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1. 发明公开

EP3339447A1 METHOD FOR ANALYZING TEMPLATE NUCLEIC ACID, METHOD FOR ANALYZING TARGET MATERIAL, KIT FOR ANALYZING TEMPLATE NUCLEIC ACID OR TARGET MATERIAL, AND DEVICE FOR ANALYZING TEMPLATE NUCLEIC ACID OR TARGET MATERIAL 有权
公开(公告)号：EP3339447A1
公开(公告)日：2018-06-27
申请号：EP16841714.5
申请日：2016-08-26
申请人： Kabushiki Kaisha DNAFORM
发明人： TANAKA Yuji , HAYASHIZAKI Yoshihide , TSUJIMARU Koichiro
IPC分类号： C12Q1/68 , C12M1/00 , G01N21/64 , C12N15/09
摘要： The present invention provides a method for analyzing a template nucleic acid, a method for analyzing a target substance, an analysis kit for a template nucleic acid or a target substance, and an analyzer for a template nucleic acid or a target substance, which are excellent in accuracy. The method for analyzing a template nucleic acid of the present invention includes the steps of fractionating a sample containing a template nucleic acid into a plurality of template nucleic acid fractions; amplifying a target sequence and its complementary sequence in the template nucleic acid with respect to each of the plurality of template nucleic acid fractions in the presence of a nucleic acid amplification reagent; detecting generation or quenching of a signal that shows an amplification of the target sequence or the complementary sequence with respect to each of the plurality of template nucleic acid fractions after the amplification step; and discriminating a template nucleic acid fraction in which the generation or quenching of a signal that shows the amplification has been detected among the plurality of template nucleic acid fractions as an amplified fraction in which the target sequence or the complementary sequence has been amplified, wherein the nucleic acid amplification reagent contains a primer set that amplifies the target sequence and the complementary sequence and a signal generating substance that generates or quenches a signal in response to the amplification, and the signal generating substance generates a signal in a state where it is bound sequence-dependently and quenches a signal in a state where it is not bound or quenches a signal in a state where it is bound sequence-dependently and generates a signal in a state where it is not bound, and generation and quenching of a signal are reversible.

2. 发明公开

EP2873731A1 NUCLEIC ACID PROBE, METHOD FOR DESIGNING NUCLEIC ACID PROBE, AND METHOD FOR DETECTING TARGET SEQUENCE 有权
标题翻译： NUCLEIC探头，方法对于核酸探针用于检测的靶序列的设计和方法
公开(公告)号：EP2873731A1
公开(公告)日：2015-05-20
申请号：EP13820349.2
申请日：2013-07-12
申请人： Kabushiki Kaisha Dnaform
发明人： HAYASHIZAKI, Yoshihide , HANAMI, Takeshi , SOMA, Takahiro , KIMURA, Yasumasa , KANAMORI, Hajime , MITANI, Yasumasa
IPC分类号： C12N15/09 , C12Q1/68
CPC分类号： C12Q1/6818 , C12N2330/30 , C12Q1/6827 , C12Q2525/117 , C12Q2535/131 , C12Q2537/101 , C12Q2563/107 , C12Q2565/1015 , C12Q2565/107
摘要： The present invention provides a nucleic acid probe that can achieve high detection sensitivity and high specificity in mutation detection, mismatch detection, etc. by the PCR method, a method for designing such a nucleic acid probe, and a method for detecting a target sequence. The nucleic acid probe includes a nucleic acid molecule, and the nucleic acid molecule includes a plurality of fluorescent dye moieties that exhibit an excitonic effect. At least two of the fluorescent dye moieties that exhibit an excitonic effect are bound to the same base or two adjacent bases in the nucleic acid molecule with each fluorescent dye moiety being bound via a linker (a linking atom or a linking atomic group). The extension-side end of the nucleic acid molecule is chemically modified, thereby preventing an extension reaction of the nucleic acid molecule.

