公开(公告)号：US09139866B2

公开(公告)日：2015-09-22

申请号：US11994823

申请日：2006-02-17

申请人： Shinichi Yoshida ,  Takashi Soejima

发明人： Shinichi Yoshida ,  Takashi Soejima

IPC分类号： C12Q1/68 ,  C07H21/04 ,  C07H21/02

CPC分类号： C12Q1/6806 ,  C12Q1/686 ,  C12Q1/6888 ,  C12Q1/689 ,  C12Q2600/158 ,  C12Q2521/519

摘要： Live cells of a microorganism in a test sample are detected by the following steps: a) the step of treating the test sample with a topoisomerase poison and/or a DNA gyrase poison, b) the step of extracting DNA from the test sample, and amplifying a target region of the extracted DNA by PCR, and c) the step of analyzing an amplification product.

摘要翻译： 通过以下步骤检测测试样品中微生物的活细胞：a）用拓扑异构酶毒物和/或DNA促旋转染酶毒物处理测试样品的步骤，b）从测试样品中提取DNA的步骤，以及 通过PCR扩增提取的DNA的靶区域，和c）分析扩增产物的步骤。

公开(公告)号：US20120277121A1

公开(公告)日：2012-11-01

申请号：US13523738

申请日：2012-06-14

申请人： Shinichi Yoshida ,  Takashi Soejima

发明人： Shinichi Yoshida ,  Takashi Soejima

IPC分类号： C12Q1/68 ,  C40B40/06

CPC分类号： C12Q1/68 ,  C12Q1/689 ,  C12Q2523/101

摘要： A kit for use in a method for detecting live cells, injured cells and dead cells of a microorganism in a test sample by a nucleic acid amplification method is disclosed. The kit includes a cross-linker capable of cross-linking DNA by irradiation with light having a wavelength of 350 nm to 700 nm, medium, and a primer (s) for amplifying a DNA target region in the microorganism by a nucleic acid amplification method. The nucleic acid amplification method may be a PCR, LAMP, SDA, LCR or DNA microarray method. A cross-linker may be included in the kit such as ethidium monoazide, ethidium diazide, psoralen, 4,5′,8-trimethyl psoralen, or 8-methoxy psoralen.

摘要翻译： 公开了一种用于通过核酸扩增方法检测测试样品中微生物的活细胞，受损细胞和死细胞的方法的试剂盒。 试剂盒包括能够通过用波长为350nm至700nm的光照射DNA和通过核酸扩增方法扩增微生物中的DNA靶标区域的引物的交联剂 。 核酸扩增方法可以是PCR，LAMP，SDA，LCR或DNA微阵列法。 试剂盒中可以包括交联剂，例如单乙叠胺，二氮嗪，补骨脂素，4,5'，8-三甲基补骨脂素或8-甲氧基补骨脂素。

公开(公告)号：US08241866B2

公开(公告)日：2012-08-14

申请号：US12832740

申请日：2010-07-08

申请人： Shinichi Yoshida ,  Takashi Soejima

发明人： Shinichi Yoshida ,  Takashi Soejima

IPC分类号： C12Q1/02 ,  C12Q1/04 ,  C12Q1/18

CPC分类号： C12Q1/04 ,  C12Q1/34 ,  C12Q1/533 ,  C12Q2304/10

摘要： A kit is disclosed for preparing a measurement sample for detecting live cells, injured cells, VNC cells and dead microorganism cells in a test sample by the following steps: a) the step of treating the test sample with an enzyme having an activity of decomposing cells other than those of the microorganism, colloidal particles of proteins or lipids existing in the test sample, b) the step of treating the test sample with a topoisomerase poison and/or a DNA gyrase poison. c) the step of treating the test sample treated in the steps a) and b) with a nuclear stain agent, and d) the step of detecting the microorganism in the test sample treated with the nuclear stain agent by flow cytometry.

摘要翻译： 公开了用于通过以下步骤制备用于检测测试样品中的活细胞，受损细胞，VNC细胞和死亡微生物细胞的测量样品的试剂盒：a）用具有分解细胞活性的酶处理测试样品的步骤 除了微生物之外，存在于测试样品中的蛋白质或脂质的胶体颗粒，b）用拓扑异构酶毒物和/或DNA回旋酶毒物处理测试样品的步骤，c）处理测试样品的步骤 在步骤a）和b）中使用核染色剂，以及d）通过流式细胞术检测用核染色剂处理的测试样品中的微生物的步骤。

公开(公告)号：US08143018B2

公开(公告)日：2012-03-27

申请号：US12832782

申请日：2010-07-08

申请人： Shinichi Yoshida ,  Takashi Soejima

发明人： Shinichi Yoshida ,  Takashi Soejima

IPC分类号： C12Q1/02 ,  C12Q1/04 ,  C12Q1/18

CPC分类号： C12Q1/04 ,  C12Q1/34 ,  C12Q1/533 ,  C12Q2304/10

摘要： According to the following steps, live cells, injured cells, VNC cells and dead cells of a microorganism in a test sample are detected by flow cytometry: a) the step of treating the test sample with an enzyme having an activity of decomposing cells other than those of the microorganism, colloidal particles of proteins or lipids existing in the test sample, b) the step of treating the test sample with a topoisomerase poison and/or a DNA gyrase poison. c) the step of treating the test sample treated in the steps a) and b) with a nuclear stain agent, and d) the step of detecting the microorganism in the test sample treated with the nuclear stain agent by flow cytometry.

