Document Detail


Validation of iQ-Check E. coli O157:H7 real-time PCR test kit for detection of Escherichia coli O157:H7 in selected foods.
MedLine Citation:
PMID:  19714978     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
iQ-Check E. coli O157:H7 (Bio-Rad Laboratories, Hercules, CA) is a real-time PCR kit for detection of E. coli O157:H7 from selected foods. Specific fluorescent oligonucleotide probes are used to detect target DNA during the amplification, by hybridizing to the amplicons. These fluorescent probes are linked to a fluorophore which fluoresces only when hybridized to the target sequence. Three foods (ground beef, apple cider, fresh spinach) were selected to compare the performance of iQ-Check E. coli O157:H7 to the U.S. Department of Agriculture-Food Safety and Inspection Service Microbiology Laboratory Guidebook (MLG) reference method for ground beef and the U.S. Food and Drug Administration/Bacteriological Analytical Manual reference method for apple cider and fresh spinach. Three protocols were tested in this study: a shortened 8 h primary enrichment in buffered peptone water (BPW), a 24 h enrichment in BPW, and an enrichment in appropriate reference method enrichment broth. The iQ-Check E. coli O157:H7 method was able to identify more true/confirmed positive samples than the reference method. Inclusivity and exclusivity rates of the method were 100%. iQ-Check E. coli O157:H7 performed as expected when minor procedural variations were introduced, validating the ruggedness of the method. There was no significant difference observed in performance over the shelf life of the kit.
Authors:
Wendy F Lauer; Sylvie Tymciu; Caroline D Sidi; Pierre Sonigo
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Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Journal of AOAC International     Volume:  92     ISSN:  1060-3271     ISO Abbreviation:  J AOAC Int     Publication Date:    2009 Jul-Aug
Date Detail:
Created Date:  2009-08-31     Completed Date:  2009-09-24     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  9215446     Medline TA:  J AOAC Int     Country:  United States    
Other Details:
Languages:  eng     Pagination:  1095-104     Citation Subset:  IM    
Affiliation:
Bio-Rad Laboratories, 2000 Alfred Nobel Dr, Hercules, CA 94564, USA. wendy_lauer@bio-rad.com
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MeSH Terms
Descriptor/Qualifier:
Animals
Cattle
DNA, Bacterial / analysis,  genetics
Escherichia coli O157 / chemistry*,  genetics
Food Microbiology*
Malus / microbiology
Meat / microbiology
Reference Standards
Reproducibility of Results
Reverse Transcriptase Polymerase Chain Reaction
Spinacia oleracea / microbiology
Chemical
Reg. No./Substance:
0/DNA, Bacterial

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


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