European validation of a real-time PCR-based method for detection of Listeria monocytogenes in soft cheese

Monica Virginia  Gianfranceschi; David  Rodriguez-Lazaro; Marta Hernandez; Patricia González-García; Damiano  Comin; Antonietta  Gattuso; Elisabetta  Delibato; Michele  Sonnessa; Frederique  Pasquali; Vincenza  Prencipe; Dziuginta Jakociune; John Elmerdahl Olsen

doi:10.1016/j.ijfoodmicro.2013.12.021

European validation of a real-time PCR-based method for detection of Listeria monocytogenes in soft cheese

Monica Virginia Gianfranceschi, David Rodriguez-Lazaro, Marta Hernandez, Patricia González-García, Damiano Comin, Antonietta Gattuso, Elisabetta Delibato, Michele Sonnessa, Frederique Pasquali, Vincenza Prencipe, Dziuginta Jakociune, John Elmerdahl Olsen

30 Citationer (Scopus)

Abstract

The classical microbiological method for detection of Listeria monocytogenes requires around 7. days for final confirmation, and due to perishable nature of RTE food products, there is a clear need for an alternative methodology for detection of this pathogen. This study presents an international (at European level) ISO 16140-based validation trial of a non-proprietary real-time PCR-based methodology that can generate final results in the following day of the analysis. This methodology is based on an ISO compatible enrichment coupled to a bacterial DNA extraction and a consolidated real-time PCR assay. Twelve laboratories from six European countries participated in this trial, and soft cheese was selected as food model since it can represent a difficult matrix for the bacterial DNA extraction and real-time PCR amplification. The limit of detection observed was down to 10. CFU per 25 of sample, showing excellent concordance and accordance values between samples and laboratories (>. 75%). In addition, excellent values were obtained for relative accuracy, specificity and sensitivity (82.75%, 96.70% and 97.62%, respectively) when the results obtained for the real-time PCR-based methods were compared to those of the ISO 11290-1 standard method. An interesting observation was that the L. monocytogenes detection by the real-time PCR method was less affected in the presence of Listeria innocua in the contaminated samples, proving therefore to be more reliable than the reference method. The results of this international trial demonstrate that the evaluated real-time PCR-based method represents an excellent alterative to the ISO standard since it shows a higher performance as well as reduce the extent of the analytical process, and can be easily implemented routinely by the competent authorities and food industry laboratories.

Originalsprog	Engelsk
Tidsskrift	International Journal of Food Microbiology
Vol/bind	184
Sider (fra-til)	128-133
Antal sider	6
ISSN	0168-1605
DOI	https://doi.org/10.1016/j.ijfoodmicro.2013.12.021
Status	Udgivet - 1 aug. 2014

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10.1016/j.ijfoodmicro.2013.12.021

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Gianfranceschi, M. V., Rodriguez-Lazaro, D., Hernandez, M., González-García, P., Comin, D., Gattuso, A., Delibato, E., Sonnessa, M., Pasquali, F., Prencipe, V., Jakociune, D., & Olsen, J. E. (2014). European validation of a real-time PCR-based method for detection of Listeria monocytogenes in soft cheese. International Journal of Food Microbiology, 184, 128-133. https://doi.org/10.1016/j.ijfoodmicro.2013.12.021

Gianfranceschi, MV, Rodriguez-Lazaro, D, Hernandez, M, González-García, P, Comin, D, Gattuso, A, Delibato, E, Sonnessa, M, Pasquali, F, Prencipe, V, Jakociune, D & Olsen, JE 2014, 'European validation of a real-time PCR-based method for detection of Listeria monocytogenes in soft cheese', International Journal of Food Microbiology, bind 184, s. 128-133. https://doi.org/10.1016/j.ijfoodmicro.2013.12.021

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title = "European validation of a real-time PCR-based method for detection of Listeria monocytogenes in soft cheese",

abstract = "The classical microbiological method for detection of Listeria monocytogenes requires around 7. days for final confirmation, and due to perishable nature of RTE food products, there is a clear need for an alternative methodology for detection of this pathogen. This study presents an international (at European level) ISO 16140-based validation trial of a non-proprietary real-time PCR-based methodology that can generate final results in the following day of the analysis. This methodology is based on an ISO compatible enrichment coupled to a bacterial DNA extraction and a consolidated real-time PCR assay. Twelve laboratories from six European countries participated in this trial, and soft cheese was selected as food model since it can represent a difficult matrix for the bacterial DNA extraction and real-time PCR amplification. The limit of detection observed was down to 10. CFU per 25 of sample, showing excellent concordance and accordance values between samples and laboratories (>. 75%). In addition, excellent values were obtained for relative accuracy, specificity and sensitivity (82.75%, 96.70% and 97.62%, respectively) when the results obtained for the real-time PCR-based methods were compared to those of the ISO 11290-1 standard method. An interesting observation was that the L. monocytogenes detection by the real-time PCR method was less affected in the presence of Listeria innocua in the contaminated samples, proving therefore to be more reliable than the reference method. The results of this international trial demonstrate that the evaluated real-time PCR-based method represents an excellent alterative to the ISO standard since it shows a higher performance as well as reduce the extent of the analytical process, and can be easily implemented routinely by the competent authorities and food industry laboratories.",

