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    • 4. 发明授权
    • Endotoxin stabilizing agent, endotoxin composition and method for
assaying endotoxin
    • 内毒素稳定剂,内毒素组成和测定内毒素的方法
    • US5876955A
    • 1999-03-02
    • US774236
    • 1996-12-27
    • Hiroshi TamuraShigenori TanakaJun AketagawaToshio Oda
    • Hiroshi TamuraShigenori TanakaJun AketagawaToshio Oda
    • G01N33/579C12Q1/34C12N9/99C12Q1/37
    • G01N33/579
    • 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.
    • 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。
    • 5. 发明授权
    • Endotoxin stabilizing agent, endotoxin composition and method for
assaying endotoxin
    • 内毒素稳定剂,内毒素组成和测定内毒素的方法
    • US5648230A
    • 1997-07-15
    • US474722
    • 1995-06-07
    • Hiroshi TamuraShigenori TanakaJun AketagawaToshio Oda
    • Hiroshi TamuraShigenori TanakaJun AketagawaToshio Oda
    • G01N33/579C12Q1/34
    • G01N33/579
    • 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.
    • 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。
    • 7. 发明授权
    • Reagent for endotoxin-specific assay
    • 内毒素特异性检测试剂
    • US5550030A
    • 1996-08-27
    • US120469
    • 1993-09-14
    • Shigenori TanakaHiroshi TamuraMakoto Ohki
    • Shigenori TanakaHiroshi TamuraMakoto Ohki
    • G03C5/50G01N33/545G01N33/548G01N33/579G03C7/407C12Q1/37C12N11/08C12N11/12C12Q1/34
    • G01N33/579
    • Disclosed are a reagent for endotoxin-specific assay which comprises an insoluble carrier having immobilized thereon at least an endotoxin-sensitive factor derived from a limulus amebocyte; a kit for endotoxin-specific assay containing said reagent and a substrate for activated factor C or a substrate for clotting enzyme; a method for assaying endotoxin comprising applying a sample solution to said reagent to cause endotoxin in the sample to react with factor C in said reagent and determining a change of a substrate; and a process for preparing said reagent which comprises physically or chemically immobilizing at least an endotoxin-sensitive factor derived from a limulus amebocyte on an insoluble carrier. Endotoxin in a sample, even turbid or colored, can be specifically assayed with ease and rapidness without the influence of a (1.fwdarw.3)-.beta.-glucan.
    • 公开了一种用于内毒素特异性测定的试剂,其包含其上固定有至少一种源自鲎变形细胞的内毒素敏感因子的不溶性载体; 含有所述试剂和活化因子C的底物或用于凝固酶的底物的内毒素特异性测定试剂盒; 一种用于测定内毒素的方法,包括将样品溶液施加到所述试剂以使样品中的内毒素与所述试剂中的因子C反应并确定底物的变化; 以及制备所述试剂的方法,其包括在不溶性载体上物理或化学地固定来自鲎变形细胞的至少一种内毒素敏感因子。 样品中的内毒素,甚至浑浊或有色,可以在不受(1-> 3)-β-葡聚糖的影响的情况下容易且快速地进行特异性测定。
    • 9. 发明授权
    • Method for assaying endotoxin in serum or plasma using limulus
amoebocyte lysate
    • 使用鲎变形细胞裂解液测定血清或血浆中的内毒素的方法
    • US5378610A
    • 1995-01-03
    • US859398
    • 1992-05-28
    • Shigenori TanakaHiroshi Tamura
    • Shigenori TanakaHiroshi Tamura
    • G01N33/579C12Q1/34C12Q1/00C12Q1/02
    • G01N33/579
    • 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.
    • 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。