Evaluation of PMA-qPCR methodology to detect and quantify viable Shiga toxin-producing Escherichia coli in beef burgers

Autores
Rey, María de Los Ángeles; Cap, Marina; Favre, Leonardo Cristian; Rodríguez Racca, Anabel; Dus Santos, María José; Vaudagna, Sergio Ramon; Mozgovoj, Marina Valeria
Año de publicación
2021
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Propidium monoazide (PMA) is a selective nucleic acid intercalating dye that can be combined with real-time PCR (qPCR) in order to evaluate cell viability in food samples. The aim of the present work was to evaluate PMA-qPCR to detect and quantify viable STEC cells in beef burgers using stx2 as target gene. First, it was determined that 100 µM of PMA could inhibit qPCR signal from non-viable cells and had no influence on the amplification of different concentrations of viable cells. Then, it was shown that PMA efficiently distinguished between different log cfu of viable cells in presence of a high concentration of non-viable cells, both in culture and in beef burger homogenates. Finally, it was determined that PMA could distinguish between viable and non-viable cells within the same log cfu in beef burger homogenates. PMA-qPCR effectively detected and quantified viable STEC cells in culture and in beef burger homogenates. Novelty impact statement: The main achievement of this work is that we demonstrate PMA-qPCR could not only detect, but also quantify viable STEC cells targeting stx2 gene, even in the presence of a high concentration of non-viable STEC cells in a complex matrix as beef burgers. This methodology can be used to assess effectiveness of antimicrobial treatments to reduce STEC contamination in meat products more rapidly and with less pathogenic residues than conventional methods.
Fil: Rey, María de Los Ángeles. Instituto Nacional de Tecnologia Agropecuaria. Centro de Investigacion de Agroindustria. Instituto de Tecnologia de Alimentos. Instituto de Ciencia y Tecnologia de Sistemas Alimentarios Sustentables. - Consejo Nacional de Investigaciones Cientificas y Tecnicas. Oficina de Coordinacion Administrativa Pque. Centenario. Instituto de Ciencia y Tecnologia de Sistemas Alimentarios Sustentables.; Argentina
Fil: Cap, Marina. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación de Agroindustria. Instituto de Tecnología de Alimentos; Argentina
Fil: Favre, Leonardo Cristian. Instituto Nacional de Tecnologia Agropecuaria. Centro de Investigacion de Agroindustria. Instituto de Tecnologia de Alimentos. Instituto de Ciencia y Tecnologia de Sistemas Alimentarios Sustentables. - Consejo Nacional de Investigaciones Cientificas y Tecnicas. Oficina de Coordinacion Administrativa Pque. Centenario. Instituto de Ciencia y Tecnologia de Sistemas Alimentarios Sustentables.; Argentina
Fil: Rodríguez Racca, Anabel. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario; Argentina
Fil: Dus Santos, María José. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación En Ciencias Veterinarias y Agronómicas. Instituto de Virología E Innovaciones Tecnológicas. - Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Pque. Centenario. Instituto de Virología E Innovaciones Tecnológicas; Argentina
Fil: Vaudagna, Sergio Ramon. Instituto Nacional de Tecnologia Agropecuaria. Centro de Investigacion de Agroindustria. Instituto de Tecnologia de Alimentos. Instituto de Ciencia y Tecnologia de Sistemas Alimentarios Sustentables. - Consejo Nacional de Investigaciones Cientificas y Tecnicas. Oficina de Coordinacion Administrativa Pque. Centenario. Instituto de Ciencia y Tecnologia de Sistemas Alimentarios Sustentables.; Argentina
Fil: Mozgovoj, Marina Valeria. Instituto Nacional de Tecnologia Agropecuaria. Centro de Investigacion de Agroindustria. Instituto de Tecnologia de Alimentos. Instituto de Ciencia y Tecnologia de Sistemas Alimentarios Sustentables. - Consejo Nacional de Investigaciones Cientificas y Tecnicas. Oficina de Coordinacion Administrativa Pque. Centenario. Instituto de Ciencia y Tecnologia de Sistemas Alimentarios Sustentables.; Argentina
Materia
STEC
Propidium monoazide
Viability
qPCR
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/148679
Acceder
id CONICETDig_2f9097ee446cc2c2c8840d90674535dc
oai_identifier_str oai:ri.conicet.gov.ar:11336/148679
