A Field-Applicable Method for Bluetongue Virus Detection Using Filter Paper Cards

Autores
Quiroga, Estefanía; Roldán, Julieta Suyai; Cardoso, Nancy; Sala, Juan Manuel; Marucho, Stefanía Selene; Ferrufino, Cecilia Gabriela; Dus Santos, Maria Jose
Año de publicación
2026
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
The diagnosis of Bluetongue Virus (BTV) in Argentina is primarily conducted via RT-qPCR and faces challenges due to the limited number of diagnostic laboratories, many of which are located in regions where BTV is exotic and far from primary sample collection areas. As a result, blood samples must be transported over considerable distances for analysis. This poses significant logistical challenges due to the specific transport conditions that are required. Consequently, the development of methodologies that optimize sample collection, preservation, and safe transport is of vital importance. The objective of this study was to standardize a sampling protocol for the detection of BTV using filter paper cards. This protocol was developed to ensure adequate preservation and transport of blood samples for diagnosis. Hydration with TE at 37 °C resulted in the lowest Cq among the tested treatments, with ~10 TCID50/mL representing the lowest viral concentration that was consistently detected under the experimental conditions. The stability of the sample on the cards was evaluated through a series of storage trials at temperatures of 25 °C, 4 °C, −20 °C, and −80 °C for 1, 7, 30, 60, 120, and 360 days. Stability was verified up to 120 days at all temperatures. Furthermore, the absence of infectious material in the card eluate was confirmed. A preliminary field evaluation was conducted, in which blood samples from cattle and sheep were collected in tubes and on cards. According to the Cohen’s Kappa index, the agreement between both methods was nearly perfect. The standardized methodology signifies a substantial advancement in BTV diagnosis, as it facilitates the transportation of samples without the necessity of refrigeration and with adequate biosafety conditions without losing diagnostic capacity.
Instituto de Virología
Fil: Quiroga, Estefanía. Universidad Nacional de Hurlingham. Instituto de Biotecnología; Argentina
Fil: Quiroga, Estefanía. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Virología e Innovaciones Tecnológicas; Argentina
Fil: Roldán, Julieta Suyai. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Virología e Innovaciones Tecnológicas; Argentina
Fil: Cardoso, Nancy Patricia. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Virología e Innovaciones Tecnológicas; Argentina
Fil: Cardoso, Nancy Patricia. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Sala, Juan Manuel. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Mercedes; Argentina
Fil: Marucho, Stefanía Selene. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Virología e Innovaciones Tecnológicas; Argentina
Fil: Ferrufino, Cecilia Gabriela. Universidad Nacional de Hurlingham. Instituto de Biotecnología; Argentina
Fil: Ferrufino, Cecilia Gabriela. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Virología e Innovaciones Tecnológicas; Argentina
Fil: Dus Santos, María José. Universidad Nacional de Hurlingham. Instituto de Biotecnología; Argentina
Fil: Dus Santos, María José. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Virología e Innovaciones Tecnológicas; Argentina
Fil: Dus Santos, María José. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fuente
Veterinary Sciences 13 (9) : 955. (September 2026)
Materia
Enfermedades de los Animales
Virus Lengua Azul
Diagnóstico
Filtro
Bioseguridad
Animal Diseases
Bluetongue Virus
Diagnosis
Filters
Biosecurity
Nivel de accesibilidad
acceso abierto
Condiciones de uso
http://creativecommons.org/licenses/by-nc-sa/4.0/
Repositorio
INTA Digital (INTA)
Institución
Instituto Nacional de Tecnología Agropecuaria
OAI Identificador
oai:localhost:20.500.12123/27888
Acceder
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oai_identifier_str oai:localhost:20.500.12123/27888
network_acronym_str INTADig
repository_id_str l
network_name_str INTA Digital (INTA)
