Rapid diagnosis of enterohemorrhagic Escherichia coli O157:H7 directly from fecal specimens using immunofluorescence stain.

Abstract

Serotype O157:H7 is most frequently encountered among verotoxin-producing Escherichia coli. Most laboratories use MacConkey-sorbitol agar as a screening medium. Presumptive identification of sorbitol-negative colonies is then accomplished by latex agglutination or biochemical tests with serologic confirmation, which requires 18-36 hours for completion. This study attempted to detect E coli O157:H7 directly from stool specimens by direct immunofluorescence (DIF) antibody staining to provide quicker turnaround (< 2 hours). A total of 336 abnormal fecal samples (bloody, watery, semi-liquid, or mucoid) were examined by this method. Results were compared with those of culture. Direct immunofluorescence antibody staining of the direct fecal smear detected all isolates of E coli O157 that were recovered by culture, including nonmotile strains, strains possessing the H7 flagellar antigen, and one strain with a flagellar antigen other than H7. Optimum results were achieved when specimens were pretreated with 5% bleach and centrifugation. No false-negative results were obtained with bleach-pretreated stool samples.