Glucuronidase assay in a rapid MPN determination for recovery of Escherichia coli from selected foods.
Glucuronidase is present in most strains of Escherichia coli but absent in most other enteric microorganisms; therefore, an assay for this enzyme is useful for determining the presence of the organism. The substrate 4-methylumbelliferyl beta-D-glucuronide (MUG) is incorporated into either lauryl tryptose (LT) broth or EC medium; the inoculated tubes are then incubated under specified conditions and examined under longwave UV light for the presence of a fluorogenic glucuronidase end product. When compared with the 10-day most probable number (MPN) procedure of AOAC, the LT-MUG and the EC-MUG tests required 24 and 96 h, respectively, and gave comparable mean log MPN values for samples of crabmeat, sunflower kernels, and walnut pieces. However, false-positive and false-negative reactions were observed with foods tested by both of these rapid methods. Overall, method sensitivity was not compromised by using the LT-MUG rather than the EC-MUG method. Incorporation of 25 micrograms MUG/mL into LT broth resulted in diminished fluorescence of positive reactions, whereas MUG concentrations of 50 and 100 micrograms/mL provided decisive fluorogenic reactions.
D003470:Culture Media; D004926:Escherichia coli / Q000302:isolation & purification*; D005516:Food Microbiology; D005966:Glucuronidase / Q000032:analysis*; D006923:Hymecromone / Q000031:analogs & derivatives; D013050:Spectrometry, Fluorescence
D003470:Culture Media; D006923:Hymecromone; C034888:4-methylumbelliferyl glucuronide; D005966:Glucuronidase