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1. 发明申请

US20060210988A1 Method for isolating nucleic acid and, for nucleic acid isolation, kit and apparatus 审中-公开
标题翻译：用于分离核酸的方法，用于核酸分离的试剂盒和装置
公开(公告)号：US20060210988A1
公开(公告)日：2006-09-21
申请号：US10553376
申请日：2004-04-22
申请人： Ken Inose , Satoshi Hashiguchi , Yuji Izumizawa
发明人： Ken Inose , Satoshi Hashiguchi , Yuji Izumizawa
IPC分类号： C12Q1/68 , C12N1/08 , C12M1/34
CPC分类号： C12N15/1003
摘要： Nucleic acids suitable for PCR amplification are isolated from a sample easily and rapidly by dissolving the sample in a buffer containing surfactant and salt, heating the obtained solution, subjecting the heated solution to gel filtration; and collecting a fraction containing nucleic acids.
摘要翻译：通过将样品溶解在含有表面活性剂和盐的缓冲液中，将所得溶液加热，使加热溶液进行凝胶过滤，从样品中分离出适合于PCR扩增的核酸; 并收集含有核酸的级分。

2. 发明申请

US20080035482A1 Method Of Concentrating And Purifying Nucleic Acid And Apparatus Therefor 审中-公开
标题翻译：浓缩和纯化核酸的方法及其设备
公开(公告)号：US20080035482A1
公开(公告)日：2008-02-14
申请号：US10578770
申请日：2004-11-09
申请人： Satoshi Hashiguchi , Ken Inose
发明人： Satoshi Hashiguchi , Ken Inose
IPC分类号： C12N15/00 , C12M1/00 , C12Q1/68 , G01N27/447
CPC分类号： C12N15/1017 , B01D57/02 , B01D61/145 , B01D61/42 , B01D61/425 , B01D61/46 , B01L3/502753 , C12N15/101 , G01N27/44747
摘要： The conventional method of purifying and concentrating nucleic acids, because of dangerous chemicals, requires elaborate chemical equipment to result in restriction of environment available. Further, time-consuming operation is inevitable and high-speed centrifugation, etc. are needed to cause automation to be difficult and to cause high purification degree to be unattainable. Still further, in the purification method using a column/filter, application of dusty samples tends to invite clogging to lead to a drop of purification efficiency, and centrifugation or suction operation is needed to cause automation to be difficult. In this invention, surfactants (3,4) are adsorbed on impurity (2) contained in a sample, so that the impurity (2) conducts behavior different from that of nucleic acid (1) to thereby attain separation of the impurity (2) from the nucleic acid (1). Impurity (2) other than nucleic acid (1) is energized with cationic surfactant (4) and nonionic surfactant (3) and placed in an electric field to thereby effect separation and purification of the nucleic acid (1) for an analyte containing the impurity (2). Thus, the nucleic acid (1) is brought into the state of being concentrated or easily concentrated.
摘要翻译：由于危险化学品，纯化和浓缩核酸的常规方法需要精心设计的化学设备来限制可用的环境。此外，耗时的操作是不可避免的，需要高速离心等来引起自动化困难，并且导致高的净化程度是不可实现的。此外，在使用柱/过滤器的净化方法中，灰尘样品的施用往往会引起堵塞，导致净化效率降低，需要离心或抽吸操作以使自动化变得困难。在本发明中，表面活性剂（3,4）被吸附在样品中所含的杂质（2）上，使得杂质（2）的导电性质与核酸（1）不同，从而使杂质（2）分离，来自核酸（1）。核酸（1）以外的杂质（2）用阳离子表面活性剂（4）和非离子表面活性剂（3）通电，并放置在电场中，从而对含有杂质的分析物（1）进行分离和纯化（2）。因此，使核酸（1）进入浓缩或容易浓缩的状态。

