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1. 发明申请

US20090311770A1 Method of collecting microorganisms using fine particles, method of collecting nucleic acids using fine particles, and kits for use in the these methods 审中-公开
标题翻译：使用细颗粒收集微生物的方法，使用细颗粒收集核酸的方法，以及用于这些方法的试剂盒
公开(公告)号：US20090311770A1
公开(公告)日：2009-12-17
申请号：US11919475
申请日：2006-05-19
申请人： Satoshi Hashiguchi , Mitsuharu Hirai , Toshiya Hosomi
发明人： Satoshi Hashiguchi , Mitsuharu Hirai , Toshiya Hosomi
IPC分类号： C12N1/02 , C12N7/02 , C12N1/20 , C12N5/06
CPC分类号： C12N7/00 , C12N1/02 , C12N15/1013 , C12N2730/10151 , C12N2740/16051 , C12N2770/24251 , C12Q1/24
摘要： The present invention provides a method of collecting microorganisms and a method of collecting nucleic acids, which both can be carried out easily and can achieve a high collection rate. A method of collecting microorganisms according to the present invention includes a microorganism adsorption step of bringing a sample into contact with fine particles so as to cause microorganisms contained in the sample to be adsorbed onto the fine particles. In this method, the fine particles have a particle diameter of 6 μm or less and a specific surface area of 50 m2/g or less. Furthermore, a method of collecting nucleic acids according to the present invention includes: a microorganism adsorption step of causing microorganisms to be adsorbed onto fine particles; and a nucleic acid elution step of eluting nucleic acids from the microorganisms that have been adsorbed onto the fine particles. The microorganism adsorption step in this method is the microorganism collection method of the present invention. According to the collection methods of the present invention, microorganisms and nucleic acids can be collected easily and efficiently. Preferably, the fine particles are magnetic silica particles.
摘要翻译：本发明提供收集微生物的方法和收集核酸的方法，这两者都可以容易地进行并且可以实现高收集率。根据本发明的收集微生物的方法包括微生物吸附步骤，使样品与细颗粒接触，以使样品中所含的微生物吸附到微粒上。在该方法中，微粒的粒径为6μm以下，比表面积为50m 2 / g以下。此外，根据本发明的收集核酸的方法包括：使微生物吸附在细颗粒上的微生物吸附步骤; 以及从已经吸附到微粒上的微生物中洗脱核酸的核酸洗脱步骤。该方法中的微生物吸附步骤是本发明的微生物收集方法。根据本发明的收集方法，可以容易且有效地收集微生物和核酸。优选地，微粒是磁性二氧化硅颗粒。

2. 发明申请

US20130017543A1 Method and Kit for Amplifying and Detecting Polynucleotide 有权
标题翻译：用于扩增和检测多核苷酸的方法和试剂盒
公开(公告)号：US20130017543A1
公开(公告)日：2013-01-17
申请号：US13545766
申请日：2012-07-10
申请人： Toshiya Hosomi , Mitsuharu Hirai
发明人： Toshiya Hosomi , Mitsuharu Hirai
IPC分类号： C12P19/34 , C12N9/12 , C12Q1/68
CPC分类号： C12Q1/686 , C12Q2521/101 , C12Q2521/107
摘要： Disclosed is a method of amplifying a polynucleotide, comprising: (a) mixing primers for amplifying the polynucleotide, a polymerase, nucleotide substrates and a template polynucleotide, and (b) amplifying the polynucleotide by polymerase reaction, wherein the polymerase has an amino acid sequence consisting of SEQ ID NO:1 or an amino acid sequence with at least 85% sequence identity to SEQ ID NO:1, and an amino acid residue corresponding to, or at position 651 of the amino acid sequence has been replaced with glutamic acid.
摘要翻译：公开了扩增多核苷酸的方法，包括：（a）混合用于扩增多核苷酸的引物，聚合酶，核苷酸底物和模板多核苷酸，和（b）通过聚合酶反应扩增多核苷酸，其中所述聚合酶具有氨基酸序列由SEQ ID NO：1或与SEQ ID NO：1具有至少85％序列同一性的氨基酸序列组成，氨基酸序列的氨基酸残基与氨基酸序列相应或位置651被氨基酸替代。