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Evaluation of PMA-qPCR methodology to detect and quantify viable Shiga toxin-producing Escherichia coli in beef burgersRey, María de Los ÁngelesCap, MarinaFavre, Leonardo CristianRodríguez Racca, AnabelDus Santos, María JoséVaudagna, Sergio RamonMozgovoj, Marina ValeriaSTECPropidium monoazideViabilityqPCRhttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1https://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1Propidium monoazide (PMA) is a selective nucleic acid intercalating dye that can be combined with real-time PCR (qPCR) in order to evaluate cell viability in food samples. The aim of the present work was to evaluate PMA-qPCR to detect and quantify viable STEC cells in beef burgers using stx2 as target gene. First, it was determined that 100 µM of PMA could inhibit qPCR signal from non-viable cells and had no influence on the amplification of different concentrations of viable cells. Then, it was shown that PMA efficiently distinguished between different log cfu of viable cells in presence of a high concentration of non-viable cells, both in culture and in beef burger homogenates. Finally, it was determined that PMA could distinguish between viable and non-viable cells within the same log cfu in beef burger homogenates. PMA-qPCR effectively detected and quantified viable STEC cells in culture and in beef burger homogenates. Novelty impact statement: The main achievement of this work is that we demonstrate PMA-qPCR could not only detect, but also quantify viable STEC cells targeting stx2 gene, even in the presence of a high concentration of non-viable STEC cells in a complex matrix as beef burgers. This methodology can be used to assess effectiveness of antimicrobial treatments to reduce STEC contamination in meat products more rapidly and with less pathogenic residues than conventional methods.Fil: Rey, María de Los Ángeles. Instituto Nacional de Tecnologia Agropecuaria. Centro de Investigacion de Agroindustria. Instituto de Tecnologia de Alimentos. Instituto de Ciencia y Tecnologia de Sistemas Alimentarios Sustentables. - Consejo Nacional de Investigaciones Cientificas y Tecnicas. Oficina de Coordinacion Administrativa Pque. Centenario. Instituto de Ciencia y Tecnologia de Sistemas Alimentarios Sustentables.; ArgentinaFil: Cap, Marina. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación de Agroindustria. Instituto de Tecnología de Alimentos; ArgentinaFil: Favre, Leonardo Cristian. Instituto Nacional de Tecnologia Agropecuaria. Centro de Investigacion de Agroindustria. Instituto de Tecnologia de Alimentos. Instituto de Ciencia y Tecnologia de Sistemas Alimentarios Sustentables. - Consejo Nacional de Investigaciones Cientificas y Tecnicas. Oficina de Coordinacion Administrativa Pque. Centenario. Instituto de Ciencia y Tecnologia de Sistemas Alimentarios Sustentables.; ArgentinaFil: Rodríguez Racca, Anabel. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario; ArgentinaFil: Dus Santos, María José. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación En Ciencias Veterinarias y Agronómicas. Instituto de Virología E Innovaciones Tecnológicas. - Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Pque. Centenario. Instituto de Virología E Innovaciones Tecnológicas; ArgentinaFil: Vaudagna, Sergio Ramon. Instituto Nacional de Tecnologia Agropecuaria. Centro de Investigacion de Agroindustria. Instituto de Tecnologia de Alimentos. Instituto de Ciencia y Tecnologia de Sistemas Alimentarios Sustentables. - Consejo Nacional de Investigaciones Cientificas y Tecnicas. Oficina de Coordinacion Administrativa Pque. Centenario. Instituto de Ciencia y Tecnologia de Sistemas Alimentarios Sustentables.; ArgentinaFil: Mozgovoj, Marina Valeria. Instituto Nacional de Tecnologia Agropecuaria. Centro de Investigacion de Agroindustria. Instituto de Tecnologia de Alimentos. Instituto de Ciencia y Tecnologia de Sistemas Alimentarios Sustentables. - Consejo Nacional de Investigaciones Cientificas y Tecnicas. Oficina de Coordinacion Administrativa Pque. Centenario. Instituto de Ciencia y Tecnologia de Sistemas Alimentarios Sustentables.; ArgentinaWiley Blackwell Publishing, Inc2021-02info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/148679Rey, María