Assessment of food safety using a new real-time PCR assay for detection and quantification of virulence factors of enterococci in food samples.
OBJECTIVE
Development of Taqman MGB real-time PCR (q-PCR) assays for the quantitative detection of virulence factor genes in pure culture and food samples with regard to food safety assessment.


RESULTS
New Taqman primers and probes were designed for the ace, esp and gelE genes based on the determinants of virulence profiles of enterococcal strains from GenBank. The high specificity and accuracy of the Taqman probe assay was confirmed. The limit of detection for the different virulence genes was 102CFU ml-1or CFU g-1for pure culture and meat samples, and 103CFU g-1for cheese samples.


CONCLUSIONS
This method provides the specific and rapid detection and quantification of ace, esp and gelE genes compared to conventional PCR assays, thus allowing the rapid and direct safety assessment of Enterococcus genus in food samples.


CONCLUSIONS
This study presents efficient methods that can be used directly on food products for the rapid quantification and tracing of virulence genes, regarding food safety assessment. Moreover, this is the first study to quantify these virulence factors using a specific Taqman q-PCR assay in food samples.
Department of Analytical Chemistry, Nutrition and Food Science, School of Veterinary Sciences, University of Santiago de Compostela, Lugo, Spain.|Dairy Department, Faculty of Agriculture, Suez Canal University, Ismailia, Egypt.
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M
Abouelnaga
Department of Analytical Chemistry, Nutrition and Food Science, School of Veterinary Sciences, University of Santiago de Compostela, Lugo, Spain.
A
A
Lamas
Department of Analytical Chemistry, Nutrition and Food Science, School of Veterinary Sciences, University of Santiago de Compostela, Lugo, Spain.
M
M
Guarddon
Dairy Department, Faculty of Agriculture, Suez Canal University, Ismailia, Egypt.
M
M
Osman
Department of Analytical Chemistry, Nutrition and Food Science, School of Veterinary Sciences, University of Santiago de Compostela, Lugo, Spain.
J M
JM
Miranda
Department of Analytical Chemistry, Nutrition and Food Science, School of Veterinary Sciences, University of Santiago de Compostela, Lugo, Spain.
A
A
Cepeda
Department of Analytical Chemistry, Nutrition and Food Science, School of Veterinary Sciences, University of Santiago de Compostela, Lugo, Spain.
C M
CM
Franco
D017931:DNA Primers; D016983:Enterococcus / Q000235:genetics / Q000302:isolation & purification* / Q000472:pathogenicity; D005516:Food Microbiology; D059022:Food Safety; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D037521:Virulence Factors / Q000235:genetics
D017931:DNA Primers; D037521:Virulence Factors
enterococci; food safety; quantification; real-time PCR; virulence factors