spelling A Field-Applicable Method for Bluetongue Virus Detection Using Filter Paper CardsQuiroga, EstefaníaRoldán, Julieta SuyaiCardoso, NancySala, Juan ManuelMarucho, Stefanía SeleneFerrufino, Cecilia GabrielaDus Santos, Maria JoseEnfermedades de los AnimalesVirus Lengua AzulDiagnósticoFiltroBioseguridadAnimal DiseasesBluetongue VirusDiagnosisFiltersBiosecurityThe diagnosis of Bluetongue Virus (BTV) in Argentina is primarily conducted via RT-qPCR and faces challenges due to the limited number of diagnostic laboratories, many of which are located in regions where BTV is exotic and far from primary sample collection areas. As a result, blood samples must be transported over considerable distances for analysis. This poses significant logistical challenges due to the specific transport conditions that are required. Consequently, the development of methodologies that optimize sample collection, preservation, and safe transport is of vital importance. The objective of this study was to standardize a sampling protocol for the detection of BTV using filter paper cards. This protocol was developed to ensure adequate preservation and transport of blood samples for diagnosis. Hydration with TE at 37 °C resulted in the lowest Cq among the tested treatments, with ~10 TCID50/mL representing the lowest viral concentration that was consistently detected under the experimental conditions. The stability of the sample on the cards was evaluated through a series of storage trials at temperatures of 25 °C, 4 °C, −20 °C, and −80 °C for 1, 7, 30, 60, 120, and 360 days. Stability was verified up to 120 days at all temperatures. Furthermore, the absence of infectious material in the card eluate was confirmed. A preliminary field evaluation was conducted, in which blood samples from cattle and sheep were collected in tubes and on cards. According to the Cohen’s Kappa index, the agreement between both methods was nearly perfect. The standardized methodology signifies a substantial advancement in BTV diagnosis, as it facilitates the transportation of samples without the necessity of refrigeration and with adequate biosafety conditions without losing diagnostic capacity.Instituto de VirologíaFil: Quiroga, Estefanía. Universidad Nacional de Hurlingham. Instituto de Biotecnología; ArgentinaFil: Quiroga, Estefanía. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Virología e Innovaciones Tecnológicas; ArgentinaFil: Roldán, Julieta Suyai. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Virología e Innovaciones Tecnológicas; ArgentinaFil: Cardoso, Nancy Patricia. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Virología e Innovaciones Tecnológicas; ArgentinaFil: Cardoso, Nancy Patricia. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Sala, Juan Manuel. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Mercedes; ArgentinaFil: Marucho, Stefanía Selene. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Virología e Innovaciones Tecnológicas; ArgentinaFil: Ferrufino, Cecilia Gabriela. Universidad Nacional de Hurlingham. Instituto de Biotecnología; ArgentinaFil: Ferrufino, Cecilia Gabriela. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Virología e Innovaciones Tecnológicas; ArgentinaFil: Dus Santos, María José. Universidad Nacional de Hurlingham. Instituto de Biotecnología; ArgentinaFil: Dus Santos, María José. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Virología e Innovaciones Tecnológicas; ArgentinaFil: Dus Santos, María José. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaMDPI2026-09-22T13:59:00Z2026-09-22T13:59:00Z2026-09info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfhttp://hdl.handle.net/20.500.12123/27888https://www.mdpi.com/2306-7381/13/9/9552306-7381https://doi.org/10.3390/vetsci13090955Veterinary Sciences 13 (9) : 955. (September 2026)reponame:INTA Digital (INTA)instname:Instituto Nacional de Tecnología Agropecuariaenginfo:eu-repograntAgreement/INTA/2023-PD-L06-I114, Desarrollo y aplicación de métodos diagnósticos y epidemiológicos para la producción pecuaria sustentable y agroalimentaria en humanos con foco en Una Saludinfo:eu-repo/semantics/openAccesshttp://creativecommons.org/licenses/by-nc-sa/4.0/Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International (CC BY-NC-SA 4.0)2026-09-24T11:43:52Zoai:localhost:20.500.12123/27888instacron:INTAInstitucionalhttp://repositorio.inta.gob.ar/Organismo científico-tecnológicoNo correspondehttp://repositorio.inta.gob.ar/oai/requesttripaldi.nicolas@inta.gob.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:l2026-09-24 11:43:52.777INTA Digital (INTA) - Instituto Nacional de Tecnología Agropecuariafalse