3. 发明申请

US20060188881A1 Dna amplification method and kit therefor 失效
标题翻译： Dna扩增方法及其试剂盒
公开(公告)号：US20060188881A1
公开(公告)日：2006-08-24
申请号：US10547354
申请日：2004-03-05
申请人： Satoshi Hashiguchi , Ken Inose
发明人： Satoshi Hashiguchi , Ken Inose
IPC分类号： C12Q1/68 , C12P19/34
CPC分类号： C12Q1/6853 , C12Q1/686 , C12Q2525/155
摘要： In a method for amplifying a DNA, comprising performing PCR using a sense primer and an antisense primer, PCR is performed in the presence of an additional sense primer comprising the sense primer and an oligodeoxyribonucleotide having a first additional sequence and ligated to the 5′ end of the sense primer and an additional antisense primer comprising the antisense primer and an oligodeoxyribonucleotide having a second additional sequence complementary to the first additional sequence and ligated to the 5′ end of the antisense primer; a Tm value of the additional sequences is lower than Tm values of the sense primer and the antisense primer; and annealing temperature in PCR is initially set to be a temperature at which the additional sequences do not anneal and changed in the course of PCR to a temperature at which the additional sequences anneal to each other.
摘要翻译：在扩增DNA的方法中，包括使用有义引物和反义引物进行PCR，在包含有义引物和具有第一附加序列的寡脱氧核糖核苷酸的另外的有义引物的存在下进行PCR，并连接到5'末端的正义引物和另外的反义引物，其包含反义引物和具有与第一附加序列互补的第二附加序列并连接到反义引物的5'末端的寡脱氧核糖核苷酸; 附加序列的Tm值低于有义引物和反义引物的Tm值; 并且PCR中的退火温度最初设定为附加序列在PCR过程中不退火并且在附加序列彼此退火的温度下的温度。

4. 发明申请

US20060134773A1 Device for pretreating specimen 审中-公开
标题翻译：预处理试样的装置
公开(公告)号：US20060134773A1
公开(公告)日：2006-06-22
申请号：US10536827
申请日：2003-11-27
申请人： Ken Inose , Shinya Nakajima , Satoshi Hashiguchi , Jun Murakami
发明人： Ken Inose , Shinya Nakajima , Satoshi Hashiguchi , Jun Murakami
IPC分类号： C12M1/34
CPC分类号： B01L3/5027 , B01L2200/0631 , B01L2300/0681 , B01L2300/0803 , B01L2300/0816 , B01L2300/087 , B01L2300/0874 , B01L2300/0877 , B01L2300/1827 , B01L2400/0406 , B01L2400/0421 , B01L2400/0487 , B01L2400/0644
摘要： Due to the recent decipherment of the human genome, various relations between life processes and genes have been analyzed. Consequently, medical importance shifts from pathology to etiology, and from medical treatment to prevention. Hereupon, the gene test technology becomes an important basis. A pretreatment device for pretreating a specimen comprises a specimen introducing portion, a holding portion, a wash storage, an elute storage, and a discharging portion in order to prevent a loss of sensitivity during a pretreatment process of the nucleic acid test, to easily automate the pretreatment process, and to decrease the cost thereof, thereby serving as a familiar system for the gene test.
摘要翻译：由于人类基因组最近的解密，人们分析了生命过程和基因之间的各种关系。因此，医学重要性从病理学转变为病因学，从医疗转向预防。因此，基因检测技术成为重要的依据。用于预处理样品的预处理装置包括样品导入部分，保持部分，洗涤储存器，洗脱储存器和排出部分，以便防止在核酸测试的预处理过程中的敏感性损失，以便容易地自动化预处理过程，并降低成本，从而作为基因测试的熟悉系统。

5. 发明申请

US20090000949A1 Method And Apparatus Of Concentration And Purification Of Nucleic Acid 审中-公开
标题翻译：核酸浓缩和纯化的方法和装置
公开(公告)号：US20090000949A1
公开(公告)日：2009-01-01
申请号：US12182004
申请日：2008-07-29
申请人： Mitsuharu Hirai , Jun Murakami , Satoshi Hashiguchi , Ken Inose
发明人： Mitsuharu Hirai , Jun Murakami , Satoshi Hashiguchi , Ken Inose
IPC分类号： G01N27/447
CPC分类号： C12N15/1017 , B01D61/425 , B01D61/46 , C07H21/00 , C12N15/101 , G01N27/44747
摘要： In a method for concentration and purification method of nucleic acids using electrophoresis, cationic surfactant is added to a sample containing nucleic acids so as to adjust electric charge of impurities in the sample, and then the sample is placed in an electric field for electrophoresis so as to concentrate and purify the nucleic acids. The cationic surfactant (4) adsorbs substance other than the nucleic acids so as to adjust the electric charge of the substance, and the adsorption of the cationic surfactant (4) is adjusted by the added amount of nonionic surfactant (3). Alternatively, nucleic acids are contacted with a separation medium (108), and then recovered by a filter (104) whose cross sectional area is decreased in the direction of migration.
摘要翻译：在使用电泳的核酸的浓缩和纯化方法的方法中，将阳离子表面活性剂加入到含有核酸的样品中，以调节样品中杂质的电荷，然后将样品置于电场中进行电泳，以便浓缩和纯化核酸。阳离子表面活性剂（4）吸附核酸以外的物质，以调节物质的电荷，并通过非离子表面活性剂（3）的添加量调节阳离子表面活性剂（4）的吸附。或者，核酸与分离介质（108）接触，然后通过其横截面积沿迁移方向减小的过滤器（104）回收。