3. 发明公开

EP2210941A1 ISOTHERMAL AMPLIFICATION METHOD AND DNA POLYMERASE USED IN THE SAME 审中-公开
标题翻译：同位素多聚核苷酸多聚酶
公开(公告)号：EP2210941A1
公开(公告)日：2010-07-28
申请号：EP08842183.9
申请日：2008-10-24
申请人： Riken , Kabushiki Kaisha Dnaform
发明人： HAYASHIZAKI, Yoshide , ITOH, Masayoshi , LEZHAVA, Alexander , BENNO, Yoshimi , MITANI, Yasumasa , KANAMORI, Hajime
IPC分类号： C12N15/09 , C12N9/12 , C12Q1/68
CPC分类号： C12Q1/6846 , C12N9/1252 , C12P19/34 , C12Q2525/155 , C12Q2521/107 , C12Q2527/101 , C12Q2531/107
摘要： A DNA polymerase suitable for specific isothermal amplification methods and an isothermal amplification method using the DNA polymerase are provided. In the presence of a DNA polymerase including a protein described in the following item (a) or (b), an amplification reaction of a target nucleic acid sequence in a nucleic acid sample is carried out isothermally using a first primer shown in the following (X). By using the DNA polymerase, it becomes possible to carry out the amplification reaction using the primer within a shorter time than ever before.
(a) a protein having an amino acid sequence represented by SEQ ID NO. 23
(b) a protein having an amino acid sequence represented by SEQ ID NO. 25

(X) a primer that contains, in a 3' end portion, a sequence (Ac') that hybridizes to a sequence (A) of a 3' end portion of the target nucleic acid sequence and also contains, on a 5' side of the sequence (Ac'), a sequence (B') that hybridizes to a complementary sequence (Bc) to a sequence (B) present on a 5' side with respect to the sequence (A) in the target nucleic acid sequence
摘要翻译：提供了适用于特定等温扩增方法的DNA聚合酶和使用DNA聚合酶的等温扩增方法。在包含下述（a）或（b）所述的蛋白质的DNA聚合酶的存在下，使用下述的第一引物等温进行靶核酸序列的扩增反应 X）。通过使用DNA聚合酶，可以在比以往更短的时间内使用引物进行扩增反应。（a）具有SEQ ID NO：1所示的氨基酸序列的蛋白质。 23（b）具有SEQ ID NO：1所示的氨基酸序列的蛋白质。 25（X）引物，其在3'末端含有与目标核酸序列的3'端部分的序列（A）杂交的序列（Ac'），并且在5' 序列（Ac'）的一侧，与互补序列（Bc）与靶核酸序列中的序列（A）存在于5'侧的序列（B）杂交的序列（B'）

4. 发明授权

EP0987335B1 Method for DNA-fixation and a DNA support 有权
标题翻译：对于DNA固定和DNS支持方法
公开(公告)号：EP0987335B1
公开(公告)日：2006-01-11
申请号：EP99117273.5
申请日：1999-09-02
申请人： THE INSTITUTE OF PHYSICAL & CHEMICAL RESEARCH , Kabushiki Kaisha Dnaform
发明人： Hayashizaki, Yoshihide
IPC分类号： C12Q1/68
CPC分类号： C12Q1/6834 , Y10S435/912

5. 发明公开

EP1564301A1 METHOD OF AMPLIFYING NUCLEIC ACID 审中-公开
标题翻译： VERFAHRENFÜRNUKLEINSÄUREAMPLIFATION
公开(公告)号：EP1564301A1
公开(公告)日：2005-08-17
申请号：EP03769966.7
申请日：2003-10-29
申请人： RIKEN , Kabushiki Kaisha Dnaform , WAKUNAGA PHARMACEUTICAL CO., LTD.
发明人： Mitani, Yasumasa, Wakunaga Pharmaceutical Co, Ltd. , Yamane, Akio, Wakunaga Pharmaceutical Co., Ltd. , Shibata, Yuko, c/o Kabushiki Kaisha Dnaform , HayashizkiI, Yoshihide
IPC分类号： C12Q1/68 , C12N15/09 , G01N33/53 , G01N33/566
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'端部分中的序列（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äX-（Y-Y'）ü/ 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表示所述目标核酸序列中的序列（A）和（B），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）。