摘要翻译： 根据以下步骤，通过流式细胞术检测试验样品中微生物的活细胞，受损细胞，VNC细胞和死细胞：a）用具有分解细胞以外的细胞活性的酶处理测试样品的步骤 微生物的那些，存在于测试样品中的蛋白质或脂质的胶体颗粒，b）用拓扑异构酶毒物和/或DNA回旋酶毒物处理测试样品的步骤，c）处理在测试样品中处理的测试样品的步骤 步骤a）和b），以及d）通过流式细胞术检测用核染色剂处理的测试样品中的微生物的步骤。

公开(公告)号：US20090305240A1

公开(公告)日：2009-12-10

申请号：US11994823

申请日：2006-02-17

申请人： Shinichi Yoshida ,  Takashi Soejima

发明人： Shinichi Yoshida ,  Takashi Soejima

IPC分类号： C12Q1/68

CPC分类号： C12Q1/6806 ,  C12Q1/686 ,  C12Q1/6888 ,  C12Q1/689 ,  C12Q2600/158 ,  C12Q2521/519

摘要： Live cells of a microorganism in a test sample are detected by the following steps: a) the step of treating the test sample with a topoisomerase poison and/or a DNA gyrase poison, b) the step of extracting DNA from the test sample, and amplifying a target region of the extracted DNA by PCR, and c) the step of analyzing an amplification product.

摘要翻译： 通过以下步骤检测测试样品中微生物的活细胞：a）用拓扑异构酶毒物和/或DNA促旋转染酶毒物处理测试样品的步骤，b）从测试样品中提取DNA的步骤，以及 通过PCR扩增提取的DNA的靶区域，和c）分析扩增产物的步骤。

公开(公告)号：US08221975B2

公开(公告)日：2012-07-17

申请号：US12445506

申请日：2008-08-01

申请人： Shinichi Yoshida ,  Takashi Soejima

发明人： Shinichi Yoshida ,  Takashi Soejima

IPC分类号： C12Q1/68

CPC分类号： C12Q1/68 ,  C12Q1/689 ,  C12Q2523/101

摘要： A live cell of microorganism in a test sample is detected by the following steps of: a) adding a cross-linker capable of cross-linking a DNA by irradiation with light having a wavelength of 350 nm to 700 nm to the test sample; b) irradiating the test sample to which the cross-linker is added with light having a wavelength of 350 nm to 700 nm; c) removing the cross-linker contained in the test sample irradiated with light; d) adding a medium to the test sample from which the cross-linker is removed and incubating the test sample; e) adding again the cross-linker capable of cross-linking a DNA by irradiation with light having a wavelength of 350 nm to 700 nm to the incubated test sample; f) irradiating the test sample to which the cross-linker is added with light having a wavelength of 350 nm to 700 nm; g) extracting a DNA from the test sample and amplifying a target region of the extracted DNA by a nucleic acid amplification method; and h) analyzing the amplified product.

摘要翻译： 通过以下步骤检测测试样品中微生物的活细胞：a）通过用波长为350nm至700nm的光照射能够将DNA交联的交联剂加入到测试样品中; b）用波长为350nm至700nm的光照射加入交联剂的测试样品; c）除去用光照射的测试样品中所含的交联剂; d）向测试样品中加入培养基，从中除去交联剂并培养测试样品; e）再次加入能够通过用波长为350nm至700nm的光照射到孵育的测试样品的DNA来交联DNA的交联剂; f）用波长为350nm至700nm的光照射加入交联剂的测试样品; g）从测试样品中提取DNA并通过核酸扩增方法扩增提取的DNA的靶区域; 和h）分析扩增产物。

公开(公告)号：US08026079B2

公开(公告)日：2011-09-27

申请号：US11996422

申请日：2006-02-17

申请人： Shinichi Yoshida ,  Takashi Soejima

发明人： Shinichi Yoshida ,  Takashi Soejima

IPC分类号： C12Q1/02 ,  C12Q1/18

CPC分类号： C12Q1/04 ,  C12Q1/34 ,  C12Q1/533 ,  C12Q2304/10

摘要： According to the following steps, live cells, injured cells, VNC cells and dead cells of a microorganism in a test sample are detected by flow cytometry:a) the step of treating the test sample with an enzyme having an activity of decomposing cells other than those of the microorganism, colloidal particles of proteins or lipids existing in the test sample,b) the step of treating the test sample with a topoisomerase poison and/or a DNA gyrase poison.c) the step of treating the test sample treated in the steps a) and b) with a nuclear stain agent, andd) the step of detecting the microorganism in the test sample treated with the nuclear stain agent by flow cytometry.

