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1.发明授权Pretreatment method of biological sample, detection method of RNA, and pretreatment kit 有权生物样品预处理方法，RNA检测方法及预处理试剂盒公开(公告)号：US09518901B2
公开(公告)日：2016-12-13
申请号：US13807605
申请日：2012-06-28
申请人： Yoshihide Hayashizaki , Kengo Usui , Saori Goda , Kazuhito Nomura , Yuki Kawai
发明人： Yoshihide Hayashizaki , Kengo Usui , Saori Goda , Kazuhito Nomura , Yuki Kawai
CPC分类号： G01N1/34 , C12N15/1003 , C12Q1/6806
摘要： The present invention is to provide a pretreatment method that allows RNA to be detected promptly and simply. RNA degradation activity due to lactoferrin present in the human rhinal mucosa is inhibited, for example, by adding iron ion and carbonate ion to a biological sample that contains the human rhinal mucosa. With the pretreated biological sample, an RNA virus gene can be amplified by a reverse transcriptase. Iron ion and carbonate ion can also inhibit reverse transcriptase inhibition due to lysozyme C contained in the human rhinal mucosa. Further, it is preferable to remove the envelope of the RNA virus by adding SDS to the biological sample that contains the human rhinal mucosa.
摘要翻译： 本发明提供一种能够及时且简单地检测RNA的预处理方法。 例如，通过将铁离子和碳酸根离子加入到含有人类鼻粘膜的生物样品中，可抑制由于存在于人类鼻粘膜中的乳铁蛋白引起的RNA降解活性。 通过预处理的生物样品，RNA病毒基因可以通过逆转录酶扩增。 铁离子和碳酸根离子也可以抑制人类鼻粘膜中包含的溶菌酶C的逆转录酶抑制。 此外，优选通过向包含人类鼻粘膜的生物样品中加入SDS来除去RNA病毒的包膜。
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2.发明申请PRETREATMENT METHOD OF BIOLOGICAL SAMPLE, DETECTION METHOD OF RNA, AND PRETREATMENT KIT 有权生物样品的预处理方法，RNA的检测方法和预处理试剂盒公开(公告)号：US20130302783A1
公开(公告)日：2013-11-14
申请号：US13807605
申请日：2012-06-28
申请人： Yoshihide Hayashizaki , Kengo Usui , Saori Goda , Kazuhito Nomura , Yuki Kawai
发明人： Yoshihide Hayashizaki , Kengo Usui , Saori Goda , Kazuhito Nomura , Yuki Kawai
IPC分类号： G01N1/34
CPC分类号： G01N1/34 , C12N15/1003 , C12Q1/6806
摘要： The present invention is to provide a pretreatment method that allows RNA to be detected promptly and simply. RNA degradation activity due to lactoferrin present in the human rhinal mucosa is inhibited, for example, by adding iron ion and carbonate ion to a biological sample that contains the human rhinal mucosa. With the pretreated biological sample, an RNA virus gene can be amplified by a reverse transcriptase. Iron ion and carbonate ion can also inhibit reverse transcriptase inhibition due to lysozyme C contained in the human rhinal mucosa. Further, it is preferable to remove the envelope of the RNA virus by adding SDS to the biological sample that contains the human rhinal mucosa.
摘要翻译： 本发明提供一种能够及时且简单地检测RNA的预处理方法。 例如，通过将铁离子和碳酸根离子加入到含有人类鼻粘膜的生物样品中，可抑制由于存在于人类鼻粘膜中的乳铁蛋白引起的RNA降解活性。 通过预处理的生物样品，RNA病毒基因可以通过逆转录酶扩增。 铁离子和碳酸根离子也可以抑制人类鼻粘膜中包含的溶菌酶C的逆转录酶抑制。 此外，优选通过向包含人类鼻粘膜的生物样品中加入SDS来除去RNA病毒的包膜。
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公开(公告)号：US20110236900A1
公开(公告)日：2011-09-29
申请号：US13130993
申请日：2009-11-27
CPC分类号： C07K14/195 , G01N33/5308 , G01N2333/32
摘要： A method for determining the presence or absence of a mutation on the basis of the presence or absence of amplification with high reliability is provided. A target sequence including a target site contained in a sample nucleic acid is amplified using a primer that can hybridize to a region including the target site contained in the sample nucleic acid in the presence of a novel MutS having an amino acid sequence of SEQ ID NO: 2, and then the presence or absence of a mutation at the target site is determined on the basis of the presence or absence of amplification. The novel MutS binds more specifically to a mismatched base pair than to a fully-matched base pair, whereby an extension reaction caused by a mismatch-binding primer is suppressed. Thus, according to the present invention, the presence or absence of a mutation can be determined with high reliability.
