公开(公告)号：US4495294A

公开(公告)日：1985-01-22

申请号：US441810

申请日：1982-11-15

申请人： Chizuko Nakahara ,  Shigenori Tanaka ,  Hiroshi Tamura ,  Akiyoshi Matsumoto

发明人： Chizuko Nakahara ,  Shigenori Tanaka ,  Hiroshi Tamura ,  Akiyoshi Matsumoto

IPC分类号： G01N33/52 ,  G01N33/50 ,  G01N33/579 ,  C12Q1/00 ,  C12Q1/36 ,  C12Q1/38

CPC分类号： G01N33/579 ,  Y10S435/81 ,  Y10T436/25125

摘要： Bacterial endotoxin in an assay sample is determined by an amoebocyte lysate component of horseshoe crab in the presence or absence of a substrate for endotoxin determination by using an assay sample in the liquid state obtained by treating said assay sample with an endotoxin-free acid having a pKa.sup.25.degree. C. value of not more than 3 at a pH of not more than 3; and a kit therefor, a receptacle containing (a) an endotoxin-free acid having a pKa.sup.25.degree. C. value of not more than 3 e.g. perchloric acid, trichloroacetic acid, nitric acid, for treating the assay sample and a receptacle containing (b) an amoebocyte lysate component or horseshoe crab. The acid treatment of the assay sample results in improved accuracy, reliability and reproducibility of the bacterial endotoxin determination.

摘要翻译： 测定样品中的细菌内毒素通过在存在或不存在用于内毒素测定的底物的存在或不存在下测定的测定样品中的细菌内毒素，其通过使用通过用不含内毒素的酸处理所述测定样品获得的液体状态的测定样品，所述测定样品具有 pKa25℃，pH值不大于3时不大于3; 和一种试剂盒，其容器含有（a）pKa25℃值不大于3的无内毒素的酸，例如， 高氯酸，三氯乙酸，硝酸，用于处理测定样品和含有（b）变形细胞裂解物组分或马蹄蟹的容器。 测定样品的酸处理提高了细菌内毒素测定的准确性，可靠性和再现性。

公开(公告)号：US5876955A

公开(公告)日：1999-03-02

申请号：US774236

申请日：1996-12-27

申请人： Hiroshi Tamura ,  Shigenori Tanaka ,  Jun Aketagawa ,  Toshio Oda

发明人： Hiroshi Tamura ,  Shigenori Tanaka ,  Jun Aketagawa ,  Toshio Oda

IPC分类号： G01N33/579 ,  C12Q1/34 ,  C12N9/99 ,  C12Q1/37

CPC分类号： G01N33/579

摘要： An endotoxin stabilizing agent is provided, which is useful in order to maintain the endotoxin activity of a specimen or a reference standard in a stable state for a prolonged period of time and to prepare an endotoxin reference standard having a reduced intervial variation, being stable for a long time in the form of, in particular, a solution and withstanding repeated use. An endotoxin composition comprising the above-mentioned endotoxin stabilizing agent and endotoxin and a method for assaying endotoxin by using the same are also provided. The endotoxin stabilizing agent comprises: (1) at least one substance selected from the group consisting of aminoalcohols, polyhydric alcohols, nonionic surfactants, polysucroses and chelating agents; (2) at least aminoalcohol and polyhydric alcohol; (3) with the polyhydric alcohol being preferably glycerol or its derivative; (4) polyethylene glycol as an effective ingredient in addition to the components (1) to (3); or (5) at least polysucrose, and an alkaline earth metal salt and/or polyethylene glycol as an effective ingredient. The endotoxin composition comprises the above-identified endotoxin stabilizing agent. The method for assaying endotoxin comprises adding the above-described endotoxin stabilizing agent to a specimen to thereby stabilize the activity of endotoxin in the specimen.