摘要翻译： 根据以下步骤，通过流式细胞术检测试验样品中微生物的活细胞，受损细胞，VNC细胞和死细胞：a）用具有分解细胞以外的细胞活性的酶处理测试样品的步骤 微生物的那些，存在于测试样品中的蛋白质或脂质的胶体颗粒，b）用拓扑异构酶毒物和/或DNA回旋酶毒物处理测试样品的步骤，c）处理在测试样品中处理的测试样品的步骤 步骤a）和b），以及d）通过流式细胞术检测用核染色剂处理的测试样品中的微生物的步骤。

公开(公告)号：US20110020821A1

公开(公告)日：2011-01-27

申请号：US12832782

申请日：2010-07-08

申请人： Shinichi Yoshida ,  Takashi Soejima

发明人： Shinichi Yoshida ,  Takashi Soejima

IPC分类号： C12Q1/68 ,  C12Q1/02

CPC分类号： C12Q1/04 ,  C12Q1/34 ,  C12Q1/533 ,  C12Q2304/10

摘要： According to the following steps, live cells, injured cells, VNC cells and dead cells of a microorganism in a test sample are detected by flow cytometry: a) the step of treating the test sample with an enzyme having an activity of decomposing cells other than those of the microorganism, colloidal particles of proteins or lipids existing in the test sample, b) the step of treating the test sample with a topoisomerase poison and/or a DNA gyrase poison. c) the step of treating the test sample treated in the steps a) and b) with a nuclear stain agent, and d) the step of detecting the microorganism in the test sample treated with the nuclear stain agent by flow cytometry.

摘要翻译： 根据以下步骤，通过流式细胞术检测试验样品中微生物的活细胞，受损细胞，VNC细胞和死细胞：a）用具有分解细胞以外的细胞活性的酶处理测试样品的步骤 微生物的那些，存在于测试样品中的蛋白质或脂质的胶体颗粒，b）用拓扑异构酶毒物和/或DNA回旋酶毒物处理测试样品的步骤，c）处理在测试样品中处理的测试样品的步骤 步骤a）和b），以及d）通过流式细胞术检测用核染色剂处理的测试样品中的微生物的步骤。

公开(公告)号：US20110020820A1

公开(公告)日：2011-01-27

申请号：US12832740

申请日：2010-07-08

申请人： Shinichi Yoshida ,  Takashi Soejima

发明人： Shinichi Yoshida ,  Takashi Soejima

IPC分类号： C12Q1/68

CPC分类号： C12Q1/04 ,  C12Q1/34 ,  C12Q1/533 ,  C12Q2304/10

摘要： A kit is disclosed for preparing a measurement sample for detecting live cells, injured cells, VNC cells and dead microorganism cells in a test sample by the following steps: a) the step of treating the test sample with an enzyme having an activity of decomposing cells other than those of the microorganism, colloidal particles of proteins or lipids existing in the test sample, b) the step of treating the test sample with a topoisomerase poison and/or a DNA gyrase poison. c) the step of treating the test sample treated in the steps a) and b) with a nuclear stain agent, and d) the step of detecting the microorganism in the test sample treated with the nuclear stain agent by flow cytometry.

摘要翻译： 公开了用于通过以下步骤制备用于检测测试样品中的活细胞，受损细胞，VNC细胞和死亡微生物细胞的测量样品的试剂盒：a）用具有分解细胞活性的酶处理测试样品的步骤 除了微生物之外，存在于测试样品中的蛋白质或脂质的胶体颗粒，b）用拓扑异构酶毒物和/或DNA回旋酶毒物处理测试样品的步骤，c）处理测试样品的步骤 在步骤a）和b）中使用核染色剂，以及d）通过流式细胞术检测用核染色剂处理的测试样品中的微生物的步骤。

公开(公告)号：US20100170777A1

公开(公告)日：2010-07-08

申请号：US12092677

申请日：2007-07-17

申请人： Shinichi Yoshida ,  Takashi Soejima

发明人： Shinichi Yoshida ,  Takashi Soejima

IPC分类号： C07D221/12 ,  A01N43/42

CPC分类号： A61K31/473 ,  A01N43/42 ,  A61K41/0019

摘要： The present invention provides an agent for degrading a nucleic acid, which includes ethidium monoazide as an active ingredient, and is useful as an antibacterial agent such as a bactericide or a disinfectant.

摘要翻译： 本发明提供了一种降解核酸的试剂，其包括作为有效成分的单脒化合物，可用作抗菌剂如杀菌剂或消毒剂。