title: Genomic analysis of a multidrug-resistant strain of enterohaemorrhagic Escherichia coli O157:H7 causing a family outbreak in Japan.
abstract: A family outbreak of enterohaemorrhagic Escherichia coli (EHEC) O157:H7 infection occurred in October 2003 in the Hiroshima prefecture, Japan. Four isolates of EHEC O157:H7, 03064, 03065, 03066 and 03067, were recovered from a 1-year-old daughter, mother, father and 3-year-old daughter, respectively. All EHEC O157:H7 isolates were positive for Stx1 and Stx2 Shiga toxins. Surprisingly, DNA fingerprinting profiles obtained by PFGE showed that the first isolate, 03064, had unique XbaI and BlnI profiles that differed from the other three isolates. Also, plasmid analysis results revealed that isolate 03064 contained an extra plasmid larger than the classic large plasmid of EHEC O157, pO157 (93.6 kb). This new plasmid was named pMDR157. Furthermore, isolate 03064 showed a multidrug-resistance (MDR) phenotype against streptomycin, spectinomycin, co-trimoxazole (trimethoprim/sulfamethoxazole), ampicillin and tetracycline; the other isolates were completely sensitive to these antibiotics. Molecular analysis of the MDR phenotype in this unique strain revealed the presence of a class 1 integron containing two gene cassettes: a dihydrofolate reductase type 1 gene (dfrI), which confers resistance to trimethoprim, and an aminoglycoside adenyltransferase gene (aadA1), which confers resistance to streptomycin and spectinomycin. Southern blot hybridization showed that the class 1 integron was located in the extra plasmid, pMDR157. The ampicillin resistance was found to be due to the presence of the TEM-1-type beta-lactamase gene. The MDR phenotype was transferred successfully to E. coli HB101 by conjugation, indicating that both the class 1 integron and the TEM-1 beta-lactamase were located on the conjugative transferable plasmid, pMDR157. To the authors' knowledge, this is the first report of the identification of a beta-lactamase gene in EHEC O157.
authors.affiliation: Laboratory of Food Microbiology and Hygiene, Graduate School of Biosphere Science, Hiroshima University, Higashi-Hiroshima 739-8528, Japan 2Division of Biological Science, Hiroshima City Institute of Public Health, Hiroshima 733-8650, Japan 3Division of First Microbiology, Hiroshima Prefectural Institute of Public Health and Environment, Hiroshima 734-0007, Japan.
authors.forename: Ashraf M
authors.initials: AM
authors.lastname: Ahmed
authors.affiliation: Laboratory of Food Microbiology and Hygiene, Graduate School of Biosphere Science, Hiroshima University, Higashi-Hiroshima 739-8528, Japan 2Division of Biological Science, Hiroshima City Institute of Public Health, Hiroshima 733-8650, Japan 3Division of First Microbiology, Hiroshima Prefectural Institute of Public Health and Environment, Hiroshima 734-0007, Japan.
authors.forename: Hidekazu
authors.initials: H
authors.lastname: Kawamoto
authors.affiliation: Laboratory of Food Microbiology and Hygiene, Graduate School of Biosphere Science, Hiroshima University, Higashi-Hiroshima 739-8528, Japan 2Division of Biological Science, Hiroshima City Institute of Public Health, Hiroshima 733-8650, Japan 3Division of First Microbiology, Hiroshima Prefectural Institute of Public Health and Environment, Hiroshima 734-0007, Japan.
authors.forename: Kumiko
authors.initials: K
authors.lastname: Inouye
authors.affiliation: Laboratory of Food Microbiology and Hygiene, Graduate School of Biosphere Science, Hiroshima University, Higashi-Hiroshima 739-8528, Japan 2Division of Biological Science, Hiroshima City Institute of Public Health, Hiroshima 733-8650, Japan 3Division of First Microbiology, Hiroshima Prefectural Institute of Public Health and Environment, Hiroshima 734-0007, Japan.
authors.forename: Yoshiko
authors.initials: Y
authors.lastname: Hashiwata
authors.affiliation: Laboratory of Food Microbiology and Hygiene, Graduate School of Biosphere Science, Hiroshima University, Higashi-Hiroshima 739-8528, Japan 2Division of Biological Science, Hiroshima City Institute of Public Health, Hiroshima 733-8650, Japan 3Division of First Microbiology, Hiroshima Prefectural Institute of Public Health and Environment, Hiroshima 734-0007, Japan.
authors.forename: Miyoko
authors.initials: M
authors.lastname: Sakaki
authors.affiliation: Laboratory of Food Microbiology and Hygiene, Graduate School of Biosphere Science, Hiroshima University, Higashi-Hiroshima 739-8528, Japan 2Division of Biological Science, Hiroshima City Institute of Public Health, Hiroshima 733-8650, Japan 3Division of First Microbiology, Hiroshima Prefectural Institute of Public Health and Environment, Hiroshima 734-0007, Japan.
authors.forename: Masato
authors.initials: M
authors.lastname: Seno
authors.affiliation: Laboratory of Food Microbiology and Hygiene, Graduate School of Biosphere Science, Hiroshima University, Higashi-Hiroshima 739-8528, Japan 2Division of Biological Science, Hiroshima City Institute of Public Health, Hiroshima 733-8650, Japan 3Division of First Microbiology, Hiroshima Prefectural Institute of Public Health and Environment, Hiroshima 734-0007, Japan.
authors.forename: Tadashi
authors.initials: T
authors.lastname: Shimamoto
mesh_terms: D000900:Anti-Bacterial Agents / Q000494:pharmacology; D002675:Child, Preschool; D004196:Disease Outbreaks; D024901:Drug Resistance, Multiple, Bacterial / Q000235:genetics*; D004927:Escherichia coli Infections / Q000453:epidemiology* / Q000382:microbiology; D019453:Escherichia coli O157 / Q000187:drug effects* / Q000235:genetics; D029968:Escherichia coli Proteins / Q000235:genetics / Q000378:metabolism; D005260:Female; D016680:Genome, Bacterial; D006801:Humans; D007223:Infant; D032023:Integrons / Q000235:genetics; D007564:Japan / Q000453:epidemiology; D008297:Male; D008826:Microbial Sensitivity Tests; D008969:Molecular Sequence Data; D009679:Nuclear Family; D017422:Sequence Analysis, DNA; D001618:beta-Lactamases / Q000235:genetics / Q000378:metabolism
chemical_list: D000900:Anti-Bacterial Agents; D029968:Escherichia coli Proteins; D001618:beta-Lactamases; C034413:beta-lactamase TEM-1