摘要翻译： 提供了一种内毒素稳定剂，其可用于将试样或参考标准品的内毒素活性长时间维持在稳定状态，并制备具有减小的间隙变化的内毒素参照标准物，其对于 很长一段时间的形式，特别是一个解决方案，并承受反复使用。 还提供了包含上述内毒素稳定剂和内毒素的内毒素组合物和使用该内毒素测定内毒素的方法。 所述内毒素稳定剂包括：（1）选自氨基醇，多元醇，非离子表面活性剂，多糖和螯合剂中的至少一种物质; （2）至少氨基醇和多元醇; （3）多元醇优选为甘油或其衍生物; （4）除了组分（1）至（3）之外的聚乙二醇作为有效成分; 或（5）至少聚蔗糖，以及碱土金属盐和/或聚乙二醇作为有效成分。 内毒素组合物包含上述内毒素稳定剂。 用于测定内毒素的方法包括向样品中加入上述内毒素稳定剂，从而稳定样品中内毒素的活性。

公开(公告)号：US5648230A

公开(公告)日：1997-07-15

申请号：US474722

申请日：1995-06-07

申请人： Hiroshi Tamura ,  Shigenori Tanaka ,  Jun Aketagawa ,  Toshio Oda

发明人： Hiroshi Tamura ,  Shigenori Tanaka ,  Jun Aketagawa ,  Toshio Oda

IPC分类号： G01N33/579 ,  C12Q1/34

CPC分类号： G01N33/579

摘要： An endotoxin stabilizing agent is provided, which is useful in order to maintain the endotoxin activity of a specimen or a reference standard in a stable state for a prolonged period of time and to prepare an endotoxin reference standard having a reduced intervial variation, being stable for a long time in the form of, in particular, a solution and withstanding repeated use. An endotoxin composition comprising the above-mentioned endotoxin stabilizing agent and endotoxin and a method for assaying endotoxin by using the same are also provided. The endotoxin stabilizing agent comprises: (1) at least one substance selected from the group consisting of aminoalcohols, polyhydric alcohols, nonionic surfactants, polysucroses and chelating agents; (2) at least aminoalcohol and polyhydric alcohol; (3) with the polyhydric alcohol being preferably glycerol or its derivative; (4) polyethylene glycol as an effective ingredient in addition to the components (1) to (3); or (5) at least polysucrose, and an alkaline earth metal salt and/or polyethylene glycol as an effective ingredient. The endotoxin composition comprises the above-identified endotoxin stabilizing agent. The method for assaying endotoxin comprises adding the above-described endotoxin stabilizing agent to a specimen to thereby stabilize the activity of endotoxin in the specimen.

摘要翻译： 提供了一种内毒素稳定剂，其可用于将试样或参考标准品的内毒素活性长时间维持在稳定状态，并制备具有减小的间隙变化的内毒素参照标准物，其对于 很长一段时间的形式，特别是一个解决方案，并承受反复使用。 还提供了包含上述内毒素稳定剂和内毒素的内毒素组合物和使用该内毒素测定内毒素的方法。 所述内毒素稳定剂包括：（1）选自氨基醇，多元醇，非离子表面活性剂，多糖和螯合剂中的至少一种物质; （2）至少氨基醇和多元醇; （3）多元醇优选为甘油或其衍生物; （4）除了组分（1）至（3）之外的聚乙二醇作为有效成分; 或（5）至少聚蔗糖，以及碱土金属盐和/或聚乙二醇作为有效成分。 内毒素组合物包含上述内毒素稳定剂。 用于测定内毒素的方法包括向样品中加入上述内毒素稳定剂，从而稳定样品中内毒素的活性。