7. 发明授权

EP3124622B1 FLUORESCENT LABELED SINGLE-STRANDED NUCLEIC ACID AND USE THEREOF 有权转让
公开(公告)号：EP3124622B1
公开(公告)日：2019-11-27
申请号：EP15773110.0
申请日：2015-03-27
申请人： Kabushiki Kaisha DNAFORM
发明人： HANAMI Takeshi , HAYASHIZAKI Yoshihide , SOMA Takahiro , KIMURA Yasumasa
IPC分类号： C12Q1/68 , C12N15/09 , G01N33/542

8. 发明授权

EP2891714B1 METHOD FOR ANALYZING TARGET NUCLEIC ACID, KIT, AND ANALYZER 有权
公开(公告)号：EP2891714B1
公开(公告)日：2018-07-11
申请号：EP13832285.4
申请日：2013-08-29
申请人： Kabushiki Kaisha Dnaform
发明人： HAYASHIZAKI Yoshihide , ITOH Masayoshi , ARAKAWA Takahiro , USUI Kengo , UEMURA Sotaro , MITANI Yasumasa
IPC分类号： C12Q1/68
CPC分类号： C12Q1/6876 , C09B23/04 , C09B23/06 , C12Q1/682 , C12Q1/6834 , C12Q1/6853 , C12Q2600/158 , C12Q2525/301 , C12Q2565/1015 , C12Q2565/107 , C12Q2565/543
摘要： The present invention is to provide a method for analyzing a target nucleic acid, by which the target nucleic acid can be analyzed rapidly and easily. In order to achieve the above object, the present invention provides a method for analyzing a target nucleic acid in a sample, including the step of analyzing the target nucleic acid in the sample by bringing the sample into contact with a label and with a primer or probe that can hybridize to the target nucleic acid. The primer or probe is immobilized on a solid phase. The label does not emit light when the primer or probe does not hybridize to the target nucleic acid, whereas the label emits light when the primer or probe has hybridized to the target nucleic acid. The analysis is carried out by detecting the light emitted from the label.

9. 发明授权

EP2894158B1 COMPOUND, NUCLEIC ACID, LABELING SUBSTANCE, AND DETECTION METHOD 有权
公开(公告)号：EP2894158B1
公开(公告)日：2018-06-20
申请号：EP13834990.7
申请日：2013-09-03
申请人： Kabushiki Kaisha DNAFORM
发明人： HAYASHIZAKI, Yoshihide , SOMA, Takahiro , HANAMI, Takeshi , KANAMORI, Hajime , BABA, Masaru
IPC分类号： C07H19/073 , C07H19/067 , C07H19/10 , C07H21/00 , C07H21/04
CPC分类号： C07H19/06 , C07H19/073 , C07H21/04 , C09B23/04 , C09B23/06 , C12Q1/6876 , Y02P20/55
摘要： The present invention provides a compound represented by the following chemical formula (I); a tautomer or stereoisomer of the compound; or a salt of the compound, the tautomer, or the stereoisomer. In the chemical formula (I), R 1 and R 2 are each a Group 1 element or a protecting group of an amino group and may be identical to or different from each other, or alternatively, R 1 and R 2 together may form a protecting group of an amino group. R 3 is a Group 1 element or a protecting group of a hydroxy group. R 4 is a Group 1 element or -PR 5 R 6 R 7 R 8 (R 5 , R 6 , R 7 , and R 8 are each a Group 1 element, a lone electron pair, a Group 16 element, a Group 17 element, or a protecting group of a phosphorus atom, and may be identical to or different from each other). J is a hydrogen atom or an arbitrary atomic group, A is a hydrogen atom, a hydroxy group, an alkyl group, an aralkyl group, an alkoxy group, an electron-withdrawing group, a silylene group, or a sulfide group, or alternatively, J and A together may form a linker. L is a single bond or a linker (a linking atom or a linking atomic group), the main chain length (the number of main chain atoms) of the linker is arbitrary, L may or may not contain each of C, N, O, S, P, and Si in the main chain, and may or may not contain each of a single bond, a double bond, a triple bond, an amide bond, an ester bond, a disulfide bond, an imino group, an ether bond, a thioether bond, and a thioester bond in the main chain. Z is an atomic group that can form a peptide bond with a labeling compound, or is an atom or atomic group including a label.

