Q2537/164 ,  C12Q2525/119 ,  C12Q2523/115

    摘要： The method for detecting a methylated cytosine of the present invention comprises the steps of:hybridizing a sample DNA with an oligonucleotide which can hybridize with a region of the sample DNA containing a cytosine suspected of being methylated and has an a basic site at the position complementary to the cytosine;reacting the hybridized sample DNA obtained in the previous step with an oxidizing agent to oxidize the cytosine when it is methylated; anddetecting the oxidized methylated cytosine.

    摘要翻译： 检测本发明的甲基化胞嘧啶的方法包括以下步骤：将样品DNA与能够与疑似被甲基化的胞嘧啶的样品DNA的区域杂交的寡核苷酸杂交，并且在互补位置具有基本位点 到胞嘧啶; 使上一步获得的杂交样品DNA与氧化剂反应，以在甲基化时氧化胞嘧啶; 并检测氧化的甲基化胞嘧啶。

    METHOD FOR DETERMINING PRESENCE OR ABSENCE OF EPITHELIAL CANCER-ORIGIN CELL IN BIOLOGICAL SAMPLE, AND MOLECULAR MARKER AND KIT THEREFOR
    7.
    发明申请
    METHOD FOR DETERMINING PRESENCE OR ABSENCE OF EPITHELIAL CANCER-ORIGIN CELL IN BIOLOGICAL SAMPLE, AND MOLECULAR MARKER AND KIT THEREFOR 审中-公开
    用于确定生物样品中上位癌细胞存在或不存在的方法及其分子标记和试剂盒

    公开(公告)号：US20120190024A1

    公开(公告)日：2012-07-26

    申请号：US13381620

    申请日：2010-06-30

    申请人： Kaya Tai ,  Masahiro Kajita ,  Yoshihiko Tashima ,  Ayako Sakai

    发明人： Kaya Tai ,  Masahiro Kajita ,  Yoshihiko Tashima ,  Ayako Sakai

    IPC分类号： C12Q1/68 ,  C07H21/04

    CPC分类号： C12Q1/6886 ,  C12Q2600/154

    摘要： The present invention provides a method for determination of presence or absence of an epithelial cancer-derived cell in a biological sample obtained from a subject comprising the steps of: extracting DNA from the biological sample, analyzing methylation status of a CpG site located in at least one region represented by base sequences SEQ ID NOs: 1, 2, 3 and 4 in the DNA obtained from the step of extracting, and determining presence or absence of the epithelial cancer-derived cell in the biological sample based on an analysis result obtained from the step of analyzing.

    摘要翻译： 本发明提供一种用于确定从受试者获得的生物样品中存在或不存在上皮癌衍生细胞的方法，包括以下步骤：从生物样品中提取DNA，分析至少位于至少一个CpG位点的甲基化状态 由从提取步骤获得的DNA中的碱基序列SEQ ID NO：1,2,3和4表示的一个区域，并且基于从生物样品中获得的分析结果确定生物样品中上皮癌来源的细胞的存在或不存在 分析的步骤。

    Hybridoma producing anti-methylated DNA antibody and utilization of same
    8.
    发明授权
    Hybridoma producing anti-methylated DNA antibody and utilization of same 有权
    产生抗甲基化DNA抗体的杂交瘤及其利用

    公开(公告)号：US09169330B2

    公开(公告)日：2015-10-27

    申请号：US13430295

    申请日：2012-03-26

    申请人： Ayako Sakai ,  Masahiro Kajita

    发明人： Ayako Sakai ,  Masahiro Kajita

    IPC分类号： C07K16/18 ,  C07H1/06 ,  C12N5/16 ,  C07K16/44 ,  G01N33/53

    CPC分类号： C07K16/44 ,  C12N5/163 ,  G01N33/5308

    摘要： The present invention relates to a hybridoma producing an anti-methylated DNA antibody, obtained by cell fusion of an antibody-producing cell obtained from an animal immunized with an antigen containing 5′-(5-methyl-2′-deoxycytidine-3′-phospho)-2′-deoxyguanosine 3′-phosphate with a myeloma cell. The present invention also relates to a monoclonal antibody produced by the hybridoma and a method for immunoprecipitation of a methylated DNA using the antibody.

    摘要翻译： 本发明涉及产生抗甲基化DNA抗体的杂交瘤，该抗体通过细胞融合获得的抗体产生细胞获得，所述抗体产生细胞由含有5' - （5-甲基-2'-脱氧胞苷-3'- 磷酸）-2'-脱氧鸟苷3'-磷酸与骨髓瘤细胞。 本发明还涉及由该杂交瘤产生的单克隆抗体以及使用该抗体对甲基化DNA进行免疫沉淀的方法。

    Kit and method for determining whether or not unmethylated cytosine conversion treatment is properly carried out and method for analyzing methylated DNA using the same
    10.
    发明授权
    Kit and method for determining whether or not unmethylated cytosine conversion treatment is properly carried out and method for analyzing methylated DNA using the same 有权
    用于确定是否正确进行非甲基化胞嘧啶转化处理的试剂盒和方法以及使用其分析甲基化DNA的方法

    公开(公告)号：US08906623B2

    公开(公告)日：2014-12-09

    申请号：US12935846

    申请日：2009-03-16

    申请人： Kaya Tai ,  Ayako Sakai ,  Noriaki Yamamoto ,  Noriko Oka ,  Masahiro Kajita

    发明人： Kaya Tai ,  Ayako Sakai ,  Noriaki Yamamoto ,  Noriko Oka ,  Masahiro Kajita

    IPC分类号： C12Q1/68 ,  C12P19/34

    CPC分类号： C12Q1/6827 ,  C12Q2523/125

    摘要： Success or failure of unmethylated cytosine conversion treatment is determined by using a first primer set comprising plural primers that hybridize with a nucleic acid comprising a nucleotide sequence not containing cytosine in the nucleotide sequence of biological DNA that is subject to an unmethylated cytosine conversion treatment and a second primer set comprising plural primers that hybridize with a nucleic acid comprising a nucleotide sequence in which cytosine in a nucleotide sequence containing cytosine and not containing a CpG site is converted into a base other than cytosine, in the nucleotide sequence of biological DNA.

    摘要翻译： 通过使用包含多个引物的第一引物组来确定未甲基化胞嘧啶转化处理的成功或失败，所述多个引物与包含未经甲基化胞嘧啶转化处理的生物DNA的核苷酸序列中不含胞嘧啶的核苷酸序列的核酸杂交， 第二引物组，其包含与包含核苷酸序列的核酸杂交的多个引物，所述核酸序列在生物DNA的核苷酸序列中将含有胞嘧啶并且不含有CpG位点的核苷酸序列中的胞嘧啶转化为除胞嘧啶以外的碱基。