公开(公告)号：US5856117A

公开(公告)日：1999-01-05

申请号：US879962

申请日：1997-06-20

申请人： Harumi Uenoyama ,  Kyouichi Ohshiro ,  Atsuko Nanbu ,  Satoshi Fukunaga

发明人： Harumi Uenoyama ,  Kyouichi Ohshiro ,  Atsuko Nanbu ,  Satoshi Fukunaga

IPC分类号： C12Q1/37 ,  C12Q1/00 ,  G01N33/53

CPC分类号： C12Q1/37 ,  C12Q2337/12 ,  G01N2333/81 ,  G01N2333/811 ,  G01N2333/976 ,  Y10S435/963 ,  Y10S435/975

摘要： This invention provides a method for measuring the concentration of urinary trypsin inhibitors which is excellent in precision and reproducibility, whose operation is simple, and in which a possibility of damaging a plastic cell is eliminated. The method for measuring the concentration of urinary trypsin inhibitors comprises mixing an urine sample, a protease solution containing trypsin, and a buffer solution, adding a substrate solution to the mixture to cause the enzyme reaction, and measuring the activity of the enzyme, wherein the buffer solution is prepared so that it contains at least 0.15 .mu.mol calcium per 1 .mu.g of the trypsin but no more than 100 .mu.mol calcium per 1 ml of the urine sample in the reaction mixture, and wherein the substrate solution is prepared by dissolving the substrate in an organic solvent and diluting the mixture solution with aqueous medium, wherein at least one of an amphoteric surfactant and a nonionic surfactant is added to at least one of the organic solvent and the aqueous medium.

摘要翻译： 本发明提供了一种测定尿胰蛋白酶抑制剂浓度的方法，其精度和重复性优异，操作简单，消除了塑料细胞损伤的可能性。 用于测定尿胰蛋白酶抑制剂浓度的方法包括混合尿液样品，含有胰蛋白酶的蛋白酶溶液和缓冲溶液，向混合物中加入底物溶液以引起酶反应，并测量酶的活性，其中 制备缓冲液，使其每1ml胰蛋白酶含有至少0.15μmol钙，而在反应混合物中每1ml尿液含有不超过100μmol钙，其中底物溶液通过溶解 将所述底物置于有机溶剂中并用水性介质稀释所述混合溶液，其中至少一种两性表面活性剂和非离子表面活性剂加入至少一种有机溶剂和水性介质中。

公开(公告)号：US6130055A

公开(公告)日：2000-10-10

申请号：US135915

申请日：1998-08-18

申请人： Atsuko Nanbu ,  Satoshi Fukunaga

发明人： Atsuko Nanbu ,  Satoshi Fukunaga

IPC分类号： G01N33/15 ,  C12Q1/37 ,  C12Q1/34

CPC分类号： C12Q1/37 ,  C12Q2337/12 ,  G01N2333/811 ,  G01N2333/8114 ,  G01N2333/976 ,  Y10S435/962

摘要： A method for measuring the concentration or the activity of UTI quickly and easily at high sensitivity. A urine sample, a buffer solution, a trypsin solution and a substrate solution are mixed and the trypsin activity is then measured. Thus, the UTI concentration in the urine sample is determined. In this case, a substrate solution having only L-BAPNA is used as the substrate, and the surfactant is mixed in at least one selected from the buffer solution and the enzyme solution. The mixing ratio of the surfactant is about 1 wt. % in the entire enzyme reaction solution. Examples of the surfactant include polyoxyethylene (40) octylphenylether, polyoxyethylene (10) octylphenylether, 3-[(3-cholamido propyl)dimethylammonio]-propanesulfonic acid, 3-[(3-cholamido propyl)dimethylammonio]-2-hydroxypropanesulfonic acid, and polyoxyethylene sorbitan monolaurate. As shown in FIG. 1, the sensitivity improves when using L-BAPNA.

摘要翻译： 用于以高灵敏度快速，容易地测量UTI的浓度或活性的方法。 将尿样，缓冲溶液，胰蛋白酶溶液和底物溶液混合，然后测定胰蛋白酶活性。 因此，测定尿样中的UTI浓度。 在这种情况下，使用仅具有L-BAPNA的底物溶液作为底物，并且将表面活性剂混合在选自缓冲溶液和酶溶液中的至少一种。 表面活性剂的混合比为约1wt。 ％在整个酶反应溶液中。 表面活性剂的实例包括聚氧乙烯（40）辛基苯基醚，聚氧乙烯（10）辛基苯基醚，3 - [（3-胆酰胺基丙基）二甲基氨基] - 丙磺酸，3 - [（3-胆酰胺基丙基）二甲基氨基] -2-羟基丙磺酸和 聚氧乙烯脱水山梨醇单月桂酸酯。 如图所示。 1，使用L-BAPNA时灵敏度提高。