Development of a qPCR to evaluate accurately the treatment response of dogs with demodicosis

Autores
Martínez, Sofía; Sastre, Natalia; Francino, Olga; Maté, María Laura; Sanchez Bruni, Sergio Fabian; del Sole, Maria Jose; Ferrer, Lluis
Año de publicación
2020
Idioma
inglés
Tipo de recurso
documento de conferencia
Estado
versión publicada
Descripción
Canine demodicosis is the result of an overpopulation of Demodex mites, normal inhabitants of most skin of mammals. There is no reliable method to quantify the number of Demodex mites present in the skin to provide accurate information about treatment response. The objective of this study was to develop a molecular technique to quantify the Demodex load in canine skin in order to evaluate the conventional treatment response in dogs with demodicosis. For this purpose, a real time qPCR was developed using primers that amplified a fragment of the Demodex canis 18S rRNA gene. Ten-fold dilutions of the DNA extracted from isolated mites were used to elaborate the standard curve. In addition, 44 skin biopsies (4 mm) obtained from nine canine cadavers and 18 skin biopsies from seven dogs with demodicosis in days zero and 14, 35 and 56 post-treatment (Animal Welfare Committee, FCV-UNCPBA) were used to assess the number of mites in the skin of healthy dogs and of dogs with demodicosis. The skin biopsies of healthy dogs were negative in 38 samples, being positive (>one mite) the remaining samples (six). However, in skin biopsies of diseased dogs the initial Demodex load was mostly >100 mites, decreasing to 50-100, 10-50 and 0-10 mites at 14, 35 and 56 days post-treatment, respectively. The homology between the qPCR product and the 18S rRNA gene was confirmed by DNA sequencing (GenBank KC010485 - Demodex canis). This simple technique could be a useful tool to evaluate accurately the response to treatment of dogs with demodicosis.
Fil: Martínez, Sofía. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tandil. Centro de Investigación Veterinaria de Tandil. Universidad Nacional del Centro de la Provincia de Buenos Aires. Centro de Investigación Veterinaria de Tandil. Provincia de Buenos Aires. Gobernación. Comision de Investigaciones Científicas. Centro de Investigación Veterinaria de Tandil; Argentina
Fil: Sastre, Natalia. Universidad Autonoma de Barcelona. Facultad de Veterinaria; España
Fil: Francino, Olga. Universidad Autonoma de Barcelona. Facultad de Veterinaria; España
Fil: Maté, María Laura. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tandil. Centro de Investigación Veterinaria de Tandil. Universidad Nacional del Centro de la Provincia de Buenos Aires. Centro de Investigación Veterinaria de Tandil. Provincia de Buenos Aires. Gobernación. Comision de Investigaciones Científicas. Centro de Investigación Veterinaria de Tandil; Argentina
Fil: Sanchez Bruni, Sergio Fabian. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tandil. Centro de Investigación Veterinaria de Tandil. Universidad Nacional del Centro de la Provincia de Buenos Aires. Centro de Investigación Veterinaria de Tandil. Provincia de Buenos Aires. Gobernación. Comision de Investigaciones Científicas. Centro de Investigación Veterinaria de Tandil; Argentina
Fil: del Sole, Maria Jose. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tandil. Centro de Investigación Veterinaria de Tandil. Universidad Nacional del Centro de la Provincia de Buenos Aires. Centro de Investigación Veterinaria de Tandil. Provincia de Buenos Aires. Gobernación. Comision de Investigaciones Científicas. Centro de Investigación Veterinaria de Tandil; Argentina
Fil: Ferrer, Lluis. Universidad Autonoma de Barcelona. Facultad de Veterinaria; España
9th World Congress Of Veterinary Dermatology
Australia
World Association of Veterinary Dermatology
Materia
DEMODICOSIS
Q-PCR
MAMMALS
IVERMECTIN
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:11336/154007
Acceder