dc.title.none.fl_str_mv A Field-Applicable Method for Bluetongue Virus Detection Using Filter Paper Cards
title A Field-Applicable Method for Bluetongue Virus Detection Using Filter Paper Cards
spellingShingle A Field-Applicable Method for Bluetongue Virus Detection Using Filter Paper Cards
Quiroga, Estefanía
Enfermedades de los Animales
Virus Lengua Azul
Diagnóstico
Filtro
Bioseguridad
Animal Diseases
Bluetongue Virus
Diagnosis
Filters
Biosecurity
title_short A Field-Applicable Method for Bluetongue Virus Detection Using Filter Paper Cards
title_full A Field-Applicable Method for Bluetongue Virus Detection Using Filter Paper Cards
title_fullStr A Field-Applicable Method for Bluetongue Virus Detection Using Filter Paper Cards
title_full_unstemmed A Field-Applicable Method for Bluetongue Virus Detection Using Filter Paper Cards
title_sort A Field-Applicable Method for Bluetongue Virus Detection Using Filter Paper Cards
dc.creator.none.fl_str_mv Quiroga, Estefanía
Roldán, Julieta Suyai
Cardoso, Nancy
Sala, Juan Manuel
Marucho, Stefanía Selene
Ferrufino, Cecilia Gabriela
Dus Santos, Maria Jose
author Quiroga, Estefanía
author_facet Quiroga, Estefanía
Roldán, Julieta Suyai
Cardoso, Nancy
Sala, Juan Manuel
Marucho, Stefanía Selene
Ferrufino, Cecilia Gabriela
Dus Santos, Maria Jose
author_role author
author2 Roldán, Julieta Suyai
Cardoso, Nancy
Sala, Juan Manuel
Marucho, Stefanía Selene
Ferrufino, Cecilia Gabriela
Dus Santos, Maria Jose
author2_role author
author
author
author
author
author
dc.subject.none.fl_str_mv Enfermedades de los Animales
Virus Lengua Azul
Diagnóstico
Filtro
Bioseguridad
Animal Diseases
Bluetongue Virus
Diagnosis
Filters
Biosecurity
topic Enfermedades de los Animales
Virus Lengua Azul
Diagnóstico
Filtro
Bioseguridad
Animal Diseases
Bluetongue Virus
Diagnosis
Filters
Biosecurity
dc.description.none.fl_txt_mv The diagnosis of Bluetongue Virus (BTV) in Argentina is primarily conducted via RT-qPCR and faces challenges due to the limited number of diagnostic laboratories, many of which are located in regions where BTV is exotic and far from primary sample collection areas. As a result, blood samples must be transported over considerable distances for analysis. This poses significant logistical challenges due to the specific transport conditions that are required. Consequently, the development of methodologies that optimize sample collection, preservation, and safe transport is of vital importance. The objective of this study was to standardize a sampling protocol for the detection of BTV using filter paper cards. This protocol was developed to ensure adequate preservation and transport of blood samples for diagnosis. Hydration with TE at 37 °C resulted in the lowest Cq among the tested treatments, with ~10 TCID50/mL representing the lowest viral concentration that was consistently detected under the experimental conditions. The stability of the sample on the cards was evaluated through a series of storage trials at temperatures of 25 °C, 4 °C, −20 °C, and −80 °C for 1, 7, 30, 60, 120, and 360 days. Stability was verified up to 120 days at all temperatures. Furthermore, the absence of infectious material in the card eluate was confirmed. A preliminary field evaluation was conducted, in which blood samples from cattle and sheep were collected in tubes and on cards. According to the Cohen’s Kappa index, the agreement between both methods was nearly perfect. The standardized methodology signifies a substantial advancement in BTV diagnosis, as it facilitates the transportation of samples without the necessity of refrigeration and with adequate biosafety conditions without losing diagnostic capacity.
