Novel Real-Time PCR Method To Detect Escherichia coli O157:H7 in Raw Milk Cheese and Raw Ground Meat
Source: Journal of Food Protection®, Volume 75, Number 8, August 2012 , pp. 1373-1381(9)
Abstract:Raw milk, raw milk cheeses, and raw ground meat have been implicated in Escherichia coli O157:H7 outbreaks. Developing methods to detect these bacteria in raw milk and meat products is a major challenge for food safety. The aim of our study was to develop a real-time PCR assay to detect E. coli O157:H7 in raw milk cheeses and raw ground meat. Well-known primers targeting a mutation at position +93 of the uidA gene in E. coli O157:H7 were chosen, and a specific TaqMan-minor groove binder probe was designed. This probe targets another mutation, at position +191 of the uidA gene in E. coli O157:H7. The first step in the study was to evaluate the specificity of this probe with 156 different O157:H7/NM strains and 48 non-O157:H7/NM strains of E. coli. The sensitivity of the method was evaluated by pre- and postinoculation of cheeses and meat enrichments with different E. coli O157:H7 strains. All the E. coli O157:H7 isolates tested were positive, and none of the other bacteria were detected. Our results indicate that this method is sensitive enough to detect 102 E. coli O157:H7 isolates per ml of cheese or meat enrichment broth (24 h at 41.5° C) and is more sensitive than the International Organization for Standardization reference method. We can conclude that this new real-time PCR protocol is a useful tool for rapid, specific, and sensitive detection of E. coli O157:H7 in raw milk and raw ground meat products.
Document Type: Research article
Affiliations: 1: Unité de Recherche CALITYSS/Equipe EMSA, VetAgro Sup, Université de Lyon, 1 avenue Claude Bourgelat, 69290 Marcy l'Etoile, France., Email: firstname.lastname@example.org 2: Unité de Recherche CALITYSS/Equipe EMSA, Laboratoire LMAP/Laboratoire National de Référence STEC, VetAgro Sup, Université de Lyon, 1 avenue Claude Bourgelat, 69290 Marcy l'Etoile, France 3: Unité de Recherche CALITYSS/Equipe EMSA, VetAgro Sup, Université de Lyon, 1 avenue Claude Bourgelat, 69290 Marcy l'Etoile, France
Publication date: 2012-08-01
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