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network_name_str CONICET Digital (CONICET)
spelling Development of a qPCR to evaluate accurately the treatment response of dogs with demodicosisMartínez, SofíaSastre, NataliaFrancino, OlgaMaté, María LauraSanchez Bruni, Sergio Fabiandel Sole, Maria JoseFerrer, LluisDEMODICOSISQ-PCRMAMMALSIVERMECTINhttps://purl.org/becyt/ford/4.3https://purl.org/becyt/ford/4Canine demodicosis is the result of an overpopulation of Demodex mites, normal inhabitants of most skin of mammals. There is no reliable method to quantify the number of Demodex mites present in the skin to provide accurate information about treatment response. The objective of this study was to develop a molecular technique to quantify the Demodex load in canine skin in order to evaluate the conventional treatment response in dogs with demodicosis. For this purpose, a real time qPCR was developed using primers that amplified a fragment of the Demodex canis 18S rRNA gene. Ten-fold dilutions of the DNA extracted from isolated mites were used to elaborate the standard curve. In addition, 44 skin biopsies (4 mm) obtained from nine canine cadavers and 18 skin biopsies from seven dogs with demodicosis in days zero and 14, 35 and 56 post-treatment (Animal Welfare Committee, FCV-UNCPBA) were used to assess the number of mites in the skin of healthy dogs and of dogs with demodicosis. The skin biopsies of healthy dogs were negative in 38 samples, being positive (>one mite) the remaining samples (six). However, in skin biopsies of diseased dogs the initial Demodex load was mostly >100 mites, decreasing to 50-100, 10-50 and 0-10 mites at 14, 35 and 56 days post-treatment, respectively. The homology between the qPCR product and the 18S rRNA gene was confirmed by DNA sequencing (GenBank KC010485 - Demodex canis). This simple technique could be a useful tool to evaluate accurately the response to treatment of dogs with demodicosis.Fil: Martínez, Sofía. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tandil. Centro de Investigación Veterinaria de Tandil. Universidad Nacional del Centro de la Provincia de Buenos Aires. Centro de Investigación Veterinaria de Tandil. Provincia de Buenos Aires. Gobernación. Comision de Investigaciones Científicas. Centro de Investigación Veterinaria de Tandil; ArgentinaFil: Sastre, Natalia. Universidad Autonoma de Barcelona. Facultad de Veterinaria; EspañaFil: Francino, Olga. Universidad Autonoma de Barcelona. Facultad de Veterinaria; EspañaFil: Maté, María Laura. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tandil. Centro de Investigación Veterinaria de Tandil. Universidad Nacional del Centro de la Provincia de Buenos Aires. Centro de Investigación Veterinaria de Tandil. Provincia de Buenos Aires. Gobernación. Comision de Investigaciones Científicas. Centro de Investigación Veterinaria de Tandil; ArgentinaFil: Sanchez Bruni, Sergio Fabian. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tandil. Centro de Investigación Veterinaria de Tandil. Universidad Nacional del Centro de la Provincia de Buenos Aires. Centro de Investigación Veterinaria de Tandil. Provincia de Buenos Aires. Gobernación. Comision de Investigaciones Científicas. Centro de Investigación Veterinaria de Tandil; ArgentinaFil: del Sole, Maria Jose. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tandil. Centro de Investigación Veterinaria de Tandil. Universidad Nacional del Centro de la Provincia de Buenos Aires. Centro de Investigación Veterinaria de Tandil. Provincia de Buenos Aires. Gobernación. Comision de Investigaciones Científicas. Centro de Investigación Veterinaria de Tandil; ArgentinaFil: Ferrer, Lluis. Universidad Autonoma de Barcelona. Facultad de Veterinaria; España9th World Congress Of Veterinary DermatologyAustraliaWorld Association of Veterinary DermatologyWiley Blackwell