Development of a multiplex PCR-based rapid typing method for enterohemorrhagic Escherichia coli O157 strains.
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.
Ooka
Tadasuke
T
Division of Microbiology, Department of Infectious Diseases, Faculty of Medicine, University of Miyazaki, Miyazaki 889-1692, Japan.
Terajima
Jun
J
Kusumoto
Masahiro
M
Iguchi
Atsushi
A
Kurokawa
Ken
K
Ogura
Yoshitoshi
Y
Asadulghani
Md
M
Nakayama
Keisuke
K
Murase
Kazunori
K
Ohnishi
Makoto
M
Iyoda
Sunao
S
Watanabe
Haruo
H
Hayashi
Tetsuya
T
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
D017931:DNA Primers; D004251:DNA Transposable Elements; D004269:DNA, Bacterial; D037521:Virulence Factors