公开(公告)号：US3990947A

公开(公告)日：1976-11-09

申请号：US556504

申请日：1975-03-07

申请人： James R. Butler ,  Walter E. Jacobson ,  Donald Paul Kronish ,  James E. Turner ,  Lee S. Zuriff

发明人： James R. Butler ,  Walter E. Jacobson ,  Donald Paul Kronish ,  James E. Turner ,  Lee S. Zuriff

IPC分类号： G01N33/68 ,  C12Q1/56 ,  C12K1/04 ,  G01N33/00

CPC分类号： C12Q1/56 ,  G01N2333/31 ,  G01N2333/75 ,  Y10S435/883

摘要： An improved composition and method for detecting fibrinogen, fibrinogen split products and/or fibrin split products in blood comprises utilizing killed, dyed Staphylococcus aureus cells which can be prepared by either of two methods. In the first method, Staphylococcus aureus organisms are incubated in a nutrient medium containing a polyalkylene glycol having a molecular weight of from 1000 to 5000 to which is added triphenyltetrazolium chloride; the growing organisms reduce the triphenyltetrazolium chloride to triphenylformazan which imparts coloration to the cells; the dyed Staphylococcus aureus cells are killed and substantially all untrapped dye is removed. In an alternate method, a suspension of Fast Black Salt K is added to a suspension of killed Staphylococcus aureus cells, and the dyed cells which result are washed to remove substantially all unfixed dye. The killed, dyed Staphylococcus aureus cells prepared by either method are suspended in an imidazole buffer to maintain a pH of 7.4. The suspension may be lyophilized if desired. The determination of fibrinogen split products and/or fibrin split products in blood serum is performed by serially diluting the blood serum test sample and adding a specified volume of reconstituted killed, dyed Staphylococcus aureus cells to an equivalent volume of each serial dilution of serum test sample, and observing the serial dilutions for the presence of visible clumping as a positive test result. Coloration of the cells greatly improves visualization of the end point of the test. The concentration of fibrinogen split products and/or fibrin split products in the blood serum test sample can be calculated if a comparison test is run with a serum containing a known amount of fibrinogen, fibrinogen split products and/or fibrin split products.

摘要翻译： 用于检测血液中纤维蛋白原，纤维蛋白原分裂产物和/或血纤蛋白分离产物的改进的组合物和方法包括使用可以通过两种方法中的任一种制备的杀死的，染色的金黄色葡萄球菌细胞。 在第一种方法中，将金黄色葡萄球菌生物体在含有分子量为1000-5000的聚亚烷基二醇的营养培养基中温育，加入三苯基四氮唑鎓氯化物; 生长的生物体将三苯基四唑氯化物还原成三苯甲脒，使细胞着色; 染色的金黄色葡萄球菌细胞被杀死并且基本上除去所有未捕获的染料。 在另一种方法中，将快速黑盐K的悬浮液加入到金黄色葡萄球菌细胞的悬浮液中，洗涤出染色的细胞以除去基本上所有未固定的染料。 将通过任一方法制备的经灭活的染色的金黄色葡萄球菌细胞悬浮在咪唑缓冲液中以保持pH为7.4。 如果需要，悬浮液可以冻干。 血清中纤维蛋白原分裂产物和/或纤维蛋白裂解产物的测定通过连续稀释血清测试样品并将特定体积的重构杀死的染色的金黄色葡萄球菌细胞加入等体积的每个连续稀释的血清测试样品 并观察连续稀释度作为阳性测试结果的可见聚集物的存在。 细胞的着色大大提高了测试终点的可视化。 如果使用含有已知量的纤维蛋白原，纤维蛋白原分裂产物和/或纤维蛋白裂解产物的血清进行比较测试，则可以计算血清测试样品中纤维蛋白原裂解产物和/或纤维蛋白裂解产物的浓度。