3. 发明授权

US09045797B2 Method and kit for amplifying and detecting polynucleotide 有权
标题翻译：用于扩增和检测多核苷酸的方法和试剂盒
公开(公告)号：US09045797B2
公开(公告)日：2015-06-02
申请号：US13545766
申请日：2012-07-10
申请人： Toshiya Hosomi , Mitsuharu Hirai
发明人： Toshiya Hosomi , Mitsuharu Hirai
IPC分类号： C12Q1/68 , C12P19/34
CPC分类号： C12Q1/686 , C12Q2521/101 , C12Q2521/107
摘要： Disclosed is a method of amplifying a polynucleotide, comprising: (a) mixing primers for amplifying the polynucleotide, a polymerase, nucleotide substrates and a template polynucleotide, and (b) amplifying the polynucleotide by polymerase reaction, wherein the polymerase has an amino acid sequence consisting of SEQ ID NO:1 or an amino acid sequence with at least 85% sequence identity to SEQ ID NO:1, and an amino acid residue corresponding to, or at position 651 of the amino acid sequence has been replaced with glutamic acid.
摘要翻译：公开了扩增多核苷酸的方法，包括：（a）混合用于扩增多核苷酸的引物，聚合酶，核苷酸底物和模板多核苷酸，和（b）通过聚合酶反应扩增多核苷酸，其中所述聚合酶具有氨基酸序列由SEQ ID NO：1或与SEQ ID NO：1具有至少85％序列同一性的氨基酸序列组成，氨基酸序列的氨基酸残基与氨基酸序列相应或位置651被氨基酸替代。

4. 发明授权

US08889359B2 Method and kit for amplifying and detecting polynucleotide using a modified Tth polymerase 有权
标题翻译：使用修饰的第T聚合酶扩增和检测多核苷酸的方法和试剂盒
公开(公告)号：US08889359B2
公开(公告)日：2014-11-18
申请号：US13545756
申请日：2012-07-10
申请人： Toshiya Hosomi , Mitsuharu Hirai
发明人： Toshiya Hosomi , Mitsuharu Hirai
IPC分类号： C12Q1/68 , C12N9/02
CPC分类号： C12Q1/6844 , C12Q1/686 , C12Q2521/101 , C12Q2521/107
摘要： Disclosed is a method of amplifying a polynucleotide, comprising: (a) mixing primers for amplifying the polynucleotide, a polymerase, nucleotide substrates and a template polynucleotide, and (b) amplifying the polynucleotide by polymerase reaction, wherein the polymerase has an amino acid sequence consisting of SEQ ID NO: 1 or an amino acid sequence with at least 85% sequence identity to SEQ ID NO: 1, and wherein an amino acid residue corresponding to position 653 of the amino acid sequence has been replaced with glutamic acid.
摘要翻译：公开了扩增多核苷酸的方法，包括：（a）混合用于扩增多核苷酸的引物，聚合酶，核苷酸底物和模板多核苷酸，和（b）通过聚合酶反应扩增多核苷酸，其中所述聚合酶具有氨基酸序列由SEQ ID NO：1或与SEQ ID NO：1具有至少85％序列同一性的氨基酸序列组成，其中对应于氨基酸序列的位置653的氨基酸残基已被谷氨酸替代。

5. 发明申请

US20090113980A1 METHOD OF VERIFYING PERFORMANCE OF OPTICAL DETECTION APPARATUS AND STANDARD REAGENT USED THEREFOR 有权
标题翻译：验证光学检测装置性能的方法及其使用的标准试剂
公开(公告)号：US20090113980A1
公开(公告)日：2009-05-07
申请号：US12300510
申请日：2007-12-25
申请人： Mitsuharu Hirai , Toshiya Hosomi , Yuki Yoshinaga
发明人： Mitsuharu Hirai , Toshiya Hosomi , Yuki Yoshinaga
IPC分类号： G01N37/00 , C07H21/00 , G01J1/10
CPC分类号： G01N21/274 , C12Q1/6816 , C12Q1/6848 , C12Q2561/113 , C12Q2545/113 , C12Q2527/107
摘要： A method is provided in which with respect to an optical detection apparatus including an optical detection unit and a temperature control unit, whether optical signal detection and temperature control are performed accurately, i.e. the performance thereof, can be verified simply with high reliability. With respect to an optical detection apparatus including an optical detection unit for detecting an optical signal of a sample and a temperature control unit for controlling temperature of the sample, the optical signal detection performance and temperature control performance are verified by the following method. First, a standard sample containing a nucleic acid sequence and a strand complementary thereto that have a known optical signal intensity and Tm value is provided, the temperature of the standard sample is increased or decreased with the temperature control unit, and optical signal intensity of the standard sample is measured with the detection unit. On the other hand, the melting temperature of the standard sample is determined from a change in the optical signal intensity accompanying a change in the temperature. The measured optical signal intensity and melting temperature of the standard sample are compared to the known optical signal intensity and melting temperature of the standard sample, respectively, and thereby it is verified whether the optical signal detection performance of the detection unit and the temperature control performance of the temperature control unit are accurate.
摘要翻译：提供一种方法，其中对于包括光学检测单元和温度控制单元的光学检测装置，可以简单地以高可靠性来验证光学信号检测和温度控制是否被精确地执行，即其性能。对于包括用于检测样品的光学信号的光学检测单元和用于控制样品的温度的温度控制单元的光学检测设备，通过以下方法验证光学信号检测性能和温度控制性能。首先，提供含有具有已知光信号强度和Tm值的核酸序列和与其互补的链的标准样品，标准样品的温度随着温度控制单元而增加或减小，光信号强度用检测单元测量标准样品。另一方面，标准样品的熔解温度由伴随温度变化的光信号强度的变化确定。将标准样品的测量光信号强度和熔融温度分别与标准样品的已知光信号强度和熔融温度进行比较，从而验证检测单元的光信号检测性能和温度控制性能的温度控制单元是准确的。

