Phenotypic and genotypic typing of food and clinical isolates of Enterobacter sakazakii

Enterobacter sakazakii, designated a unique species in 1980, has been impli cated as the causative organism in a rare but severe form of neonatal menin gitis. Dried infant formula milk has been identified as a potential source of the organism, E. sakazakii isolates from dried infant formula available in Canada and clinical isolates obtained from Canadian hospital culture col lections were characterised by phenotypic (biotype and antibiograms) and ge notypic (ribotyping, random amplification of polymorphic DNA and pulsed-fie ld gel electrophoresis) methods. Three biotypes and four antibiogram patter ns were observed in the 18 isolates examined. Ribotyping with the Dupont Ri boprinter(TM) microbial identification system divided the 18 isolates into 10 ribotypes. Three isolates from the same hospital bah indistinguishable r ibotyping patterns although each was isolated in a different year, as did t hree food isolates from one company Pulsed-field gel electrophoresis (PFGE) and random amplification of polymorphic DNA (RAPD) profiles indicated mino r differences between the isolates that were indistinguishable by ribotypin g, PFGE (with the restriction endonucleases Xba1 and Spe1) and RAPD gave di screte patterns that enabled easy comparison of E, sakazakii isolates, with a high degree of discrimination. The discriminatory index showed RAPD and PFGE were shown to be the most discriminatory typing schemes for E, sakazak ii, followed by ribotyping, biotyping and antibiograms.