公开(公告)号：US5389547A

公开(公告)日：1995-02-14

申请号：US58306

申请日：1993-05-10

申请人： Shigenori Tanaka ,  Hiroshi Tamura

发明人： Shigenori Tanaka ,  Hiroshi Tamura

IPC分类号： C12Q1/04 ,  C12Q1/37 ,  G01N33/531 ,  G01N33/579 ,  G01N33/00

CPC分类号： G01N33/579 ,  Y10T436/10 ,  Y10T436/14 ,  Y10T436/143333 ,  Y10T436/25 ,  Y10T436/25125

摘要： A reagent denatures or eliminates factors interfering with biochemical reactions by simple treatment without requiring separation of any denatured product precipitate. The reagent makes it possible to assay, in particular, .beta.-glucan and endotoxin in blood-derived samples rapidly and efficiently with high sensitivity. The reagent includes a hexadimethrine compound and an alkali metal hydroxide or an alkali metal hydroxide as a main component. A method for assaying a substance specifically reacting with a Limulus reagent utilizing the reagent, an assay kit including at least the reagent and a Limulus reagent, and a method of diagnosing infectious diseases based on the results obtained by the assay method are also provided.

摘要翻译： 试剂通过简单的处理变性或消除干扰生化反应的因素，而不需要分离任何变性产物沉淀物。 该试剂使得可以以高灵敏度快速有效地测定血液来源的样品中的β-葡聚糖和内毒素。 该试剂含有己二胺化合物和碱金属氢氧化物或碱金属氢氧化物作为主要成分。 还提供了一种使用试剂测定与鲎试剂特异性反应的物质的方法，至少包含试剂和鲎试剂的测定试剂，以及基于通过测定方法获得的结果诊断感染性疾病的方法。

公开(公告)号：US5378610A

公开(公告)日：1995-01-03

申请号：US859398

申请日：1992-05-28

申请人： Shigenori Tanaka ,  Hiroshi Tamura

发明人： Shigenori Tanaka ,  Hiroshi Tamura

IPC分类号： G01N33/579 ,  C12Q1/34 ,  C12Q1/00 ,  C12Q1/02

CPC分类号： G01N33/579

摘要： The present invention has been made in order to solve the problems in conventional methods of assaying an endotoxin in a specimen such as plasma or serum by the limulus test, which requires a complicated pretreatment procedure such as centrifugation for removing denatured precipitates formed with an acid treatment. According to the assay method of the present invention, an endotoxin adsorbed by proteins, lipids and platelets can be efficiently liberated simply by adding a mixed aqueous solution having a specific composition according to the present invention without any separation procedure and thus a sample solution of good qualities can be prepared. After adding the mixed aqueous solution containing a specific surfactant, a compound having an imidazolyl group or an amino group and an alkaline earth metal salt and an alkaline metal hydroxide to the specimen, followed by addition of the limulus amoebocyte lysate components, the endotoxin can be easily, quickly and precisely assayed. In particular, gram-negative bacterial septicemia, which can be hardly diagnosed, can be quickly assayed thereby.

摘要翻译： PCT No.PCT / JP91 / 01309 Sec。 371日期：1992年5月28日 102（e）日期1992年5月28日PCT 1991年9月27日PCT公布。 出版物WO92 / 06382 日期为1992年4月16日。本发明是为了解决通过鲎试验来测定血浆或血清等标本中的内毒素的常规方法中的问题，需要复杂的预处理步骤，例如离心分离 用酸处理形成变性沉淀。 根据本发明的测定方法，可以简单地通过添加根据本发明的具有特定组成的混合水溶液而无需任何分离程序，有效地释放被蛋白质，脂质和血小板吸附的内毒素，因此，可以有良好的样品溶液 品质可以做好准备。 将含有特定表面活性剂的混合水溶液，具有咪唑基或氨基的化合物和碱土金属盐和碱金属氢氧化物加入到样品中，然后加入鲎变形细胞裂解物组分，内毒素可以是 容易，快速和准确地测定。 特别地，可以快速测定可以很难诊断的革兰氏阴性细菌性败血症。