摘要翻译： 提供了一种基于高可靠性的扩增的存在或不存在来确定突变的存在或不存在的方法。 包含样品核酸中包含的靶位点的靶序列使用可以在含有氨基酸序列SEQ ID NO的新型MutS的存在下与包含在样品核酸中的靶位点的区域杂交的引物进行扩增 ：2，然后基于扩增的存在或不存在来确定靶位点处的突变的存在或不存在。 新的MutS更具体地结合失配的碱基对而不是完全匹配的碱基对，由此抑制由错配结合引物引起的延伸反应。 因此，根据本发明，可以高可靠性确定突变的存在或不存在。
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公开(公告)号：US09586987B2
公开(公告)日：2017-03-07
申请号：US14343511
申请日：2012-09-07
申请人： Yoshihide Hayashizaki , Yasumasa Kimura , Kengo Usui , Yuki Tanaka , Yuki Kawai
发明人： Yoshihide Hayashizaki , Yasumasa Kimura , Kengo Usui , Yuki Tanaka , Yuki Kawai
CPC分类号： C07H21/04 , C12Q1/6844 , C12Q1/6853 , C12Q2525/155 , C12Q2525/301 , C12Q2531/119 , C12Q2531/125 , C12Q2525/161 , C12Q2527/101
摘要： The present invention provides a primer set including primers that can be designed easily, with which an amplification distance can be shortened. Provided is a primer set for use in a method for isothermally amplifying a target nucleic acid sequence 4. The primer set includes a first primer 1F and a second primer 1R. The first primer 1F includes, on the 3′ side thereof, a sequence (A′) that can hybridize to a sequence (A) on the 3′ side of the target nucleic acid sequence. The second primer 1R includes, on the 3′ side thereof, a sequence (B′) that can hybridize to a sequence (B) on the 3′ side of either a strand extended from the first primer or a complementary strand of the target nucleic acid sequence 4. The first primer 1F and the second primer 1R include, on the 5′ sides thereof, sequences (C) that are substantially identical to each other.
摘要翻译： 本发明提供了可以容易地设计的引物，其可以缩短放大距离。 提供了用于等温扩增靶核酸序列4的方法中的引物组。引物组包括第一引物1F和第二引物1R。 第一引物1F在其3'侧包含可以与靶核酸序列的3'侧的序列（A）杂交的序列（A'）。 第二引物1R在其3'侧具有可以与从第一引物延伸的链的3'侧的序列（B）或靶核酸的互补链上的序列（B'）杂交的序列 酸序列4.第一引物1F和第二引物1R在其5'侧包括彼此基本相同的序列（C）。
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5.发明申请PRIMER SET, METHOD FOR AMPLIFYING TARGET NUCLEIC ACID SEQUENCE USING SAME, AND METHOD FOR DETECTING MUTATED NUCLEIC ACID USING SAME 有权引物组，使用其放大靶核酸序列的方法和使用其检测突变的核酸的方法公开(公告)号：US20140295447A1
公开(公告)日：2014-10-02
申请号：US14343511
申请日：2012-09-07
申请人： Yoshihide Hayashizaki , Yasumasa Kimura , Kengo Usui , Yuki Tanaka , Yuki Kawai
发明人： Yoshihide Hayashizaki , Yasumasa Kimura , Kengo Usui , Yuki Tanaka , Yuki Kawai
CPC分类号： C07H21/04 , C12Q1/6844 , C12Q1/6853 , C12Q2525/155 , C12Q2525/301 , C12Q2531/119 , C12Q2531/125 , C12Q2525/161 , C12Q2527/101
摘要： The present invention provides a primer set including primers that can be designed easily, with which an amplification distance can be shortened. Provided is a primer set for use in a method for isothermally amplifying a target nucleic acid sequence 4. The primer set includes a first primer 1F and a second primer 1R. The first primer 1F includes, on the 3′ side thereof, a sequence (A′) that can hybridize to a sequence (A) on the 3′ side of the target nucleic acid sequence. The second primer 1R includes, on the 3′ side thereof, a sequence (B′) that can hybridize to a sequence (B) on the 3′ side of either a strand extended from the first primer or a complementary strand of the target nucleic acid sequence 4. The first primer 1F and the second primer 1R include, on the 5′ sides thereof, sequences (C) that are substantially identical to each other.
