公开(公告)号：US20080014578A1

公开(公告)日：2008-01-17

申请号：US10584393

申请日：2004-12-24

申请人： Naoko Horikoshi ,  Susumu Kawasaki ,  Yukio Okada ,  Kazuko Takeshita ,  Takashi Sameshima ,  Shinichi Kawamoto ,  Kenji Isshiki

发明人： Naoko Horikoshi ,  Susumu Kawasaki ,  Yukio Okada ,  Kazuko Takeshita ,  Takashi Sameshima ,  Shinichi Kawamoto ,  Kenji Isshiki

IPC分类号： C12Q1/68

CPC分类号： C12Q1/686 ,  C12Q1/689 ,  Y02A50/451

摘要： The present invention is to provide a multiple detection method that can detect contaminating microorganisms existing in foods, including pathogenic Escherichia coli O157, Listeria monocytogenes and Salmonella spp., with high sensitivity comparable or even superior to official methods, comprising the steps of amplifying a plural number of target genes with a single PCR reaction tube and analyzing the same. The following steps are performed consecutively: (A) a step of extracting DNA of the target microorganisms to be detected by treating with at least a lytic enzyme such as Achromopepidase and Lyzocyme and/or bacteriocin having lytic activity such as Enterolysine, a surfactant and a protein denaturing agent; and (B) a step of mixing a specific primer to the target microorganisms to be detected to perform multiplex PCR. Further, it is preferable to add a step of culturing with a culture condition where 1 CFU/100 g microorganisms becomes 103 CFU/ml or more after 18 to 48 h of culture, for example that the pH after culture becomes 5.1 or more, before the step of extracting DNA of the target microorganisms to be detected.

摘要翻译： 本发明提供一种可以检测存在于食品中的污染微生物的多重检测方法，包括致病性大肠杆菌O157，单核细胞增生李斯特氏菌和沙门氏菌，具有与官方方法相当或甚至优于其他方法的高灵敏度，包括以下步骤： 使用单个PCR反应管的目标基因数目并进行分析。 连续进行以下步骤：（A）通过用至少溶解酶如具有溶解活性的溶色酶和溶菌酶等溶菌酶和表面活性剂和表面活性剂的溶解活性的溶菌酶和/或细菌素进行处理来提取要检测的目标微生物的DNA的步骤 蛋白变性剂; 和（B）将特异性引物与要检测的目标微生物混合以进行多重PCR的步骤。 此外，优选在培养18〜48小时后，加入培养条件，其中1CFU / 100g微生物变为10 3 CFU / ml以上，例如pH 在提取要检测的目标微生物的DNA的步骤之前，培养变为5.1以上。

公开(公告)号：US06242208B1

公开(公告)日：2001-06-05

申请号：US08235238

申请日：1994-05-02

申请人： Yasushi Shirahase ,  Kenji Isshiki ,  Yoshifumi Watazu

发明人： Yasushi Shirahase ,  Kenji Isshiki ,  Yoshifumi Watazu

IPC分类号： C12Q132

CPC分类号： C12Q1/32

摘要： By a process comprising inhibiting LDH2, LDH3, LDH4 and LDH5 in a sample with a protease in the presence of a protein-denaturating agent and then determining LDH1 remaining uninhibited, LDH1 in the sample is easily determined.

摘要翻译： 通过包括在蛋白质变性剂存在下用蛋白酶抑制LDH2，LDH3，LDH4和LDH5，然后测定LDH1保持不受抑制的方法，容易确定样品中的LDH1。

公开(公告)号：US08298758B2

公开(公告)日：2012-10-30

申请号：US10584393

申请日：2004-12-24

申请人： Naoko Horikoshi ,  Susumu Kawasaki ,  Yukio Okada ,  Kazuko Takeshita ,  Takashi Sameshima ,  Shinichi Kawamoto ,  Kenji Isshiki

发明人： Naoko Horikoshi ,  Susumu Kawasaki ,  Yukio Okada ,  Kazuko Takeshita ,  Takashi Sameshima ,  Shinichi Kawamoto ,  Kenji Isshiki