author = "Gianfranceschi, {Monica Virginia} and David Rodriguez-Lazaro and Marta Hernandez and Patricia Gonz{\'a}lez-Garc{\'i}a and Damiano Comin and Antonietta Gattuso and Elisabetta Delibato and Michele Sonnessa and Frederique Pasquali and Vincenza Prencipe and Dziuginta Jakociune and Olsen, {John Elmerdahl}",

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AU - Gianfranceschi, Monica Virginia

AU - Rodriguez-Lazaro, David

AU - Hernandez, Marta

AU - González-García, Patricia

AU - Comin, Damiano

AU - Gattuso, Antonietta

AU - Delibato, Elisabetta

AU - Sonnessa, Michele

AU - Pasquali, Frederique

AU - Prencipe, Vincenza

AU - Jakociune, Dziuginta

AU - Olsen, John Elmerdahl

PY - 2014/8/1

Y1 - 2014/8/1

N2 - The classical microbiological method for detection of Listeria monocytogenes requires around 7. days for final confirmation, and due to perishable nature of RTE food products, there is a clear need for an alternative methodology for detection of this pathogen. This study presents an international (at European level) ISO 16140-based validation trial of a non-proprietary real-time PCR-based methodology that can generate final results in the following day of the analysis. This methodology is based on an ISO compatible enrichment coupled to a bacterial DNA extraction and a consolidated real-time PCR assay. Twelve laboratories from six European countries participated in this trial, and soft cheese was selected as food model since it can represent a difficult matrix for the bacterial DNA extraction and real-time PCR amplification. The limit of detection observed was down to 10. CFU per 25 of sample, showing excellent concordance and accordance values between samples and laboratories (>. 75%). In addition, excellent values were obtained for relative accuracy, specificity and sensitivity (82.75%, 96.70% and 97.62%, respectively) when the results obtained for the real-time PCR-based methods were compared to those of the ISO 11290-1 standard method. An interesting observation was that the L. monocytogenes detection by the real-time PCR method was less affected in the presence of Listeria innocua in the contaminated samples, proving therefore to be more reliable than the reference method. The results of this international trial demonstrate that the evaluated real-time PCR-based method represents an excellent alterative to the ISO standard since it shows a higher performance as well as reduce the extent of the analytical process, and can be easily implemented routinely by the competent authorities and food industry laboratories.

AB - The classical microbiological method for detection of Listeria monocytogenes requires around 7. days for final confirmation, and due to perishable nature of RTE food products, there is a clear need for an alternative methodology for detection of this pathogen. This study presents an international (at European level) ISO 16140-based validation trial of a non-proprietary real-time PCR-based methodology that can generate final results in the following day of the analysis. This methodology is based on an ISO compatible enrichment coupled to a bacterial DNA extraction and a consolidated real-time PCR assay. Twelve laboratories from six European countries participated in this trial, and soft cheese was selected as food model since it can represent a difficult matrix for the bacterial DNA extraction and real-time PCR amplification. The limit of detection observed was down to 10. CFU per 25 of sample, showing excellent concordance and accordance values between samples and laboratories (>. 75%). In addition, excellent values were obtained for relative accuracy, specificity and sensitivity (82.75%, 96.70% and 97.62%, respectively) when the results obtained for the real-time PCR-based methods were compared to those of the ISO 11290-1 standard method. An interesting observation was that the L. monocytogenes detection by the real-time PCR method was less affected in the presence of Listeria innocua in the contaminated samples, proving therefore to be more reliable than the reference method. The results of this international trial demonstrate that the evaluated real-time PCR-based method represents an excellent alterative to the ISO standard since it shows a higher performance as well as reduce the extent of the analytical process, and can be easily implemented routinely by the competent authorities and food industry laboratories.

U2 - 10.1016/j.ijfoodmicro.2013.12.021

DO - 10.1016/j.ijfoodmicro.2013.12.021

M3 - Journal article

C2 - 24468028

SN - 0168-1605

VL - 184

SP - 128

EP - 133

JO - International Journal of Food Microbiology

JF - International Journal of Food Microbiology

ER -

European validation of a real-time PCR-based method for detection of Listeria monocytogenes in soft cheese

Abstract

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