6. 发明授权

US09121839B2 Analyzing system, analyzing apparatus, container, analyzing method, program, and recording medium 有权
标题翻译：分析系统，分析仪器，容器，分析方法，程序和记录介质
公开(公告)号：US09121839B2
公开(公告)日：2015-09-01
申请号：US13145774
申请日：2010-01-21
申请人： Mitsuharu Hirai , Satoshi Majima , Toshiya Hosomi
发明人： Mitsuharu Hirai , Satoshi Majima , Toshiya Hosomi
IPC分类号： G01N21/75 , G01N35/00 , B01L3/00 , G01N35/02
CPC分类号： G01N35/00732 , B01L3/5085 , B01L3/527 , B01L3/545 , B01L2200/04 , B01L2300/02 , B01L2300/021 , B01L2300/022 , B01L2300/023 , B01L2300/024 , B01L2300/0829 , G01N21/75 , G01N35/026 , G01N2035/00752
摘要： An analyzing system that enables further expansion of analysis items and automation of analysis. In the analyzing system for performing an analysis using container 1 and an analyzing apparatus, container 1 is a dedicated container previously containing a reagent for a specific analysis item or an expansion container to which a user can freely set an analysis item.
摘要翻译：一个分析系统，可进一步扩展分析项目和分析自动化。在用于使用容器1和分析装置进行分析的分析系统中，容器1是预先包含用于特定分析项目的试剂或膨胀容器的专用容器，用户可以自由地设定分析项目。

7. 发明授权

US09115391B2 Method of detecting a polymorphism at a polymorphism site 有权
标题翻译：检测多态性位点多态性的方法
公开(公告)号：US09115391B2
公开(公告)日：2015-08-25
申请号：US13002194
申请日：2009-07-02
申请人： Mitsuharu Hirai , Toshiya Hosomi , Aki Iguchi
发明人： Mitsuharu Hirai , Toshiya Hosomi , Aki Iguchi
IPC分类号： C12Q1/68 , C12P19/34
CPC分类号： C12Q1/6827 , C12Q1/6858 , C12Q2545/101 , C12Q2535/125 , C12Q2527/107
摘要： The present invention provides a method for detecting a mutation capable of detecting a mutation with high sensitivity and high reliability in one reaction system. Using primers (Xmt) and (Xwt), a target nucleic acid sequence whose objective base to be detected is a mutant-type is amplified with amplification efficiency higher than a target nucleic acid sequence whose objective base to be detected is a normal-type. The (Xmt) is a primer that is complementary to a region including a mutant-type base in the template nucleic acid and has a base complementary to a mutant-type base at a 3′ region, and the (Xwt) is a primer that is complementary to a region including a normal-type base in the template nucleic acid and has a base complementary to a normal-type base at a 3′ region. It is preferable that amplification efficiency by the (Xmt) with reference to a mutant-type template nucleic acid is higher than that by the (Xwt) with reference to a normal-type template nucleic acid. Then, a signal value that shows a molten state of a hybridization product between the thus obtained amplification product and the probe is measured, and the presence or absence of the mutation of the objective base site is determined from a change in the signal value accompanying a change in the temperature.
摘要翻译：本发明提供一种用于检测在一个反应体系中能够检测高灵敏度和高可靠性的突变的突变的方法。使用引物（Xmt）和（Xwt），以待检测的目的基因为突变型的靶核酸序列的扩增效率高于目标基因为正常型的靶核酸序列。（Xmt）是与模板核酸中包含突变型碱基的区域互补的引物，并且在3'区域具有与突变型碱基互补的碱基，并且（Xwt）是引物，与包含模板核酸中的正常型碱基的区域互补，并且在3'区域具有与正常型碱基互补的碱基。相对于突变型模板核酸，优选（Xmt）的扩增效率高于参照正常型模板核酸的（Xwt）的扩增效率。然后，测定显示由此获得的扩增产物和探针之间的杂交产物的熔融状态的信号值，并且根据伴随的信号值的变化确定目标碱基位点的突变的存在或不存在温度变化。