IPC分类号： C12Q1/68

CPC分类号： C12Q1/686 ,  C12Q1/689 ,  Y02A50/451

摘要： The present invention is to provide a multiple detection method that can detect contaminating microorganisms existing in foods, including pathogenic Escherichia coli O157, Listeria monocytogenes and Salmonella spp., with high sensitivity comparable or even superior to official methods, comprising the steps of amplifying a plural number of target genes with a single PCR reaction tube and analyzing the same. The following steps are performed consecutively: (A) a step of extracting DNA of the target microorganisms to be detected by treating with at least a lytic enzyme such as Achromopepidase and Lysozyme and/or bacteriocin having lytic activity such as Enterolysine, a surfactant and a protein denaturing agent; and (B) a step of mixing a specific primer to the target microorganisms to be detected to perform multiplex PCR. Further, it is preferable to add a step of culturing with a culture condition where 1 CFU/100 g microorganisms becomes 10.sup.3 CFU/ml or more after 18 to 48 h of culture, for example that the pH after culture becomes 5.1 or more, before the step of extracting DNA of the target microorganisms to be detected.

摘要翻译： 本发明提供一种可以检测存在于食品中的污染微生物的多重检测方法，包括致病性大肠杆菌O157，单核细胞增生李斯特氏菌和沙门氏菌，具有与官方方法相当或甚至优于其他方法的高灵敏度，包括以下步骤： 使用单个PCR反应管的目标基因数目并进行分析。 连续进行以下步骤：（A）通过用至少一种裂解酶如溶酶酶和溶菌酶和/或具有溶解活性的细菌素（例如肠溶素，表面活性剂和表面活性剂）进行处理来提取要检测的目标微生物的DNA的步骤 蛋白变性剂; 和（B）将特异性引物与要检测的目标微生物混合以进行多重PCR的步骤。 此外，优选在培养18〜48小时后，添加培养条件，其中1CFU / 100g微生物变成10μFCFU / ml以上，例如培养后的pH变为5.1 或更多，在提取要检测的目标微生物的DNA的步骤之前。

公开(公告)号：US20050214884A1

公开(公告)日：2005-09-29

申请号：US10995383

申请日：2004-11-24

申请人： Yasuhiro Sakai ,  Kenji Isshiki ,  Kouji Kishi

发明人： Yasuhiro Sakai ,  Kenji Isshiki ,  Kouji Kishi

IPC分类号： C12N9/02 ,  C12N9/04 ,  C12N9/96 ,  C12Q1/32 ,  C12Q1/60 ,  G01N33/92

CPC分类号： C12N9/96 ,  C12Q1/60

摘要： The present invention is to provide a novel method for stabilizing cholesterol dehydrogenase. Also, the present invention is to provide a novel cholesterol-containing composition and a cholesterol measuring reagent.

摘要翻译： 本发明提供一种稳定胆固醇脱氢酶的新方法。 另外，本发明提供一种新型含胆固醇的组合物和胆固醇测定试剂。

公开(公告)号：US20050003051A1

公开(公告)日：2005-01-06

申请号：US10500870

申请日：2003-02-06

申请人： Kenji Isshiki ,  Junzo Ogawa

发明人： Kenji Isshiki ,  Junzo Ogawa

IPC分类号： C12Q1/04 ,  C12Q1/22 ,  G01N33/02

CPC分类号： C12Q1/22 ,  C12Q1/04 ,  G01N33/025

摘要： There are provided a method of evaluating a quality of foods and drinks and an indicator therefor in which the quality of the foods and drinks, particularly the lowering of freshness, i.e. an amount of microorganism proliferated in the foods and drinks can be detected simply and accurately to judge the quality. That is, an aerogen consisting of at least one of yeast, mold and bacteria is sealed together with a fermentation substrate in a closed transparent vessel or transparent soft film bag of a synthetic resin and a quality of foods and drinks is judged by an amount of a gas generated through the formation of an acid generated in the vessel (bag), and there is used an indicator for evaluating thereof.

摘要翻译： 提供了一种评价食品和饮料质量的方法及其指标，其中可以简单且准确地检测食品和饮料的质量，特别是降低新鲜度，即食物和饮料中增殖的微生物的量 判断质量。 也就是说，将由酵母，霉菌和细菌中的至少一种构成的造气体与发酵基材一起在封闭的透明容器或合成树脂的透明软膜袋中密封，食品和饮料的质量由 通过形成在容器（袋）中产生的酸而产生的气体，并且使用用于评价的指示剂。