Strachan N J, Ogden I D
Department of Plant and Soil Science, University of Aberdeen, Cruickshank Building, Aberdeen, UK.
FEMS Microbiol Lett. 2000 May 1;186(1):79-84. doi: 10.1111/j.1574-6968.2000.tb09085.x.
A microsphere coagulation enzyme-linked immunosorbent assay (MC-ELISA) using Russell's viper venom factor X activator (RVV-XA) is described for the detection of Escherichia coli O157:H7. This microtitre plate assay comprises a standard sandwich immunoassay incorporating RVV-XA as the enzyme label. Coagulation substrate together with polystyrene microspheres are added to the wells of the microtitre plate. RVV-XA initiates the coagulation cascade causing formation of an artificial clot of polystyrene microspheres bound together with fibrin. As few as 10(2) E. coli O157 in a well (10(3) per ml) can be detected within 3 h. The assay is two orders of magnitude more sensitive than a standard ELISA and is a generic technique with the potential for widespread use in sandwich immunoassays.
描述了一种使用罗素蝰蛇毒因子X激活剂（RVV-XA）的微球凝固酶联免疫吸附测定法（MC-ELISA）用于检测大肠杆菌O157:H7。这种微量滴定板测定法包括一种标准的夹心免疫测定法，其中RVV-XA作为酶标记物。将凝血底物与聚苯乙烯微球一起加入到微量滴定板的孔中。RVV-XA启动凝血级联反应，导致形成由与纤维蛋白结合在一起的聚苯乙烯微球组成的人工凝块。每孔中低至10²个大肠杆菌O157（每毫升10³个）在3小时内即可被检测到。该测定法比标准ELISA灵敏两个数量级，并且是一种通用技术，有潜力在夹心免疫测定中广泛应用。