Publishing, Inc2020info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/conferenceObjectCongresoJournalhttp://purl.org/coar/resource_type/c_5794info:ar-repo/semantics/documentoDeConferenciaapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/154007Development of a qPCR to evaluate accurately the treatment response of dogs with demodicosis; 9th World Congress Of Veterinary Dermatology; Australia; 2020; 26-261365-3164CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/https://onlinelibrary.wiley.com/doi/epdf/10.1111/vde.12907info:eu-repo/semantics/altIdentifier/doi/10.1111/vde.12907Internacionalinfo:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-29T09:50:47Zoai:ri.conicet.gov.ar:11336/154007instacron: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 09:50:48.218CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Development of a qPCR to evaluate accurately the treatment response of dogs with demodicosis
title Development of a qPCR to evaluate accurately the treatment response of dogs with demodicosis
spellingShingle Development of a qPCR to evaluate accurately the treatment response of dogs with demodicosis
Martínez, Sofía
DEMODICOSIS
Q-PCR
MAMMALS
IVERMECTIN
title_short Development of a qPCR to evaluate accurately the treatment response of dogs with demodicosis
title_full Development of a qPCR to evaluate accurately the treatment response of dogs with demodicosis
title_fullStr Development of a qPCR to evaluate accurately the treatment response of dogs with demodicosis
title_full_unstemmed Development of a qPCR to evaluate accurately the treatment response of dogs with demodicosis
title_sort Development of a qPCR to evaluate accurately the treatment response of dogs with demodicosis
dc.creator.none.fl_str_mv Martínez, Sofía
Sastre, Natalia
Francino, Olga
Maté, María Laura
Sanchez Bruni, Sergio Fabian
del Sole, Maria Jose
Ferrer, Lluis
author Martínez, Sofía
author_facet Martínez, Sofía
Sastre, Natalia
Francino, Olga
Maté, María Laura
Sanchez Bruni, Sergio Fabian
del Sole, Maria Jose
Ferrer, Lluis
author_role author
author2 Sastre, Natalia
Francino, Olga
Maté, María Laura
Sanchez Bruni, Sergio Fabian
del Sole, Maria Jose
Ferrer, Lluis
author2_role author
author
author
author
author
author
dc.subject.none.fl_str_mv DEMODICOSIS
Q-PCR
MAMMALS
IVERMECTIN
topic DEMODICOSIS
Q-PCR
MAMMALS
IVERMECTIN
purl_subject.fl_str_mv https://purl.org/becyt/ford/4.3
https://purl.org/becyt/ford/4
dc.description.none.fl_txt_mv Canine demodicosis is the result of an overpopulation of Demodex mites, normal inhabitants of most skin of mammals. There is no reliable method to quantify the number of Demodex mites present in the skin to provide accurate information about treatment response. The objective of this study was to develop a molecular technique to quantify the Demodex load in canine skin in order to evaluate the conventional treatment response in dogs with demodicosis. For this purpose, a real time qPCR was developed using primers that amplified a fragment of the Demodex canis 18S rRNA gene. Ten-fold dilutions of the DNA extracted from isolated mites were used to elaborate the standard curve. In addition, 44 skin biopsies (4 mm) obtained from nine canine cadavers and 18 skin biopsies from seven dogs with demodicosis in days zero and 14, 35 and 56 post-treatment (Animal Welfare Committee, FCV-UNCPBA) were used to assess the number of mites in the skin of healthy dogs and of dogs with demodicosis. The skin biopsies of healthy dogs were negative in 38 samples, being positive (>one mite) the remaining samples (six). However, in skin biopsies of diseased dogs the initial Demodex load was mostly >100 mites, decreasing to 50-100, 10-50 and 0-10 mites at 14, 35 and 56 days post-treatment, respectively. The homology between the qPCR product and the 18S rRNA gene was confirmed by DNA sequencing (GenBank KC010485 - Demodex canis). This simple technique could be a useful tool to evaluate accurately the response to treatment of dogs with demodicosis.