摘要翻译： 本发明提供了可以容易地设计的引物，其可以缩短放大距离。 提供了用于等温扩增靶核酸序列4的方法中的引物组。引物组包括第一引物1F和第二引物1R。 第一引物1F在其3'侧包含可以与靶核酸序列的3'侧的序列（A）杂交的序列（A'）。 第二引物1R在其3'侧具有可以与从第一引物延伸的链的3'侧的序列（B）或靶核酸的互补链上的序列（B'）杂交的序列 酸序列4.第一引物1F和第二引物1R在其5'侧包括彼此基本相同的序列（C）。
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公开(公告)号：US08097414B2
公开(公告)日：2012-01-17
申请号：US12094896
申请日：2006-11-21
IPC分类号： C12P19/34
CPC分类号： C12Q1/6844 , C12Q1/6865 , C12Q2527/125 , C12Q2525/301
摘要： Problem to be solved There is provided a method for detecting and/or amplifying a nucleic acid contained in a biological sample such as blood or cells conveniently, rapidly, and effectively.Solution There is provided a method for detecting a nucleic acid contained in a sample, comprising the step of adding at least one substance selected from the group consisting of polyphenols, polyhydric alcohols, sugar acids, sugar alcohols, and hydrophilic biodegradable polymers to a sample, the step of complementarily binding an oligonucleotide complementary to a part of the nucleic acid sequence of a nucleic acid to be detected to a part of the nucleic acid sequence, and the step of detecting the nucleic acid to be detected.
摘要翻译： 待解决的问题提供一种方便，快速，有效地检测和/或扩增生物样品如血液或细胞中的核酸的方法。 溶液提供了一种检测样品中所含的核酸的方法，其包括将至少一种选自多酚，多元醇，糖酸，糖醇和亲水性可生物降解聚合物的物质加入到样品中的步骤， 将与要检测的核酸的一部分核酸序列互补的寡核苷酸互补结合到核酸序列的一部分的步骤和检测待检测的核酸的步骤。
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公开(公告)号：US20090042197A1
公开(公告)日：2009-02-12
申请号：US12094896
申请日：2006-11-21
CPC分类号： C12Q1/6844 , C12Q1/6865 , C12Q2527/125 , C12Q2525/301
摘要： Problem to be Solved There is provided a method for detecting and/or amplifying a nucleic acid contained in a biological sample such as blood or cells conveniently, rapidly, and effectively.Solution There is provided a method for detecting a nucleic acid contained in a sample, comprising the step of adding at least one substance selected from the group consisting of polyphenols, polyhydric alcohols, sugar acids, sugar alcohols, and hydrophilic biodegradable polymers to a sample, the step of complementarily binding an oligonucleotide complementary to a part of the nucleic acid sequence of a nucleic acid to be detected to a part of the nucleic acid sequence, and the step of detecting the nucleic acid to be detected.
摘要翻译： 待解决的问题提供了一种方便，快速，有效地检测和/或扩增生物样品如血液或细胞中的核酸的方法。 溶液提供了一种检测样品中所含的核酸的方法，其包括将至少一种选自多酚，多元醇，糖酸，糖醇和亲水性可生物降解聚合物的物质加入到样品中的步骤， 将与要检测的核酸的一部分核酸序列互补的寡核苷酸互补结合到核酸序列的一部分的步骤和检测待检测的核酸的步骤。
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8.发明授权公开(公告)号：US08017320B2
公开(公告)日：2011-09-13
申请号：US10554678
申请日：2008-05-14
IPC分类号： C12Q1/68
CPC分类号： G01N33/57446 , C12Q1/6886 , C12Q2600/112 , G01N2333/90203 , G01N2333/988 , G01N2800/56
摘要： Metastatic cancer cells originating from gastric cancer are detected by a method comprising the step of collecting a biological sample from a subject, the step of detecting the presence of at least either aldehyde dehydrogenase or dopa decarboxylase in the biological sample of the subject, and the step of determining that the possibility of containing metastatic cancer cells originating from the gastric cancer in the sample is high when at least either aldehyde dehydrogenase or dopa decarboxylase is present. By the use of these as markers for metastatic cancer cells originating from gastric cancer, the presence or absence of peritoneal metastasis in a gastric cancer patient can be detected rapidly and reliably, and data important for deciding whether intraperitoneal cancer chemotherapy should be applied is provided.
