The Identification and Characterization ofClostridium perfringensby Real-Time PCR, Location of Enterotoxin Gene, and Heat Resistance
| Autor: | Michael W. Peck, Kathie Grant, Sarah Kenyon, Charles Ohai, Robin Halford-Maw, Jim McLauchlin, Ijeoma Nwafor, June Plowman |
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| Rok vydání: | 2008 |
| Předmět: |
DNA
Bacterial Hot Temperature Meat Genotype Clostridium perfringens Enterotoxin Biology medicine.disease_cause Polymerase Chain Reaction Applied Microbiology and Biotechnology Microbiology law.invention Foodborne Diseases Enterotoxins Feces law medicine Humans Food microbiology Gene Polymerase chain reaction Spores Bacterial Microbial Viability Chromosomes Bacterial Virology Diarrhea Real-time polymerase chain reaction Genes Bacterial Clostridium Infections Food Microbiology Animal Science and Zoology medicine.symptom Genome Bacterial Plasmids Food Science |
| Zdroj: | Foodborne Pathogens and Disease. 5:629-639 |
| ISSN: | 1556-7125 1535-3141 |
| Popis: | Clostridium perfringens carrying the enterotoxin gene is an important cause of both foodborne and non-foodborne diarrheal disease. Rapid identification of isolates carrying the enterotoxin gene is invaluable for outbreak investigation whilst information on the genomic location of the enterotoxin (cpe) gene can improve our understanding of disease transmission. This paper describes the validation of a real-time polymerase chain reaction (PCR) assay for the identification of C. perfringens and assessment of the potential to cause diarrhea, together with an investigation into the genomic location of the cpe genes in isolates from confirmed incidents of C. perfringens diarrhea. The real-time assay was shown to be specific for the identification of 253 C. perfringens cultures and gave results concordant with those from motility nitrate and lactose gelatine media, the Nagler reaction, and a conventional block-based PCR assay. The cpe gene was detected in 223 of 253 C. perfringens cultures isolated in association with human gastrointestinal disease. A subset of cpe-positive C. perfringens isolates associated with separate incidents of diarrheal disease were investigated further for plasmid or chromosomal location of the cpe gene using a multiplex PCR assay. The cpe gene was plasmid encoded in two isolates from cases of sporadic diarrhea and six isolates from cases of food poisoning. The cpe gene from the remaining 11 isolates from different food poisoning outbreaks was found to be chromosomally encoded. One of the C. perfringens strains with a plasmid encoded cpe gene formed spores of high heat resistance and five formed spores that were sensitive to heating. Eight of the isolates with a chromosomal cpe gene formed heat-resistant spores, and two formed spores with an intermediate heat resistance. |
| Databáze: | OpenAIRE |
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