Fluorogenic assay for rapid detection of Escherichia coli in food.
An assay procedure to screen for Escherichia coli in foods by using 4-methylumbelliferyl-beta-D-glucuronide (MUG) incorporated into lauryl tryptose (LST) broth was evaluated. The beta-glucuronidase produced by E. coli cleaves the MUG substrate to yield a fluorescent end product. E. coli-negative samples can be identified by lack of fluorescence in LST-MUG within 24 h. MUG was not inhibitory to coliforms and E. coli. Over 1,400 food and dairy samples were tested to compare the standard three-tube most-probable-number procedure with the MUG-containing or non-MUG-containing LST procedure. LST-MUG testing detected a greater number of E. coli, with a lower false-positive rate (1.4%) and in a shorter time, than did the standard procedure. All false-positive results in the LST-MUG testing were attributable to beta-glucuronidase-producing staphylococci. No false-negative result was encountered. Use of MUG in LST broth obviates the EC broth step, allowing a 2.5-day procedure to a completed E. coli test versus the present 4- to 6-day standard most-probable-number method.
D001431:Bacteriological Techniques; D004926:Escherichia coli / Q000201:enzymology / Q000302:isolation & purification*; D005456:Fluorescent Dyes; D005516:Food Microbiology; D005966:Glucuronidase / Q000032:analysis; D006923:Hymecromone / Q000031:analogs & derivatives
D005456:Fluorescent Dyes; D006923:Hymecromone; C034888:4-methylumbelliferyl glucuronide; D005966:Glucuronidase