title: Applicability of DiversiLab repetitive sequence-based PCR method in epidemiological typing of enterohemorrhagic Escherichia coli (EHEC).
abstract: Enterohemorrhagic Escherichia coli (EHEC) causes diarrhea, often with severe complications. Rapid and discriminatory typing of EHEC using advanced molecular methods is needed for determination of the genetic relatedness of clones responsible for foodborne outbreaks and for finding out the transmission sources of the outbreaks. This study evaluated the potential of DiversiLab, a semiautomated repetitive sequence-based polymerase chain reaction method for the genotyping of EHEC strains. A set of 52 EHEC strains belonging to 15 O:H serotypes was clustered into 10 DiversiLab groups. All of the O157 strains and one O55 strain were classified into the same group based on a 90% similarity threshold. The other serotypes were classified to their own DiversiLab group, with the exception of one R:H(-) strain that was grouped with O5:H(-) strains. In addition, O26 and O111 strains were grouped together but ultimately subdivided according to their O-serotypes based on a 95% similarity threshold. The O104 strain, which was associated with a major outbreak of hemolytic uremic syndrome in Germany in May 2011, was also classified independently. The DiversiLab performed well in identifying isolates, but the discriminatory power of the repetitive sequence-based polymerase chain reaction method was lower than that of pulsed-field gel electrophoresis. Analysis of 15 enteropathogenic E. coli (EPEC) strains revealed that some EPEC strains clustered together with EHEC strains. Therefore, the DiversiLab system cannot be used to discriminate between these pathogroups. In conclusion, DiversiLab is a rapid and easy system for the primary exclusion of unrelated EHEC strains based on their serotypes, but more discriminatory methods, such as pulsed-field gel electrophoresis, are needed for accurate typing of the EHEC strains.
authors.affiliation: Bacteriology Unit, Department of Infectious Disease Surveillance and Control, National Institute for Health and Welfare THL, Helsinki, Finland.
authors.forename: Susanna
authors.initials: S
authors.lastname: Lukinmaa-Åberg
authors.forename: Jenni
authors.initials: J
authors.lastname: Horsma
authors.forename: Tanja
authors.initials: T
authors.lastname: Pasanen
authors.forename: Sointu
authors.initials: S
authors.lastname: Mero
authors.forename: Laura
authors.initials: L
authors.lastname: Aulu
authors.forename: Martti
authors.initials: M
authors.lastname: Vaara
authors.forename: Anja
authors.initials: A
authors.lastname: Siitonen
authors.forename: Jenni
authors.initials: J
authors.lastname: Antikainen
mesh_terms: D015373:Bacterial Typing Techniques / Q000379:methods*; D016000:Cluster Analysis; D016172:DNA Fingerprinting; D004269:DNA, Bacterial / Q000235:genetics; D004196:Disease Outbreaks; D016521:Electrophoresis, Gel, Pulsed-Field; D054324:Enterohemorrhagic Escherichia coli / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification; D004927:Escherichia coli Infections / Q000453:epidemiology / Q000382:microbiology*; D019453:Escherichia coli O157 / Q000145:classification / Q000235:genetics / Q000302:isolation & purification; D005517:Foodborne Diseases / Q000453:epidemiology / Q000382:microbiology; D005838:Genotype; D005858:Germany / Q000453:epidemiology; D006463:Hemolytic-Uremic Syndrome / Q000453:epidemiology / Q000382:microbiology*; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods; D012091:Repetitive Sequences, Nucleic Acid / Q000235:genetics*; D015203:Reproducibility of Results; D013997:Time Factors
chemical_list: D004269:DNA, Bacterial