公开(公告)号：US06306577B1

公开(公告)日：2001-10-23

申请号：US09290091

申请日：1999-04-12

申请人： Hiroshi Tamura ,  Shigenori Tanaka

发明人： Hiroshi Tamura ,  Shigenori Tanaka

IPC分类号： C12Q100

CPC分类号： C12Q1/00 ,  G01N33/579 ,  G01N2333/90219 ,  G01N2333/90245 ,  Y02A90/26 ,  Y10S435/963

摘要： A method for measuring an enzyme reaction to determine an amount of a substance involved in the enzyme reaction, which comprises measuring a time course of a parameter of the enzyme reaction, measuring a time required for the parameter of the enzyme reaction to change from a first threshold value to a second threshold value, and correlating the measured time to an amount of the substance involved in the enzyme reaction.

摘要翻译： 一种测定酶反应以测定酶反应参与物质的量的方法，该方法包括测定酶反应参数的时间过程，测定酶反应参数所需的时间从第一 阈值至第二阈值，并且将测量的时间与酶反应中涉及的物质的量相关联。

公开(公告)号：US06284885B1

公开(公告)日：2001-09-04

申请号：US09485714

申请日：2000-03-01

申请人： Hiroshi Tamura ,  Maki Aizawa ,  Shigenori Tanaka

发明人： Hiroshi Tamura ,  Maki Aizawa ,  Shigenori Tanaka

IPC分类号： C07H100

CPC分类号： C08B37/0024

摘要： The present invention relates to a process for preparing (1→3)-&bgr;-D-glucan, comprising subjecting a fungal cell to oxidation degradation under alkaline conditions to release (1→3)-&bgr;-D-glucan from a cell wall of the cell. (1→3)-&bgr;-D-Glucan can be prepared easily and efficiently from a cell of a microorganism belonging to the genus Candida. An accurate measurement process having a high reproducibility of (1→3)-&bgr;-D-glucan can be provided by using the above-described glucan or stabilized glucan into a kit and using it as a standard substance, and a large amount of the (1→3)-&bgr;-D-glucan having various biological activities can be solubilized and purified from a fungus, such as Candida or the like.

摘要翻译： 本发明涉及一种制备（1-> 3）-β-D-葡聚糖的方法，包括在碱性条件下使真菌细胞进行氧化降解以从细胞中释放（1-> 3）-β-D-葡聚糖 细胞壁。 （1-> 3）-β-D-葡聚糖可以从属于念珠菌属的微生物的细胞中容易且有效地制备。 通过使用上述葡聚糖或稳定的葡聚糖作为试剂盒并使用其作为标准物质，可以提供具有（1-> 3）-β-D-葡聚糖的高再现性的精确测量方法，并将大量 具有各种生物活性的（1-> 3）-β-D-葡聚糖可以从真菌如假丝酵母等中溶解和纯化。

公开(公告)号：US5702882A

公开(公告)日：1997-12-30

申请号：US313899

申请日：1994-09-28

申请人： Hiroshi Tamura ,  Toshio Oda ,  Shigenori Tanaka

发明人： Hiroshi Tamura ,  Toshio Oda ,  Shigenori Tanaka

IPC分类号： G01N33/579 ,  C12Q1/00 ,  C12Q1/34 ,  C12Q1/56 ,  G01N33/53

CPC分类号： G01N33/579 ,  Y10S435/975

摘要： Endotoxin (Et) can be specifically assayed by exclusively utilizing the factor C system reaction without being affected by factor G contained in a limulus amebocyte lysate reagent. The present invention provides: (1) a reagent for ET-specific assay which comprises a limulus amebocyte lysate reagent and an alkylglucoside; (2) a method of specifically assaying Et in a specimen using a limulus amebocyte lysate reagent, wherein an alkylglucoside is added to the limulus amebocyte lysate reagent and/or the specimen; and (3) a factor G activation inhibitor composition which comprises an alkylglucoside as an active ingredient capable of inhibiting the activation of factor G in limulus amebocyte by (1.fwdarw.3)-.beta.-D-glucan.

