title: Development of a multiplex PCR-based rapid typing method for enterohemorrhagic Escherichia coli O157 strains.
abstract: Enterohemorrhagic Escherichia coli O157 (EHEC O157) is a food-borne pathogen that has raised worldwide public health concern. The development of simple and rapid strain-typing methods is crucial for the rapid detection and surveillance of EHEC O157 outbreaks. In the present study, we developed a multiplex PCR-based strain-typing method for EHEC O157, which is based on the variability in genomic location of IS629 among EHEC O157 strains. This method is very simple, in that the procedures are completed within 2 h, the analysis can be performed without the need for special equipment or techniques (requiring only conventional PCR and agarose gel electrophoresis systems), the results can easily be transformed into digital data, and the genes for the major virulence markers of EHEC O157 (the stx(1), stx(2), and eae genes) can be detected simultaneously. Using this method, 201 EHEC O157 strains showing different XbaI digestion patterns in pulsed-field gel electrophoresis (PFGE) analysis were classified into 127 types, and outbreak-related strains showed identical or highly similar banding patterns. Although this method is less discriminatory than PFGE, it may be useful as a primary screening tool for EHEC O157 outbreaks.
authors.affiliation: Division of Microbiology, Department of Infectious Diseases, Faculty of Medicine, University of Miyazaki, Miyazaki 889-1692, Japan.
authors.forename: Tadasuke
authors.initials: T
authors.lastname: Ooka
authors.forename: Jun
authors.initials: J
authors.lastname: Terajima
authors.forename: Masahiro
authors.initials: M
authors.lastname: Kusumoto
authors.forename: Atsushi
authors.initials: A
authors.lastname: Iguchi
authors.forename: Ken
authors.initials: K
authors.lastname: Kurokawa
authors.forename: Yoshitoshi
authors.initials: Y
authors.lastname: Ogura
authors.forename: Md
authors.initials: M
authors.lastname: Asadulghani
authors.forename: Keisuke
authors.initials: K
authors.lastname: Nakayama
authors.forename: Kazunori
authors.initials: K
authors.lastname: Murase
authors.forename: Makoto
authors.initials: M
authors.lastname: Ohnishi
authors.forename: Sunao
authors.initials: S
authors.lastname: Iyoda
authors.forename: Haruo
authors.initials: H
authors.lastname: Watanabe
authors.forename: Tetsuya
authors.initials: T
authors.lastname: Hayashi
mesh_terms: D015373:Bacterial Typing Techniques / Q000379:methods*; D016000:Cluster Analysis; D016172:DNA Fingerprinting / Q000379:methods; D017931:DNA Primers / Q000235:genetics; D004251:DNA Transposable Elements; D004269:DNA, Bacterial / Q000235:genetics*; D004927:Escherichia coli Infections / Q000382:microbiology*; D019453:Escherichia coli O157 / Q000145:classification* / Q000235:genetics*; D005838:Genotype; D006801:Humans; D017720:Molecular Epidemiology / Q000379:methods; D016133:Polymerase Chain Reaction / Q000379:methods*; D011110:Polymorphism, Genetic; D037521:Virulence Factors / Q000235:genetics
chemical_list: D017931:DNA Primers; D004251:DNA Transposable Elements; D004269:DNA, Bacterial; D037521:Virulence Factors