de Los Ángeles; Cap, Marina; Favre, Leonardo Cristian; Rodríguez Racca, Anabel; Dus Santos, María José; et al.; Evaluation of PMA-qPCR methodology to detect and quantify viable Shiga toxin-producing Escherichia coli in beef burgers; Wiley Blackwell Publishing, Inc; Journal of Food Processing and Preservation; 45; 4; 2-2021; 1-80145-8892CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/https://onlinelibrary.wiley.com/doi/10.1111/jfpp.15338info:eu-repo/semantics/altIdentifier/doi/10.1111/jfpp.15338info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-29T10:00:00Zoai:ri.conicet.gov.ar:11336/148679instacron:CONICETInstitucionalhttp://ri.conicet.gov.ar/Organismo científico-tecnológicoNo correspondehttp://ri.conicet.gov.ar/oai/requestdasensio@conicet.gov.ar; lcarlino@conicet.gov.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:34982025-09-29 10:00:00.554CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Evaluation of PMA-qPCR methodology to detect and quantify viable Shiga toxin-producing Escherichia coli in beef burgers
title Evaluation of PMA-qPCR methodology to detect and quantify viable Shiga toxin-producing Escherichia coli in beef burgers
spellingShingle Evaluation of PMA-qPCR methodology to detect and quantify viable Shiga toxin-producing Escherichia coli in beef burgers
Rey, María de Los Ángeles
STEC
Propidium monoazide
Viability
qPCR
title_short Evaluation of PMA-qPCR methodology to detect and quantify viable Shiga toxin-producing Escherichia coli in beef burgers
title_full Evaluation of PMA-qPCR methodology to detect and quantify viable Shiga toxin-producing Escherichia coli in beef burgers
title_fullStr Evaluation of PMA-qPCR methodology to detect and quantify viable Shiga toxin-producing Escherichia coli in beef burgers
title_full_unstemmed Evaluation of PMA-qPCR methodology to detect and quantify viable Shiga toxin-producing Escherichia coli in beef burgers
title_sort Evaluation of PMA-qPCR methodology to detect and quantify viable Shiga toxin-producing Escherichia coli in beef burgers
dc.creator.none.fl_str_mv Rey, María de Los Ángeles
Cap, Marina
Favre, Leonardo Cristian
Rodríguez Racca, Anabel
Dus Santos, María José
Vaudagna, Sergio Ramon
Mozgovoj, Marina Valeria
author Rey, María de Los Ángeles
author_facet Rey, María de Los Ángeles
Cap, Marina
Favre, Leonardo Cristian
Rodríguez Racca, Anabel
Dus Santos, María José
Vaudagna, Sergio Ramon
Mozgovoj, Marina Valeria
author_role author
author2 Cap, Marina
Favre, Leonardo Cristian
Rodríguez Racca, Anabel
Dus Santos, María José
Vaudagna, Sergio Ramon
Mozgovoj, Marina Valeria
author2_role author
author
author
author
author
author
dc.subject.none.fl_str_mv STEC
Propidium monoazide
Viability
qPCR
topic STEC
Propidium monoazide
Viability
qPCR
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv Propidium monoazide (PMA) is a selective nucleic acid intercalating dye that can be combined with real-time PCR (qPCR) in order to evaluate cell viability in food samples. The aim of the present work was to evaluate PMA-qPCR to detect and quantify viable STEC cells in beef burgers using stx2 as target gene. First, it was determined that 100 µM of PMA could inhibit qPCR signal from non-viable cells and had no influence on the amplification of different concentrations of viable cells. Then, it was shown that PMA efficiently distinguished between different log cfu of viable cells in presence of a high concentration of non-viable cells, both in culture and in beef burger homogenates. Finally, it was determined that PMA could distinguish between viable and non-viable cells within the same log cfu in beef burger homogenates. PMA-qPCR effectively detected and quantified viable STEC cells in culture and in beef burger homogenates. Novelty impact statement: The main achievement of this work is that we demonstrate PMA-qPCR could not only detect, but also quantify viable STEC cells targeting stx2 gene, even in the presence of a high concentration of non-viable STEC cells in a complex matrix as beef burgers. This methodology can be used to assess effectiveness of antimicrobial treatments to reduce STEC contamination in meat products more rapidly and with less pathogenic residues than conventional methods.
