In the present study, the relative sensitivity, specificity and accuracy of a real-time PCR assay for Listeria monocytogenes detection in naturally contaminated pork cuts were evaluated in comparison to the ISO 11290-1:2004 reference culture method. One hundred and sixty meat samples were collected from ten different lots over a year. The PCR method included a 24-h primary enrichment step, a DNA extraction step, and a final 5 ′ nuclease real-time PCR assay, including an internalamplification control (IAC) and targeting the hlyA gene. Based on the analysis of 480 sub-units (three sub-units for each sample), the relative sensitivity, specificity and accuracy of the real-time PCR assay were 77.3 %, 67.0 % and 73.1 %, respectively, corresponding to a Cohen ’s kappa value of 0.69 (good agreement). Considering that real samples from the worse storage scenarios were included, these results suggest the PCR method as a rapid and accurate alternative method for the quick check of L. monocytogenes in meat lots before distribution
Alessandra De Cesare, Frédérique Pasquali, Alex Lucchi, Gerardo Manfreda (2014). Relative Accuracy, Specificity and Sensitivity of a 5′ Nuclease Real-Time PCR Assay for Listeria monocytogenes Detection in Naturally Contaminated Pork Cuts. FOOD ANALYTICAL METHODS, 7, 1359-1365 [10.1007/s12161-014-9819-5].
Relative Accuracy, Specificity and Sensitivity of a 5′ Nuclease Real-Time PCR Assay for Listeria monocytogenes Detection in Naturally Contaminated Pork Cuts
DE CESARE, ALESSANDRA;PASQUALI, FREDERIQUE;LUCCHI, ALEX;MANFREDA, GERARDO
2014
Abstract
In the present study, the relative sensitivity, specificity and accuracy of a real-time PCR assay for Listeria monocytogenes detection in naturally contaminated pork cuts were evaluated in comparison to the ISO 11290-1:2004 reference culture method. One hundred and sixty meat samples were collected from ten different lots over a year. The PCR method included a 24-h primary enrichment step, a DNA extraction step, and a final 5 ′ nuclease real-time PCR assay, including an internalamplification control (IAC) and targeting the hlyA gene. Based on the analysis of 480 sub-units (three sub-units for each sample), the relative sensitivity, specificity and accuracy of the real-time PCR assay were 77.3 %, 67.0 % and 73.1 %, respectively, corresponding to a Cohen ’s kappa value of 0.69 (good agreement). Considering that real samples from the worse storage scenarios were included, these results suggest the PCR method as a rapid and accurate alternative method for the quick check of L. monocytogenes in meat lots before distributionI documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