6. 发明授权

US07365187B2 DNA amplification method and kit therefor 失效
标题翻译： DNA扩增方法及试剂盒
公开(公告)号：US07365187B2
公开(公告)日：2008-04-29
申请号：US10547354
申请日：2004-03-05
申请人： Satoshi Hashiguchi , Ken Inose
发明人： Satoshi Hashiguchi , Ken Inose
IPC分类号： C07H21/04 , C12Q1/68 , C12P19/34
CPC分类号： C12Q1/6853 , C12Q1/686 , C12Q2525/155
摘要： In a method for amplifying a DNA, comprising performing PCR using a sense primer and an antisense primer, PCR is performed in the presence of an additional sense primer comprising the sense primer and an oligodeoxyribonucleotide having a first additional sequence and ligated to the 5′ end of the sense primer and an additional antisense primer comprising the antisense primer and an oligodeoxyribonucleotide having a second additional sequence complementary to the first additional sequence and ligated to the 5′ end of the antisense primer; a Tm value of the additional sequences is lower than Tm values of the sense primer and the antisense primer; and annealing temperature in PCR is initially set to be a temperature at which the additional sequences do not anneal and changed in the course of PCR to a temperature at which the additional sequences anneal to each other.
摘要翻译：在扩增DNA的方法中，包括使用有义引物和反义引物进行PCR，在包含有义引物和具有第一附加序列的寡脱氧核糖核苷酸的另外的有义引物的存在下进行PCR，并连接到5'末端的正义引物和另外的反义引物，其包含反义引物和具有与第一附加序列互补的第二附加序列并连接到反义引物的5'末端的寡脱氧核糖核苷酸; 附加序列的Tm值低于有义引物和反义引物的Tm值; 并且PCR中的退火温度最初设定为附加序列在PCR过程中不退火并且在附加序列彼此退火的温度下的温度。

7. 发明申请

US20060134620A1 Method and apparatus for concentration and purification of nucleic acid 审中-公开
标题翻译：核酸浓缩和纯化的方法和装置
公开(公告)号：US20060134620A1
公开(公告)日：2006-06-22
申请号：US10536822
申请日：2003-11-27
申请人： Mitsuharu Hirai , Jun Murakami , Satoshi Hashiguchi , Ken Inose
发明人： Mitsuharu Hirai , Jun Murakami , Satoshi Hashiguchi , Ken Inose
IPC分类号： C12Q1/68 , G01F1/64
CPC分类号： C12N15/1017 , B01D61/425 , B01D61/46 , C07H21/00 , C12N15/101 , G01N27/44747
摘要： In a method for concentration and purification method of nucleic acids using electrophoresis, cationic surfactant is added to a sample containing nucleic acids so as to adjust electric charge of impurities in the sample, and then the sample is placed in an electric field for electrophoresis so as to concentrate and purify the nucleic acids. The cationic surfactant (4) adsorbs substance other than the nucleic acids so as to adjust the electric charge of the substance, and the adsorption of the cationic surfactant (4) is adjusted by the added amount of nonionic surfactant (3). Alternatively, nucleic acids are contacted with a separation medium (108), and then recovered by a filter (104) whose cross sectional area is decreased in the direction of migration.
摘要翻译：在使用电泳的核酸的浓缩和纯化方法的方法中，将阳离子表面活性剂加入到含有核酸的样品中，以调节样品中杂质的电荷，然后将样品置于电场中进行电泳，以便浓缩和纯化核酸。阳离子表面活性剂（4）吸附核酸以外的物质，以调节物质的电荷，并通过非离子表面活性剂（3）的添加量调节阳离子表面活性剂（4）的吸附。或者，核酸与分离介质（108）接触，然后通过其横截面积沿迁移方向减小的过滤器（104）回收。

8. 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。

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