Abstract
Cronobacter (formerly known as Enterobacter sakazakii) is a genus comprising seven species regarded as opportunistic pathogens that can be found in a wide variety of environments and foods, including powdered infant formula (PIF). Cronobacter sakazakii, the major species of this genus, has been epidemiologically linked to cases of bacteremia, meningitis in neonates, and necrotizing enterocolitis, and contaminated PIF has been identified as an important source of infection. Robust and reproducible subtyping methods are required to aid in the detection and investigation, of foodborne outbreaks. In this study, a pulsed-field gel electrophoresis (PFGE) protocol was developed and validated for subtyping Cronobacter species. It was derived from an existing modified PulseNet protocol, wherein XbaI and SpeI were the primary and secondary restriction enzymes used, generating an average of 14.7 and 20.3 bands, respectively. The PFGE method developed was both reproducible and discriminatory for subtyping Cronobacter species.
Publication types
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Comparative Study
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Research Support, U.S. Gov't, P.H.S.
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Validation Study
MeSH terms
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Amplified Fragment Length Polymorphism Analysis
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Animals
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Cronobacter / classification*
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Cronobacter / genetics
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Cronobacter / isolation & purification
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Cronobacter / metabolism
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Cronobacter sakazakii / classification
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Cronobacter sakazakii / genetics
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Cronobacter sakazakii / isolation & purification
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Cronobacter sakazakii / metabolism
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DNA Restriction Enzymes
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DNA, Bacterial / chemistry
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DNA, Bacterial / genetics
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DNA, Bacterial / metabolism
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Electrophoresis, Gel, Pulsed-Field
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Enterobacteriaceae Infections / diagnosis
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Enterobacteriaceae Infections / microbiology
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Food, Preserved / microbiology
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Foodborne Diseases / diagnosis
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Foodborne Diseases / microbiology
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Humans
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Milk / microbiology
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Molecular Typing / methods*
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Reproducibility of Results
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Vibrio cholerae / classification
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Vibrio cholerae / genetics
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Vibrio cholerae / isolation & purification
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Vibrio cholerae / metabolism
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Yersinia pestis / classification
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Yersinia pestis / genetics
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Yersinia pestis / isolation & purification
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Yersinia pestis / metabolism
Substances
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DNA, Bacterial
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DNA Restriction Enzymes