Fil: Martínez, Sofía. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tandil. Centro de Investigación Veterinaria de Tandil. Universidad Nacional del Centro de la Provincia de Buenos Aires. Centro de Investigación Veterinaria de Tandil. Provincia de Buenos Aires. Gobernación. Comision de Investigaciones Científicas. Centro de Investigación Veterinaria de Tandil; Argentina
Fil: Sastre, Natalia. Universidad Autonoma de Barcelona. Facultad de Veterinaria; España
Fil: Francino, Olga. Universidad Autonoma de Barcelona. Facultad de Veterinaria; España
Fil: Maté, María Laura. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tandil. Centro de Investigación Veterinaria de Tandil. Universidad Nacional del Centro de la Provincia de Buenos Aires. Centro de Investigación Veterinaria de Tandil. Provincia de Buenos Aires. Gobernación. Comision de Investigaciones Científicas. Centro de Investigación Veterinaria de Tandil; Argentina
Fil: Sanchez Bruni, Sergio Fabian. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tandil. Centro de Investigación Veterinaria de Tandil. Universidad Nacional del Centro de la Provincia de Buenos Aires. Centro de Investigación Veterinaria de Tandil. Provincia de Buenos Aires. Gobernación. Comision de Investigaciones Científicas. Centro de Investigación Veterinaria de Tandil; Argentina
Fil: del Sole, Maria Jose. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tandil. Centro de Investigación Veterinaria de Tandil. Universidad Nacional del Centro de la Provincia de Buenos Aires. Centro de Investigación Veterinaria de Tandil. Provincia de Buenos Aires. Gobernación. Comision de Investigaciones Científicas. Centro de Investigación Veterinaria de Tandil; Argentina
Fil: Ferrer, Lluis. Universidad Autonoma de Barcelona. Facultad de Veterinaria; España
9th World Congress Of Veterinary Dermatology
Australia
World Association of Veterinary Dermatology
description Canine demodicosis is the result of an overpopulation of Demodex mites, normal inhabitants of most skin of mammals. There is no reliable method to quantify the number of Demodex mites present in the skin to provide accurate information about treatment response. The objective of this study was to develop a molecular technique to quantify the Demodex load in canine skin in order to evaluate the conventional treatment response in dogs with demodicosis. For this purpose, a real time qPCR was developed using primers that amplified a fragment of the Demodex canis 18S rRNA gene. Ten-fold dilutions of the DNA extracted from isolated mites were used to elaborate the standard curve. In addition, 44 skin biopsies (4 mm) obtained from nine canine cadavers and 18 skin biopsies from seven dogs with demodicosis in days zero and 14, 35 and 56 post-treatment (Animal Welfare Committee, FCV-UNCPBA) were used to assess the number of mites in the skin of healthy dogs and of dogs with demodicosis. The skin biopsies of healthy dogs were negative in 38 samples, being positive (>one mite) the remaining samples (six). However, in skin biopsies of diseased dogs the initial Demodex load was mostly >100 mites, decreasing to 50-100, 10-50 and 0-10 mites at 14, 35 and 56 days post-treatment, respectively. The homology between the qPCR product and the 18S rRNA gene was confirmed by DNA sequencing (GenBank KC010485 - Demodex canis). This simple technique could be a useful tool to evaluate accurately the response to treatment of dogs with demodicosis.
publishDate 2020
dc.date.none.fl_str_mv 2020
dc.type.none.fl_str_mv info:eu-repo/semantics/publishedVersion
info:eu-repo/semantics/conferenceObject
Congreso
Journal
http://purl.org/coar/resource_type/c_5794
info:ar-repo/semantics/documentoDeConferencia
status_str publishedVersion
format conferenceObject
dc.identifier.none.fl_str_mv http://hdl.handle.net/11336/154007
Development of a qPCR to evaluate accurately the treatment response of dogs with demodicosis; 9th World Congress Of Veterinary Dermatology; Australia; 2020; 26-26
1365-3164
CONICET Digital
CONICET
url http://hdl.handle.net/11336/154007
identifier_str_mv Development of a qPCR to evaluate accurately the treatment response of dogs with demodicosis; 9th World Congress Of Veterinary Dermatology; Australia; 2020; 26-26
1365-3164
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/epdf/10.1111/vde.12907
info:eu-repo/semantics/altIdentifier/doi/10.1111/vde.12907
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
dc.coverage.none.fl_str_mv Internacional
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
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