摘要翻译： 内毒素（Et）可以通过专门利用因子C系统反应而不受鲎变形细胞裂解物试剂中所含的因子G的影响而被特异性地测定。 本发明提供：（1）用于ET特异性测定的试剂，其包含鲎变形细胞裂解物试剂和烷基葡糖苷; （2）使用鲎变形细胞裂解物试剂特异性测定试样中的Et的方法，其中将烷基葡糖苷加入到鲎变形细胞裂解物试剂和/或试样中; （3）一种因子G激活抑制剂组合物，其包含烷基葡糖苷作为活性成分，其能够通过（1-> 3）-β-D-葡聚糖抑制鲎变形细胞中因子G的活化。

公开(公告)号：US6156519A

公开(公告)日：2000-12-05

申请号：US793679

申请日：1997-06-02

申请人： Hiroshi Tamura ,  Shigenori Tanaka

发明人： Hiroshi Tamura ,  Shigenori Tanaka

IPC分类号： C07K14/435 ,  C07K16/18 ,  G01N33/579 ,  G01N33/53

CPC分类号： G01N33/579 ,  C07K14/43509 ,  C07K16/18 ,  G01N2400/24 ,  Y10S435/962 ,  Y10S530/857

摘要： A (1.fwdarw.3)-.beta.-D-glucan binding protein obtainable by affinity chromatography or gel filtration of an extract of horseshoe crab amoebocytes and having a molecular weight of about 580 kDa, as determined by gel filtration under non-reducing conditions, and about 170 kDa, as determined by SDS-PAGE under reducing conditions, is provided.An antibody which selectively recognizes the protein is also provided. Methods for detecting or removing (1.fwdarw.3)-.beta.-D-glucan in or from a sample using the protein and antibody are disclosed.

摘要翻译： PCT No.PCT / JP95 / 01735 Sec。 371日期：1997年6月2日 102（e）日期1997年6月2日PCT提交1995年8月31日PCT公布。 出版物WO96 / 06858 日期：1996年3月7日A（1-> 3）-β-D-葡聚糖结合蛋白，可通过亲和层析或凝胶过滤马蹄蟹变形细胞提取物获得，分子量约580kDa，通过凝胶过滤测定 提供非还原条件，约170kDa，通过SDS-PAGE在还原条件下测定。 还提供了选择性识别蛋白质的抗体。 公开了使用蛋白质和抗体检测或从样品中除去（1-> 3）-β-D-葡聚糖的方法。

公开(公告)号：US5681710A

公开(公告)日：1997-10-28

申请号：US391097

申请日：1995-02-21

申请人： Shigenori Tanaka ,  Hiroshi Tamura ,  Makoto Ohki

发明人： Shigenori Tanaka ,  Hiroshi Tamura ,  Makoto Ohki

IPC分类号： G01N33/579 ,  C12Q1/56 ,  C12Q1/00 ,  C12Q1/34 ,  C12Q1/44

CPC分类号： G01N33/579 ,  G01N2400/22 ,  G01N2400/24

摘要： The present invention provides a reagent for determining (1.fwdarw.3)-.beta.-D-glucan comprising a lysate substantially free from any endotoxin-sensitive factor, which is obtained by contacting limulus amebocyte lysate optionally containing dextran with a polyamide or cellulose insoluble carrier, which makes it possible to rapidly and easily determine at a high accuracy (1.fwdarw.3)-.beta.-D-glucan of mycotic origin contained in the body fluid such as blood or urine.

摘要翻译： 本发明提供了一种用于测定（1→3）-β-D-葡聚糖的试剂，其包含基本上不含任何内毒素敏感因子的裂解物，所述裂解物通过任选地含有葡聚糖的鲎变形细胞裂解物与聚酰胺或纤维素不溶性载体 ，这使得可以以高精度（1→3）-β-D-葡聚糖快速和容易地测定包含在体液如血液或尿液中的真菌来源。