Document Detail


Development of a real-time PCR assay for detection and quantification of enterotoxigenic members of Bacillus cereus group in food samples.
MedLine Citation:
PMID:  19665814     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
A highly sensitive real-time PCR (qPCR) procedure, targeting the phosphatidylcholine-specific phospholipase C gene (pc-plc), was developed for specific detection and quantification of strains belonging to Bacillus cereus group. The target region was selected based on the enterotoxigenic profiles of 75 Bacillus strains. The inclusivity and exclusivity of the RTi-PCR assay were assessed with 59 isolates of the B. cereus group, 16 other Bacillus spp., and 4 non-Bacillus strains. The assay was also used to construct calibration curves for different food matrices, and it had a wide quantification range of 6 log units using both serial dilutions of purified DNA and calibrated cell suspensions of B. cereus CECT 148(T). The detection limit for B. cereus in artificially contaminated liquid egg and reconstituted infant formula was about 3CFU per reaction or 60CFU/ml of food, with a relative accuracy of 86.27% to 116.12% in artificially contaminated liquid egg. Naturally contaminated food samples were tested for the presence of B. cereus with the standard method, a conventional PCR and the new developed RTi-PCR assay. Results showed that the new developed RTi-PCR assay is very suitable for detection and quantification of strains of B. cereus group in food samples without an enrichment step.
Authors:
J F Martínez-Blanch; G Sánchez; E Garay; R Aznar
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't     Date:  2009-07-19
Journal Detail:
Title:  International journal of food microbiology     Volume:  135     ISSN:  1879-3460     ISO Abbreviation:  Int. J. Food Microbiol.     Publication Date:  2009 Sep 
Date Detail:
Created Date:  2009-08-26     Completed Date:  2009-11-17     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  8412849     Medline TA:  Int J Food Microbiol     Country:  Netherlands    
Other Details:
Languages:  eng     Pagination:  15-21     Citation Subset:  IM    
Affiliation:
University of Valencia, Spain.
Data Bank Information
Bank Name/Acc. No.:
GENBANK/FJ810833;  FJ810834;  FJ810835;  FJ810836;  FJ810837;  FJ810838;  FJ810839;  FJ810840;  FJ810841;  FJ810842;  FJ810843;  FJ810844;  FJ810845;  FJ810846;  FJ810847;  FJ810848
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MeSH Terms
Descriptor/Qualifier:
Bacillus cereus / classification,  genetics,  isolation & purification*,  metabolism
Calibration
Colony Count, Microbial
DNA, Bacterial / chemistry
Eggs / microbiology
Enterotoxins / biosynthesis*,  isolation & purification
Food Contamination / analysis*
Food Microbiology
Humans
Infant
Infant Formula
Infant, Newborn
Molecular Sequence Data
Polymerase Chain Reaction / methods*
Reproducibility of Results
Sensitivity and Specificity
Sequence Analysis, DNA
Species Specificity
Chemical
Reg. No./Substance:
0/DNA, Bacterial; 0/Enterotoxins

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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