Molecular subtyping of Clostridium perfringens by pulsed-field gel electrophoresis to facilitate food-borne-disease outbreak investigations - PubMed (original) (raw)

Molecular subtyping of Clostridium perfringens by pulsed-field gel electrophoresis to facilitate food-borne-disease outbreak investigations

S E Maslanka et al. J Clin Microbiol. 1999 Jul.

Abstract

Clostridium perfringens is a common cause of food-borne illness. The illness is characterized by profuse diarrhea and acute abdominal pain. Since the illness is usually self-limiting, many cases are undiagnosed and/or not reported. Investigations are often pursued after an outbreak involving large numbers of people in institutions, at restaurants, or at catered meals. Serotyping has been used in the past to assist epidemiologic investigations of C. perfringens outbreaks. However, serotyping reagents are not widely available, and many isolates are often untypeable with existing reagents. We developed a pulsed-field gel electrophoresis (PFGE) method for molecular subtyping of C. perfringens isolates to aid in epidemiologic investigations of food-borne outbreaks. Six restriction endonucleases (SmaI, ApaI, FspI, MluI, KspI, and XbaI) were evaluated with a select panel of C. perfringens strains. SmaI was chosen for further studies because it produced 11 to 13 well-distributed bands of 40 to approximately 1,100 kb which provided good discrimination between isolates. Seventeen distinct patterns were obtained with 62 isolates from seven outbreak investigations or control strains. In general, multiple isolates from a single individual had indistinguishable PFGE patterns. Epidemiologically unrelated isolates (outbreak or control strains) had unique patterns; isolates from different individuals within an outbreak had similar, if not identical, patterns. PFGE identifies clonal relationships of isolates which will assist epidemiologic investigations of food-borne-disease outbreaks caused by C. perfringens.

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Figures

FIG. 1

FIG. 1

Dendrogram of PFGE subtype patterns of all C. perfringens control strains and outbreak isolates. Analysis range was 40 to 1,400 kb. CPerf1 was used as the reference standard.

FIG. 2

FIG. 2

PFGE of C. perfringens strains associated with a single outbreak (NJ 1997). Lanes 1 and 8, reference standard Cperf1; lanes 2 through 7, one isolate from each individual in the outbreak (subtype pattern L).

FIG. 3

FIG. 3

PFGE of C. perfringens associated with a single outbreak with possible multiple infecting strains (CA 1983). Lanes 1 and 10, reference standard Cperf1; lanes 2 through 4 and 7 through 9, individuals with subtype pattern A; lane 5, individual with subtype pattern G; lane 6, individual with subtype pattern N.

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