title: Development of a Flow Cytometry-Based Method for Rapid Detection of Escherichia coli and Shigella Spp. Using an Oligonucleotide Probe.
abstract: Standard methods to detect Escherichia coli contamination in food use the polymerase chain reaction (PCR) and agar culture plates. These methods require multiple incubation steps and take a long time to results. An improved rapid flow-cytometry based detection method was developed, using a fluorescence-labeled oligonucleotide probe specifically binding a16S rRNA sequence. The method positively detected 51 E. coli isolates as well as 4 Shigella species. All 27 non-E. coli strains tested gave negative results. Comparison of the new genetic assay with a total plate count (TPC) assay and agar plate counting indicated similar sensitivity, agreement between cytometry cell and colony counts. This method can detect a small number of E.coli cells in the presence of large numbers of other bacteria. This method can be used for rapid, economical, and stable detection of E. coli and Shigella contamination in the food industry and other contexts.
authors.affiliation: Division of Systems Biology, National Center for Toxicological Research, U.S. Food and Drug Administration, Jefferson, AR, United States of America.
authors.forename: Yong
authors.initials: Y
authors.lastname: Xue
authors.affiliation: Division of Systems Biology, National Center for Toxicological Research, U.S. Food and Drug Administration, Jefferson, AR, United States of America.
authors.forename: Jon G
authors.initials: JG
authors.lastname: Wilkes
authors.affiliation: Life Sciences Consultant, 515 W. Matthews Ave., Jonesboro, AR, United States of America.
authors.forename: Ted J
authors.initials: TJ
authors.lastname: Moskal
authors.affiliation: Division of Systems Biology, National Center for Toxicological Research, U.S. Food and Drug Administration, Jefferson, AR, United States of America.
authors.forename: Anna J
authors.initials: AJ
authors.lastname: Williams
authors.affiliation: Division of Systems Biology, National Center for Toxicological Research, U.S. Food and Drug Administration, Jefferson, AR, United States of America.
authors.forename: Willie M
authors.initials: WM
authors.lastname: Cooper
authors.affiliation: Division of Microbiology, National Center for Toxicological Research, U.S. Food and Drug Administration, Jefferson, AR, United States of America.
authors.forename: Rajesh
authors.initials: R
authors.lastname: Nayak
authors.affiliation: Division of Microbiology, National Center for Toxicological Research, U.S. Food and Drug Administration, Jefferson, AR, United States of America.
authors.forename: Fatemeh
authors.initials: F
authors.lastname: Rafii
authors.affiliation: Division of Systems Biology, National Center for Toxicological Research, U.S. Food and Drug Administration, Jefferson, AR, United States of America.
authors.forename: Dan A
authors.initials: DA
authors.lastname: Buzatu
mesh_terms: D004405:Dysentery, Bacillary / Q000382:microbiology / Q000517:prevention & control; D004927:Escherichia coli Infections / Q000382:microbiology / Q000517:prevention & control; D019453:Escherichia coli O157 / Q000235:genetics / Q000302:isolation & purification*; D005434:Flow Cytometry / Q000379:methods*; D005506:Food Contamination / Q000032:analysis*; D005517:Foodborne Diseases / Q000382:microbiology / Q000517:prevention & control; D006801:Humans; D015345:Oligonucleotide Probes / Q000235:genetics*; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D012760:Shigella / Q000235:genetics / Q000302:isolation & purification*
chemical_list: D015345:Oligonucleotide Probes; D012336:RNA, Ribosomal, 16S