摘要翻译： 通过包括从受试者收集生物样品的步骤的方法检测源自胃癌的转移性癌细胞，检测受试者的生物样品中至少一种醛脱氢酶或多巴脱羧酶的存在的步骤，以及步骤 确定当存在至少一种醛脱氢酶或多巴脱羧酶时，含有样品中源自胃癌的转移性癌细胞的可能性高。 通过使用这些作为来自胃癌的转移性癌细胞的标记物，可以快速可靠地检测胃癌患者腹膜转移的存在或不存在，并且提供了决定是否应用腹膜内癌化疗的重要数据。
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公开(公告)号：US20100035249A1
公开(公告)日：2010-02-11
申请号：US12186009
申请日：2008-08-05
申请人： Yoshihide Hayashizaki , Piero Carninci , Mutsumi Katayama , Shintaro Katayama , Masayoshi Itoh , Matthias Harbers
发明人： Yoshihide Hayashizaki , Piero Carninci , Mutsumi Katayama , Shintaro Katayama , Masayoshi Itoh , Matthias Harbers
IPC分类号： C12Q1/68
CPC分类号： C12Q1/6809 , C12Q1/6834 , C12Q1/6874 , C12Q2535/101 , C12Q2565/537 , C12Q2525/155
摘要： The present invention provides methods for the sequencing of all RNA species within an RNA sample, such as the RNA content obtained from a cell, a tissue, a living organism, or from an artificial source. RNA molecules within the samples are labeled in a RNA-specific manner prior to immobilization on a solid support. One label is used to mark the location of the RNA molecule on the solid support, whereas the second label is used to mark selectively the S′ end of full-length mRNA molecules. RNA molecules are sequenced while being bound to the solid support in one or more sequencing reactions, and sequences of individual RNA molecules can be forwarded to computational analysis for assembling sequence information from individual sequencing reads obtained from the same location on the solid support. Not only unsupervised expression profiling on a genome-wide scale, but also the direct analysis of RNA-RNA interactions become possible as revealed by the analysis of the sequencing information obtained along with genomic information.
摘要翻译： 本发明提供了用于对RNA样品中所有RNA物种进行测序的方法，例如从细胞，组织，活生物体或人造来源获得的RNA含量。 在固定在固体支持物上之前，将样品中的RNA分子以RNA特异性方式标记。 一个标签用于标记RNA分子在固体支持物上的位置，而第二个标记用于选择性标记全长mRNA分子的S'末端。 RNA分子在一个或多个测序反应中与固体支持物结合时进行测序，并且可将单个RNA分子的序列转发到计算分析，以从固体支持物上的相同位置获得的单独测序读数组装序列信息。 不仅在基因组范围内无监督的表达谱，而且通过分析与基因组信息一起获得的测序信息显示，RNA-RNA相互作用的直接分析也是可能的。
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公开(公告)号：US20060160084A1
公开(公告)日：2006-07-20
申请号：US10532975
申请日：2003-10-29
CPC分类号： C12Q1/6844 , C12Q2525/301
摘要： The present invention relates to a process for synthesizing or amplifying efficiently a nucleic acid comprising a target nucleic acid sequence. In the process according to the present invention, a primer comprising in its 3′-end portion a sequence (Ac′) which hybridizes a sequence (A) in the 3′-end portion of the target nucleic acid sequence, and in the 5′-side of said sequence (Ac′) a sequence (B′) which hybridizes the complementary sequence (Bc) of a sequence (B) positioned in the 5′-side of said sequence (A) on the target nucleic acid sequence, wherein {X−(Y−Y′)}/X is in the range of −1.00 to 1.00, in which X denotes the number of bases in said sequence (Ac′), Y denotes the number of bases in the region flanked by said sequences (A) and (B) in the target nucleic acid sequence, and Y′ denotes the number of bases in an intervening sequence between said sequences (Ac′) and (B′) (Y′ may be zero).
摘要翻译： 本发明涉及有效地合成或扩增包含靶核酸序列的核酸的方法。 在本发明的方法中，引物在其3'末端部分包含与目标核酸序列的3'末端部分的序列（A）杂交的序列（Ac'）和5 所述序列（Ac'）的序列（B'）与位于靶核酸序列上的所述序列（A）的5'侧的序列（B）的互补序列（Bc）杂交， 其中{X-（Y-Y'）} / X在-1.00至1.00的范围内，其中X表示所述序列中的碱基数（Ac'），Y表示侧翼的区域中的碱基数 所述序列（A）和（B）在靶核酸序列中，Y'表示所述序列（Ac'）和（B'）之间的间插序列中的碱基数（Y'可以为零）。
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