| Rapid separation and concentration of food-borne pathogens in food samples prior to quantification by viable-cell counting and real-time PCR. | |
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MedLine Citation:
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PMID: 17056684 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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Buoyant density gradient centrifugation has been used to separate bacteria from complex food matrices, as well as to remove compounds that inhibit rapid detection methods, such as PCR, and to prevent false-positive results due to DNA originating from dead cells. Applying a principle of buoyant density gradient centrifugation, we developed a method for rapid separation and concentration following filtration and low- and high-speed centrifugation, as well as flotation and sedimentation buoyant density centrifugation, for 12 food-borne pathogens (Salmonella enterica, Escherichia coli, Yersinia enterocolitica, Campylobacter jejuni, Vibrio cholerae O139, Vibrio parahaemolyticus O3K6, Vibrio vulnificus, Providencia alcalifaciens, Aeromonas hydrophila, Bacillus cereus, Staphylococcus aureus, and Clostridium perfringens) in 13 different food homogenates. This method can be used prior to real-time quantitative PCR (RTi-qPCR) and viable-cell counting. Using this combined method, the target organisms in the food samples theoretically could be concentrated 250-fold and detected at cell concentrations as low as 10(1) to 10(3) CFU/g using the RTi-qPCR assay, and amounts as small as 10(0) to 10(1) CFU/g could be isolated using plate counting. The combined separation and concentration methods and RTi-qPCR confirmed within 3 h the presence of 10(1) to 10(2) CFU/g of Salmonella and C. jejuni directly in naturally contaminated chicken and the presence of S. aureus directly in remaining food items in a poisoning outbreak. These results illustrated the feasibility of using these assays for rapid inspection of bacterial food contamination during a real-world outbreak. |
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Authors:
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Hiroshi Fukushima; Kazunori Katsube; Yukiko Hata; Ryoko Kishi; Satomi Fujiwara |
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Publication Detail:
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Type: Evaluation Studies; Journal Article Date: 2006-10-20 |
Journal Detail:
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Title: Applied and environmental microbiology Volume: 73 ISSN: 0099-2240 ISO Abbreviation: Appl. Environ. Microbiol. Publication Date: 2007 Jan |
Date Detail:
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Created Date: 2006-12-27 Completed Date: 2007-05-25 Revised Date: 2009-11-19 |
Medline Journal Info:
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Nlm Unique ID: 7605801 Medline TA: Appl Environ Microbiol Country: United States |
Other Details:
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Languages: eng Pagination: 92-100 Citation Subset: IM |
Affiliation:
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Shimane Prefectural Institute of Public Health and Environmental Science, 582 Nishihamasada, Matsue, Shimane 690-0122, Japan. fukushima-hiroshi@pref.shimane.lg.jp |
Export Citation:
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| MeSH Terms | |
Descriptor/Qualifier:
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Animals Campylobacter jejuni / isolation & purification Cattle Centrifugation, Density Gradient / methods Chickens / microbiology Colony Count, Microbial DNA, Bacterial / analysis, isolation & purification Dairy Products / microbiology Filtration Food Contamination / analysis Food Microbiology* Foodborne Diseases / microbiology* Gram-Negative Bacteria / isolation & purification Gram-Positive Bacteria / isolation & purification Humans Meat / microbiology Perciformes / microbiology Polymerase Chain Reaction Salmonella enterica / classification, isolation & purification Time Factors |
| Chemical | |
Reg. No./Substance:
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0/DNA, Bacterial |
| Comments/Corrections | |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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