Fil: Rey, María de Los Ángeles. Instituto Nacional de Tecnologia Agropecuaria. Centro de Investigacion de Agroindustria. Instituto de Tecnologia de Alimentos. Instituto de Ciencia y Tecnologia de Sistemas Alimentarios Sustentables. - Consejo Nacional de Investigaciones Cientificas y Tecnicas. Oficina de Coordinacion Administrativa Pque. Centenario. Instituto de Ciencia y Tecnologia de Sistemas Alimentarios Sustentables.; Argentina
Fil: Cap, Marina. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación de Agroindustria. Instituto de Tecnología de Alimentos; Argentina
Fil: Favre, Leonardo Cristian. Instituto Nacional de Tecnologia Agropecuaria. Centro de Investigacion de Agroindustria. Instituto de Tecnologia de Alimentos. Instituto de Ciencia y Tecnologia de Sistemas Alimentarios Sustentables. - Consejo Nacional de Investigaciones Cientificas y Tecnicas. Oficina de Coordinacion Administrativa Pque. Centenario. Instituto de Ciencia y Tecnologia de Sistemas Alimentarios Sustentables.; Argentina
Fil: Rodríguez Racca, Anabel. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario; Argentina
Fil: Dus Santos, María José. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación En Ciencias Veterinarias y Agronómicas. Instituto de Virología E Innovaciones Tecnológicas. - Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Pque. Centenario. Instituto de Virología E Innovaciones Tecnológicas; Argentina
Fil: Vaudagna, Sergio Ramon. Instituto Nacional de Tecnologia Agropecuaria. Centro de Investigacion de Agroindustria. Instituto de Tecnologia de Alimentos. Instituto de Ciencia y Tecnologia de Sistemas Alimentarios Sustentables. - Consejo Nacional de Investigaciones Cientificas y Tecnicas. Oficina de Coordinacion Administrativa Pque. Centenario. Instituto de Ciencia y Tecnologia de Sistemas Alimentarios Sustentables.; Argentina
Fil: Mozgovoj, Marina Valeria. Instituto Nacional de Tecnologia Agropecuaria. Centro de Investigacion de Agroindustria. Instituto de Tecnologia de Alimentos. Instituto de Ciencia y Tecnologia de Sistemas Alimentarios Sustentables. - Consejo Nacional de Investigaciones Cientificas y Tecnicas. Oficina de Coordinacion Administrativa Pque. Centenario. Instituto de Ciencia y Tecnologia de Sistemas Alimentarios Sustentables.; Argentina
description Propidium monoazide (PMA) is a selective nucleic acid intercalating dye that can be combined with real-time PCR (qPCR) in order to evaluate cell viability in food samples. The aim of the present work was to evaluate PMA-qPCR to detect and quantify viable STEC cells in beef burgers using stx2 as target gene. First, it was determined that 100 µM of PMA could inhibit qPCR signal from non-viable cells and had no influence on the amplification of different concentrations of viable cells. Then, it was shown that PMA efficiently distinguished between different log cfu of viable cells in presence of a high concentration of non-viable cells, both in culture and in beef burger homogenates. Finally, it was determined that PMA could distinguish between viable and non-viable cells within the same log cfu in beef burger homogenates. PMA-qPCR effectively detected and quantified viable STEC cells in culture and in beef burger homogenates. Novelty impact statement: The main achievement of this work is that we demonstrate PMA-qPCR could not only detect, but also quantify viable STEC cells targeting stx2 gene, even in the presence of a high concentration of non-viable STEC cells in a complex matrix as beef burgers. This methodology can be used to assess effectiveness of antimicrobial treatments to reduce STEC contamination in meat products more rapidly and with less pathogenic residues than conventional methods.
publishDate 2021
dc.date.none.fl_str_mv 2021-02
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
http://purl.org/coar/resource_type/c_6501
info:ar-repo/semantics/articulo
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/11336/148679
Rey, María de Los Ángeles; Cap, Marina; Favre, Leonardo Cristian; Rodríguez Racca, Anabel; Dus Santos, María José; et al.; Evaluation of PMA-qPCR methodology to detect and quantify viable Shiga toxin-producing Escherichia coli in beef burgers; Wiley Blackwell Publishing, Inc; Journal of Food Processing and Preservation; 45; 4; 2-2021; 1-8
0145-8892
CONICET Digital
CONICET
url http://hdl.handle.net/11336/148679
identifier_str_mv Rey, María de Los Ángeles; Cap, Marina; Favre, Leonardo Cristian; Rodríguez Racca, Anabel; Dus Santos, María José; et al.; Evaluation of PMA-qPCR methodology to detect and quantify viable Shiga toxin-producing Escherichia coli in beef burgers; Wiley Blackwell Publishing, Inc; Journal of Food Processing and Preservation; 45; 4; 2-2021; 1-8
0145-8892
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/url/https://onlinelibrary.wiley.com/doi/10.1111/jfpp.15338
info:eu-repo/semantics/altIdentifier/doi/10.1111/jfpp.15338
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
eu_rights_str_mv openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
dc.format.none.fl_str_mv application/pdf
application/pdf
application/pdf
application/pdf
dc.publisher.none.fl_str_mv Wiley Blackwell Publishing, Inc
publisher.none.fl_str_mv Wiley Blackwell Publishing, Inc
dc.source.none.fl_str_mv reponame:CONICET Digital (CONICET)
instname:Consejo Nacional de Investigaciones Científicas y Técnicas
reponame_str CONICET Digital (CONICET)
collection CONICET Digital (CONICET)
instname_str Consejo Nacional de Investigaciones Científicas y Técnicas
repository.name.fl_str_mv CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicas
repository.mail.fl_str_mv dasensio@conicet.gov.ar; lcarlino@conicet.gov.ar
_version_ 1844613776418013184
score 13.070432

Publicaciones similares