Document Detail


A scorpion probe-based real-time PCR assay for detection of E. coli O157:H7 in dairy products.
MedLine Citation:
PMID:  19272008     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Methods for rapidly detecting Escherichia coli O157:H7, a high risk foodborne pathogen of considerable public health concern, are extremely important for monitoring of food safety. Conventional detection such as microbiological, biochemical, and immunological methods, including polymerase chain reactions (PCR), are time consuming and only qualitative. In this study, a rapid real-time PCR (RTi-PCR) method based on Scorpion probe technology targeting the eae gene of E. coli O157:H7 was developed and applied using spiked and naturally contaminated products. The assay was highly specific and did not show any cross-reactivity with any of the 15 nontargeted bacterial cultures. The assay was sensitive enough to detect 2 log colony-forming units (CFU)/mL of the target pathogen in pure broth culture and 3 log CFU/mL in spiked skim milk. When the assay was applied with 60 market dairy food samples, one sample each of raw milk, paneer, and ice cream, was found to be positive for E. coli O157:H7 with respective RTi-PCR counts of 6.7, 6.2, and 5.9 log CFU. The present study demonstrates the application of Scorpion probe technology for the detection of foodborne pathogens and reports the presence of E. coli O157:H7 in paneer, a very popular indigenous dairy product (soft cheese) in India, for the first time.
Authors:
Jitender Singh; Virender K Batish; Sunita Grover
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Foodborne pathogens and disease     Volume:  6     ISSN:  1556-7125     ISO Abbreviation:  Foodborne Pathog. Dis.     Publication Date:  2009 Apr 
Date Detail:
Created Date:  2009-04-03     Completed Date:  2009-07-30     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  101120121     Medline TA:  Foodborne Pathog Dis     Country:  United States    
Other Details:
Languages:  eng     Pagination:  395-400     Citation Subset:  IM    
Affiliation:
National Dairy Research Institute, Karnal, India.
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MeSH Terms
Descriptor/Qualifier:
Adhesins, Bacterial / genetics
Animals
Cheese / microbiology
DNA, Bacterial / analysis
Dairy Products / microbiology*
Escherichia coli Infections / microbiology,  prevention & control,  transmission
Escherichia coli O157 / genetics,  isolation & purification*
Escherichia coli Proteins / genetics
Humans
Ice Cream / microbiology
Milk / microbiology
Polymerase Chain Reaction / methods*
Sensitivity and Specificity
Chemical
Reg. No./Substance:
0/Adhesins, Bacterial; 0/DNA, Bacterial; 0/Escherichia coli Proteins; 147094-99-3/eaeA protein, E coli

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


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