Optical and electrochemical detection of a verotoxigenic E. coli gene using DNAzyme-labeled stem-loops

Autores
Longinotti, Gloria; Ybarra, Gabriel Omar; Montserrat, Javier Marcelo
Año de publicación
2017
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
The activity of a peroxidase-mimicking DNAzyme was optimized to be used as a catalytic label in a stem-loop genosensor construction for quantifying the gene sequence Shiga-like toxin I of verotoxigenic E. coli. Experimental conditions such as pH, buffer composition, potassium ion concentration, and hemin-to-oligonucleotides ratio, were analyzed to maximize optical and electrochemical responses using microvolumes. Different stem-loop constructions were evaluated to obtain the optimum response against the target concentration. Linear ranges of 0.05-0.5 µM and limits of detection of 174 nM and 144 nM were estimated for the optical and electrochemical measurements, respectively. Selectivity was proved by assaying other verotoxigenic, enterotoxigenic and enteroinvasive sequences. The results show that, if a combination of small-volume electrochemical cells and low-cost untreated screen-printed electrodes with a relatively high geometric area is used, electrochemical measurements present similar sensitivity and limits of detection to the more usual optical ones, allowing the development of low-cost electrochemical biosensors based on the use of soluble DNAzymes as labels.
Fil: Longinotti, Gloria. Instituto Nacional de Tecnología Industrial; Argentina
Fil: Ybarra, Gabriel Omar. Instituto Nacional de Tecnología Industrial; Argentina
Fil: Montserrat, Javier Marcelo. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de General Sarmiento; Argentina
Materia
biosensor
genosensor
DNAzyme peroxidase
vertoxigenic E. coli
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:11336/74499
Acceder
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oai_identifier_str oai:ri.conicet.gov.ar:11336/74499
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Optical and electrochemical detection of a verotoxigenic E. coli gene using DNAzyme-labeled stem-loopsLonginotti, GloriaYbarra, Gabriel OmarMontserrat, Javier MarcelobiosensorgenosensorDNAzyme peroxidasevertoxigenic E. colihttps://purl.org/becyt/ford/1.4https://purl.org/becyt/ford/1The activity of a peroxidase-mimicking DNAzyme was optimized to be used as a catalytic label in a stem-loop genosensor construction for quantifying the gene sequence Shiga-like toxin I of verotoxigenic E. coli. Experimental conditions such as pH, buffer composition, potassium ion concentration, and hemin-to-oligonucleotides ratio, were analyzed to maximize optical and electrochemical responses using microvolumes. Different stem-loop constructions were evaluated to obtain the optimum response against the target concentration. Linear ranges of 0.05-0.5 µM and limits of detection of 174 nM and 144 nM were estimated for the optical and electrochemical measurements, respectively. Selectivity was proved by assaying other verotoxigenic, enterotoxigenic and enteroinvasive sequences. The results show that, if a combination of small-volume electrochemical cells and low-cost untreated screen-printed electrodes with a relatively high geometric area is used, electrochemical measurements present similar sensitivity and limits of detection to the more usual optical ones, allowing the development of low-cost electrochemical biosensors based on the use of soluble DNAzymes as labels.Fil: Longinotti, Gloria. Instituto Nacional de Tecnología Industrial; ArgentinaFil: Ybarra, Gabriel Omar. Instituto Nacional de Tecnología Industrial; ArgentinaFil: Montserrat, Javier Marcelo. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de General Sarmiento; ArgentinaInternational Association of Physical Chemists2017-12info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/vnd.openxmlformats-officedocument.wordprocessingml.documentapplication/pdfhttp://hdl.handle.net/11336/74499Longinotti, Gloria; Ybarra, Gabriel Omar; Montserrat, Javier Marcelo; Optical and electrochemical detection of a verotoxigenic E. coli gene using DNAzyme-labeled stem-loops; International Association of Physical Chemists; Journal of Electrochemical Science and Engineering; 7; 4; 12-2017; 193-2001847-9286CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/doi/10.5599/jese.471info:eu-repo/semantics/altIdentifier/url/http://pub.iapchem.org/ojs/index.php/JESE/article/view/471info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-03T10:02:03Zoai:ri.conicet.gov.ar:11336/74499instacron: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:34982025-09-03 10:02:03.568CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Optical and electrochemical detection of a verotoxigenic E. coli gene using DNAzyme-labeled stem-loops
title Optical and electrochemical detection of a verotoxigenic E. coli gene using DNAzyme-labeled stem-loops
spellingShingle Optical and electrochemical detection of a verotoxigenic E. coli gene using DNAzyme-labeled stem-loops
Longinotti, Gloria
biosensor
genosensor
DNAzyme peroxidase
vertoxigenic E. coli