10. 发明公开

EP3266869A1 METHOD FOR ANALYZING TEMPLATE NUCLEIC ACID 审中-公开
标题翻译：分析模板核酸的方法
公开(公告)号：EP3266869A1
公开(公告)日：2018-01-10
申请号：EP16758764.1
申请日：2016-02-18
申请人： Kabushiki Kaisha DNAFORM
发明人： TSUJIMARU, Koichiro
IPC分类号： C12N15/09 , C12M1/00 , C12Q1/68 , G01N21/78
CPC分类号： C12Q1/6827 , C12M1/00 , C12N15/09 , C12Q1/6813 , C12Q1/6818 , G01N21/78 , C12Q2535/131 , C12Q2563/107 , C12Q2565/107 , C12Q2521/307 , C12Q2525/301
摘要： The present invention provides a method for accurately analyzing a template nucleic acid. The method is a method for analyzing a template nucleic acid including: an amplification step of amplifying a target sequence in the template nucleic acid and a complementary sequence to the target sequence; and a detection step of detecting association between a probe and an amplification product obtained in the amplification step or dissociation of an assembly between the probe and the amplification product with a temperature change. A primer set is used in the amplification step, and the primer set is a combination of a first primer for a polymerase that performs priming at the 3' end of a primer to synthesize the target sequence and a second primer for the polymerase to synthesize the complementary strand. Between the first primer and the second primer, a primer that initiates synthesis of a sequence to be hybridized with the probe is a turnback primer including a primer region and a probe region being directly or indirectly linked to the 5' end of the primer region, and the other primer is any primer having an ability of causing the polymerase to perform priming at the 3' end. In the amplification step, the turnback primer extends the target sequence or the complementary sequence from the primer region. In the detection step, the turnback primer also serves as the probe, and association between the probe region and the target sequence or the complementary sequence, occurred by turning the probe region of the turnback primer in a molecule of the amplification product back toward the extended target sequence or the extended complementary sequence, is detected
摘要翻译：本发明提供了用于准确分析模板核酸的方法。该方法是用于分析模板核酸的方法，其包括：扩增模板核酸中的靶序列和与靶序列互补的序列的扩增步骤; 以及检测步骤，检测在扩增步骤中获得的探针和扩增产物之间的关联或者探针和扩增产物之间的组件与温度变化的解离。在扩增步骤中使用引物组，并且引物组是用于在引物的3'末端进行引发以合成靶序列的聚合酶的第一引物和用于聚合酶合成互补链。在第一引物和第二引物之间，引发与探针杂交的序列的合成的引物是包括引物区域和与引物区域的5'末端直接或间接连接的探针区域的折返引物，而另一个引物是具有引起聚合酶在3'末端进行引发的能力的任何引物。在扩增步骤中，折返引物将来自引物区域的靶序列或互补序列延伸。在检测步骤中，回折引物也用作探针，并且通过将扩增产物分子中的折回引物的探针区域向扩展方向返回而发生探针区域与靶序列或互补序列之间的结合目标序列或扩展的互补序列被检测到

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