8. 发明申请

US20130017542A1 Method and Kit for Amplifying and Detecting Polynucleotide 有权
标题翻译：用于扩增和检测多核苷酸的方法和试剂盒
公开(公告)号：US20130017542A1
公开(公告)日：2013-01-17
申请号：US13545756
申请日：2012-07-10
申请人： Toshiya Hosomi , Mitsuharu Hirai
发明人： Toshiya Hosomi , Mitsuharu Hirai
IPC分类号： C12P19/34 , G01N25/04 , C12N9/12 , C12Q1/68
CPC分类号： C12Q1/6844 , C12Q1/686 , C12Q2521/101 , C12Q2521/107
摘要： Disclosed is a method of amplifying a polynucleotide, comprising: (a) mixing primers for amplifying the polynucleotide, a polymerase, nucleotide substrates and a template polynucleotide, and (b) amplifying the polynucleotide by polymerase reaction, wherein the polymerase has an amino acid sequence consisting of SEQ ID NO:1 or an amino acid sequence with at least 85% sequence identity to SEQ ID NO:1, and wherein an amino acid residue corresponding to position 653 of the amino acid sequence has been replaced with glutamic acid.
摘要翻译：公开了扩增多核苷酸的方法，包括：（a）混合用于扩增多核苷酸的引物，聚合酶，核苷酸底物和模板多核苷酸，和（b）通过聚合酶反应扩增多核苷酸，其中所述聚合酶具有氨基酸序列由SEQ ID NO：1或与SEQ ID NO：1具有至少85％序列同一性的氨基酸序列组成，其中对应于氨基酸序列的位置653的氨基酸残基已被谷氨酸替代。

9. 发明申请

US20120021419A1 Analyzing System, Analyzing Apparatus, Container, Analyzing Method, Program, and Recording Medium 有权
标题翻译：分析系统，分析仪器，容器，分析方法，程序和记录介质
公开(公告)号：US20120021419A1
公开(公告)日：2012-01-26
申请号：US13145774
申请日：2010-01-21
申请人： Mitsuharu Hirai , Satoshi Majima , Toshiya Hosomi
发明人： Mitsuharu Hirai , Satoshi Majima , Toshiya Hosomi
IPC分类号： C12Q1/68 , C12M1/34
CPC分类号： G01N35/00732 , B01L3/5085 , B01L3/527 , B01L3/545 , B01L2200/04 , B01L2300/02 , B01L2300/021 , B01L2300/022 , B01L2300/023 , B01L2300/024 , B01L2300/0829 , G01N21/75 , G01N35/026 , G01N2035/00752
摘要： An analyzing system that enables further expansion of analysis items and automation of analysis. In the analyzing system for performing an analysis using container 1 and an analyzing apparatus, container 1 is a dedicated container previously containing a reagent for a specific analysis item or an expansion container to which a user can freely set an analysis item.
摘要翻译：一个分析系统，可进一步扩展分析项目和分析自动化。在用于使用容器1和分析装置进行分析的分析系统中，容器1是预先包含用于特定分析项目的试剂或膨胀容器的专用容器，用户可以自由地设定分析项目。

10. 发明申请

US20100273173A1 METHOD FOR AMPLIFYING TARGET NUCLEIC ACID SEQUENCE AND PROBE USED FOR THE SAME 审中-公开
标题翻译：用于放大目标核酸序列的方法和用于其的探针
公开(公告)号：US20100273173A1
公开(公告)日：2010-10-28
申请号：US12810266
申请日：2008-12-25
申请人： Mitsuharu Hirai , Satoshi Majima , Toshiya Hosomi
发明人： Mitsuharu Hirai , Satoshi Majima , Toshiya Hosomi
IPC分类号： C12Q1/68 , C12P19/34 , C07H21/00
CPC分类号： C12Q1/6827 , C12Q1/6844 , C12Q2527/107
摘要： The present invention provides a method for amplifying a target sequence while suppressing inhibition of an amplification reaction in nucleic acid amplification in the presence of the probe. At the time of amplifying the target sequence, as the probe caused to coexist in amplification of the target sequence, a probe having a base sequence in which a melting temperature of the double-stranded nucleic acid is equal to or lower than a reaction temperature of the elongation reaction is used. In the presence of such a probe, for example, annealing of a primer and an elongation reaction from the primer are hardly inhibited by the presence of the probe so that amplification of the target sequence can be conducted sufficiently. Therefore, when a polymorphism of a target site in the target sequence is analyzed by a Tm analysis or the like, high reliability can be achieved.
摘要翻译：本发明提供了在存在探针的同时抑制核酸扩增中扩增反应的抑制的同时扩增靶序列的方法。在扩增靶序列时，作为探针在扩增靶序列时共存的探针，具有碱基序列的探针，其中双链核酸的解链温度等于或低于使用伸长反应。在这样的探针的存在下，例如引物的退火和引物的延伸反应几乎不被探针的存在抑制，从而可以充分进行目标序列的扩增。因此，当通过Tm分析等分析目标序列中的靶位点的多态性时，可以实现高可靠性。

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