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Triplex PCR-based detection of enterotoxigenic Bacillus cereus ATCC 14579 in nonfat dry milk.
MedLine Citation:
PMID:  20586065     Owner:  NLM     Status:  In-Data-Review    
Abstract/OtherAbstract:
Although many strains of Bacillaceae are considered nonpathogenic, Bacillus cereus is recognized worldwide as a bacterial pathogen in a variety of foods. The ability of B. cereus to cause gastroenteritis following ingestion of contaminated food is due to the production of enterotoxins. The ubiquity of this genus makes it a persistent problem for quality assurance in food processing environments. The primary objective of this study was to develop and apply a multiplex real-time PCR-based assay for rapid and sensitive detection of enterotoxigenic B. cereus. Template DNA was separately extracted from tryptic soy broth (TSB)-grown and 2.5% Nonfat Dry Milk (NFDM)-grown B. cereus using a commercial system. Three enterotoxin gene fragments (hblC, nheA, and hblA) were simultaneously amplified in real-time followed by melting curve analysis to confirm amplicon identity. Resolution of melting curves (characteristic T(m) ) was achieved for each amplicon (hblC = 74.5 °C; nheA = 78 °C; and hblA = 85.5 °C in TSB and 84 °C in NFDM) with an assay sensitivities of 10(1) CFU/ml for both TSB and NFDM-grown B. cereus compared to 10(4) CFU/ml in either matrix using gel electrophoresis. The results demonstrate the potential sensitivity of real-time bacterial detection methods in a heterogenous food matrix using real-time PCR. (© 2011 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim).
Authors:
Kiev S Gracias; John L McKillip
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Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Journal of basic microbiology     Volume:  51     ISSN:  1521-4028     ISO Abbreviation:  J. Basic Microbiol.     Publication Date:  2011 Apr 
Date Detail:
Created Date:  2011-04-14     Completed Date:  -     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  8503885     Medline TA:  J Basic Microbiol     Country:  Germany    
Other Details:
Languages:  eng     Pagination:  147-52     Citation Subset:  IM    
Copyright Information:
Copyright © 2011 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.
Affiliation:
Department of Arts and Sciences, Oakland City University, Oakland City, IN, USA.
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