Interlaboratory validation of a multiplex qPCR method for the detection of Listeria monocytogenes in a ready-to-eat seafood product

Listeria monocytogenes is a major foodborne pathogen which mainly infects susceptible individuals through the consumption of contaminated foods. To this end, ready-to-eat (RTE) food products are of particular concern as this microorganism is widely distributed, can survive, and even grow, under adve...

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Published inFood control Vol. 150; p. 109769
Main Authors Azinheiro, Sarah, Rodríguez-López, Pedro, Lozano-León, Antonio, Guedes, Hugo, Regal, Patricia, Franco, Carlos M., Cepeda, Alberto, Teixeira, Pilar, Melo, Luís D.R., Silva, Daniela, Fernández, Ana, Faria, Márcia, Roumani, Foteini, Herrera, Juan, Prado, Marta, López-Cabo, Marta, Garrido-Maestu, Alejandro
Format Journal Article
LanguageEnglish
Published Elsevier Ltd 01.08.2023
Subjects
Alternative methods
Fish products
Interlaboratory validation
Listeria monocytogenes
qPCR
Ready-to-eat
Fish products
Listeria monocytogenes
qPCR
Ready-to-eat
Interlaboratory validation
Alternative methods
Online AccessGet full text

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Abstract Listeria monocytogenes is a major foodborne pathogen which mainly infects susceptible individuals through the consumption of contaminated foods. To this end, ready-to-eat (RTE) food products are of particular concern as this microorganism is widely distributed, can survive, and even grow, under adverse conditions, and thus must be carefully controlled. In the present study, an interlaboratory ring trial was organized to evaluate an open formula qPCR-based method for the detection of L. monocytogenes. The molecular method was evaluated on a novel RTE seafood product, developed in the framework of a European project, the SEAFOODAGE (EAPA_758/2018). Six laboratories located in Spain and Portugal participated in the study, and the results obtained indicated that this new method presented high diagnostic sensitivity (100%) reaching a low limit of detection (<10 CFU/25 g) with an overall agreement with the reference method, attending to the Cohen's k, of 0.97 that is interpreted as “almost complete agreement”. •An open formula qPCR assay to control L. monocytogenes in a novel ready-to-eat seafood product was evaluated.•The evaluation was performed in an international interlaboratory ring trial.•The assay targets the hly gene and includes a competitive internal amplification control.•It was possible to detect < 10 CFU/25 g by all the participating laboratories.•The Cohen's k obtained was 0.97, interpreted as “almost complete agreement”.
AbstractList Listeria monocytogenes is a major foodborne pathogen which mainly infects susceptible individuals through the consumption of contaminated foods. To this end, ready-to-eat (RTE) food products are of particular concern as this microorganism is widely distributed, can survive, and even grow, under adverse conditions, and thus must be carefully controlled. In the present study, an interlaboratory ring trial was organized to evaluate an open formula qPCR-based method for the detection of L. monocytogenes. The molecular method was evaluated on a novel RTE seafood product, developed in the framework of a European project, the SEAFOODAGE (EAPA_758/2018). Six laboratories located in Spain and Portugal participated in the study, and the results obtained indicated that this new method presented high diagnostic sensitivity (100%) reaching a low limit of detection (<10 CFU/25 g) with an overall agreement with the reference method, attending to the Cohen's k, of 0.97 that is interpreted as “almost complete agreement”. •An open formula qPCR assay to control L. monocytogenes in a novel ready-to-eat seafood product was evaluated.•The evaluation was performed in an international interlaboratory ring trial.•The assay targets the hly gene and includes a competitive internal amplification control.•It was possible to detect < 10 CFU/25 g by all the participating laboratories.•The Cohen's k obtained was 0.97, interpreted as “almost complete agreement”.
ArticleNumber 109769
Author Azinheiro, Sarah
Faria, Márcia
Roumani, Foteini
Rodríguez-López, Pedro
Melo, Luís D.R.
Silva, Daniela
Lozano-León, Antonio
Regal, Patricia
Franco, Carlos M.
Prado, Marta
Cepeda, Alberto
Fernández, Ana
Garrido-Maestu, Alejandro
Teixeira, Pilar
López-Cabo, Marta
Guedes, Hugo
Herrera, Juan
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  surname: Franco
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  surname: Roumani
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  surname: Herrera
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  email: alejandro.garrido@inl.int
  organization: Food Quality & Safety Research Group. International Iberian Nanotechnology Laboratory, Av. Mestre José Veiga s/n, 4715-330, Braga, Portugal
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Keywords Fish products
Listeria monocytogenes
qPCR
Ready-to-eat
Interlaboratory validation
Alternative methods
Language English
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Snippet Listeria monocytogenes is a major foodborne pathogen which mainly infects susceptible individuals through the consumption of contaminated foods. To this end,...
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elsevier
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Publisher
StartPage 109769
SubjectTerms Alternative methods
Fish products
Interlaboratory validation
Listeria monocytogenes
qPCR
Ready-to-eat
Title Interlaboratory validation of a multiplex qPCR method for the detection of Listeria monocytogenes in a ready-to-eat seafood product
URI https://dx.doi.org/10.1016/j.foodcont.2023.109769
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