Instituto de Virología
Fil: Quiroga, Estefanía. Universidad Nacional de Hurlingham. Instituto de Biotecnología; Argentina
Fil: Quiroga, Estefanía. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Virología e Innovaciones Tecnológicas; Argentina
Fil: Roldán, Julieta Suyai. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Virología e Innovaciones Tecnológicas; Argentina
Fil: Cardoso, Nancy Patricia. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Virología e Innovaciones Tecnológicas; Argentina
Fil: Cardoso, Nancy Patricia. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Sala, Juan Manuel. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Mercedes; Argentina
Fil: Marucho, Stefanía Selene. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Virología e Innovaciones Tecnológicas; Argentina
Fil: Ferrufino, Cecilia Gabriela. Universidad Nacional de Hurlingham. Instituto de Biotecnología; Argentina
Fil: Ferrufino, Cecilia Gabriela. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Virología e Innovaciones Tecnológicas; Argentina
Fil: Dus Santos, María José. Universidad Nacional de Hurlingham. Instituto de Biotecnología; Argentina
Fil: Dus Santos, María José. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Virología e Innovaciones Tecnológicas; Argentina
Fil: Dus Santos, María José. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
description The diagnosis of Bluetongue Virus (BTV) in Argentina is primarily conducted via RT-qPCR and faces challenges due to the limited number of diagnostic laboratories, many of which are located in regions where BTV is exotic and far from primary sample collection areas. As a result, blood samples must be transported over considerable distances for analysis. This poses significant logistical challenges due to the specific transport conditions that are required. Consequently, the development of methodologies that optimize sample collection, preservation, and safe transport is of vital importance. The objective of this study was to standardize a sampling protocol for the detection of BTV using filter paper cards. This protocol was developed to ensure adequate preservation and transport of blood samples for diagnosis. Hydration with TE at 37 °C resulted in the lowest Cq among the tested treatments, with ~10 TCID50/mL representing the lowest viral concentration that was consistently detected under the experimental conditions. The stability of the sample on the cards was evaluated through a series of storage trials at temperatures of 25 °C, 4 °C, −20 °C, and −80 °C for 1, 7, 30, 60, 120, and 360 days. Stability was verified up to 120 days at all temperatures. Furthermore, the absence of infectious material in the card eluate was confirmed. A preliminary field evaluation was conducted, in which blood samples from cattle and sheep were collected in tubes and on cards. According to the Cohen’s Kappa index, the agreement between both methods was nearly perfect. The standardized methodology signifies a substantial advancement in BTV diagnosis, as it facilitates the transportation of samples without the necessity of refrigeration and with adequate biosafety conditions without losing diagnostic capacity.
publishDate 2026
dc.date.none.fl_str_mv 2026-09-22T13:59:00Z
2026-09-22T13:59:00Z
2026-09
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/20.500.12123/27888
https://www.mdpi.com/2306-7381/13/9/955
2306-7381
https://doi.org/10.3390/vetsci13090955
url http://hdl.handle.net/20.500.12123/27888
https://www.mdpi.com/2306-7381/13/9/955
https://doi.org/10.3390/vetsci13090955
identifier_str_mv 2306-7381
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repograntAgreement/INTA/2023-PD-L06-I114, Desarrollo y aplicación de métodos diagnósticos y epidemiológicos para la producción pecuaria sustentable y agroalimentaria en humanos con foco en Una Salud
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
http://creativecommons.org/licenses/by-nc-sa/4.0/
Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International (CC BY-NC-SA 4.0)
eu_rights_str_mv openAccess
rights_invalid_str_mv http://creativecommons.org/licenses/by-nc-sa/4.0/
Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International (CC BY-NC-SA 4.0)
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv MDPI
publisher.none.fl_str_mv MDPI
dc.source.none.fl_str_mv Veterinary Sciences 13 (9) : 955. (September 2026)
reponame:INTA Digital (INTA)
instname:Instituto Nacional de Tecnología Agropecuaria
reponame_str INTA Digital (INTA)
collection INTA Digital (INTA)
instname_str Instituto Nacional de Tecnología Agropecuaria
repository.name.fl_str_mv INTA Digital (INTA) - Instituto Nacional de Tecnología Agropecuaria
repository.mail.fl_str_mv tripaldi.nicolas@inta.gob.ar
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