Open AccessA sensitive microsphere coagulation ELISA for Escherichia coli O157:H7 using Russell's viper venom
Author(s) -
Strachan Norval J.C.,
Ogden Iain D.
Publication year - 2000
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.2000.tb09085.x
Subject(s) - viper venoms , chemistry , immunoassay , escherichia coli , coagulation , chromatography , venom , polystyrene , microbiology and biotechnology , snake venom , biology , antibody , biochemistry , immunology , medicine , organic chemistry , psychiatry , gene , polymer
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.