title_short Optical and electrochemical detection of a verotoxigenic E. coli gene using DNAzyme-labeled stem-loops
title_full Optical and electrochemical detection of a verotoxigenic E. coli gene using DNAzyme-labeled stem-loops
title_fullStr Optical and electrochemical detection of a verotoxigenic E. coli gene using DNAzyme-labeled stem-loops
title_full_unstemmed Optical and electrochemical detection of a verotoxigenic E. coli gene using DNAzyme-labeled stem-loops
title_sort Optical and electrochemical detection of a verotoxigenic E. coli gene using DNAzyme-labeled stem-loops
dc.creator.none.fl_str_mv Longinotti, Gloria
Ybarra, Gabriel Omar
Montserrat, Javier Marcelo
author Longinotti, Gloria
author_facet Longinotti, Gloria
Ybarra, Gabriel Omar
Montserrat, Javier Marcelo
author_role author
author2 Ybarra, Gabriel Omar
Montserrat, Javier Marcelo
author2_role author
author
dc.subject.none.fl_str_mv biosensor
genosensor
DNAzyme peroxidase
vertoxigenic E. coli
topic biosensor
genosensor
DNAzyme peroxidase
vertoxigenic E. coli
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.4
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv The activity of a peroxidase-mimicking DNAzyme was optimized to be used as a catalytic label in a stem-loop genosensor construction for quantifying the gene sequence Shiga-like toxin I of verotoxigenic E. coli. Experimental conditions such as pH, buffer composition, potassium ion concentration, and hemin-to-oligonucleotides ratio, were analyzed to maximize optical and electrochemical responses using microvolumes. Different stem-loop constructions were evaluated to obtain the optimum response against the target concentration. Linear ranges of 0.05-0.5 µM and limits of detection of 174 nM and 144 nM were estimated for the optical and electrochemical measurements, respectively. Selectivity was proved by assaying other verotoxigenic, enterotoxigenic and enteroinvasive sequences. The results show that, if a combination of small-volume electrochemical cells and low-cost untreated screen-printed electrodes with a relatively high geometric area is used, electrochemical measurements present similar sensitivity and limits of detection to the more usual optical ones, allowing the development of low-cost electrochemical biosensors based on the use of soluble DNAzymes as labels.
Fil: Longinotti, Gloria. Instituto Nacional de Tecnología Industrial; Argentina
Fil: Ybarra, Gabriel Omar. Instituto Nacional de Tecnología Industrial; Argentina
Fil: Montserrat, Javier Marcelo. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de General Sarmiento; Argentina
description The activity of a peroxidase-mimicking DNAzyme was optimized to be used as a catalytic label in a stem-loop genosensor construction for quantifying the gene sequence Shiga-like toxin I of verotoxigenic E. coli. Experimental conditions such as pH, buffer composition, potassium ion concentration, and hemin-to-oligonucleotides ratio, were analyzed to maximize optical and electrochemical responses using microvolumes. Different stem-loop constructions were evaluated to obtain the optimum response against the target concentration. Linear ranges of 0.05-0.5 µM and limits of detection of 174 nM and 144 nM were estimated for the optical and electrochemical measurements, respectively. Selectivity was proved by assaying other verotoxigenic, enterotoxigenic and enteroinvasive sequences. The results show that, if a combination of small-volume electrochemical cells and low-cost untreated screen-printed electrodes with a relatively high geometric area is used, electrochemical measurements present similar sensitivity and limits of detection to the more usual optical ones, allowing the development of low-cost electrochemical biosensors based on the use of soluble DNAzymes as labels.
publishDate 2017
dc.date.none.fl_str_mv 2017-12
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/11336/74499
Longinotti, Gloria; Ybarra, Gabriel Omar; Montserrat, Javier Marcelo; Optical and electrochemical detection of a verotoxigenic E. coli gene using DNAzyme-labeled stem-loops; International Association of Physical Chemists; Journal of Electrochemical Science and Engineering; 7; 4; 12-2017; 193-200
1847-9286
CONICET Digital
CONICET
url http://hdl.handle.net/11336/74499
identifier_str_mv Longinotti, Gloria; Ybarra, Gabriel Omar; Montserrat, Javier Marcelo; Optical and electrochemical detection of a verotoxigenic E. coli gene using DNAzyme-labeled stem-loops; International Association of Physical Chemists; Journal of Electrochemical Science and Engineering; 7; 4; 12-2017; 193-200
1847-9286
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/doi/10.5599/jese.471
info:eu-repo/semantics/altIdentifier/url/http://pub.iapchem.org/ojs/index.php/JESE/article/view/471
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/vnd.openxmlformats-officedocument.wordprocessingml.document
application/pdf
dc.publisher.none.fl_str_mv International Association of Physical Chemists
publisher.none.fl_str_mv International Association of Physical Chemists
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_ 1842269734253363200
score 13.13397

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