Quadruplex real-time TaqMan® RT-qPCR assay for differentiation of equine group A and B rotaviruses and Identification of group A G3 and G14 genotypes

Autores
Carossino, Mariano; Balasuriya, Udeni B.R.; Thieulent, Côme J.; Barrandeguy, Maria Edith; Vissani, Maria Aldana; Parreño, Gladys Viviana
Año de publicación
2023
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Equine rotavirus A (ERVA) is the leading cause of diarrhea in foals, with G3P[12] and G14P[12] genotypes being the most prevalent. Recently, equine G3-like RVA was recognized as an emerging infection in children, and a group B equine rotavirus (ERVB) was identified as an emergent cause of foal diarrhea in the US. Thus, there is a need to adapt molecular diagnostic tools for improved detection and surveillance to identify emerging strains, understand their molecular epidemiology, and inform future vaccine development. We developed a quadruplex TaqMan® RT-qPCR assay for differentiation of ERVA and ERVB and simultaneous G-typing of ERVA strains, evaluated its analytical and clinical performance, and compared it to (1) a previously established ERVA triplex RT-qPCR assay and (2) standard RT-PCR assay and Sanger sequencing of PCR products. This quadruplex RT-qPCR assay demonstrated high sensitivity (>90%)/specificity (100%) for every target and high overall agreement (>96%). Comparison between the triplex and quadruplex assays revealed only a slightly higher sensitivity for the ERVA NSP3 target using the triplex format (p-value 0.008) while no significant differences were detected for other targets. This quadruplex RT-qPCR assay will significantly enhance rapid surveillance of both ERVA and ERVB circulating and emerging strains with potential for interspecies transmission.
Instituto de Virología
Fil: Carossino, Mariano. Louisiana State University. School of Veterinary Medicine. Louisiana Animal Disease Diagnostic Laboratory and Department of Pathobiological Sciences; Estados Unidos
Fil: Balasuriya, Udeni B. R. Louisiana State University. School of Veterinary Medicine. Louisiana Animal Disease Diagnostic Laboratory and Department of Pathobiological Sciences; Estados Unidos
Fil: Thieulent, Côme J. Louisiana State University. School of Veterinary Medicine. Louisiana Animal Disease Diagnostic Laboratory and Department of Pathobiological Sciences; Estados Unidos
Fil: Barrandeguy, Maria Edith. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Virología; Argentina
Fil: Barrandeguy, Maria Edith. Universidad del Salvador. Escuela de Veterinaria; Argentina
Fil: Vissani, Aldana. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Virología: Argentina
Fil: Vissani, Aldana. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Vissani, Aldana. Universidad del Salvador. Escuela de Veterinaria; Argentina
Fil: Parreño, Gladys Viviana. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Virología; Argentina
Fil: Parreño, Gladys Viviana. Consejo Nacional de Investigaciones Cientificas y Tecnicas; Argentina
Fuente
Viruses 15 (8) : 1626 (Agosto 2023)
Materia
Rotavirus
Diarrhoea
PCR
Equidae
Genotypes
Diarrea
Genotipos
Equine Rotavirus A
Equine Rotavirus B
Rotavirus A Equino
Rotavirus B Equino
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
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spelling Quadruplex real-time TaqMan® RT-qPCR assay for differentiation of equine group A and B rotaviruses and Identification of group A G3 and G14 genotypesCarossino, MarianoBalasuriya, Udeni B.R.Thieulent, Côme J.Barrandeguy, Maria EdithVissani, Maria AldanaParreño, Gladys VivianaRotavirusDiarrhoeaPCREquidaeGenotypesDiarreaGenotiposEquine Rotavirus AEquine Rotavirus BRotavirus A EquinoRotavirus B EquinoEquine rotavirus A (ERVA) is the leading cause of diarrhea in foals, with G3P[12] and G14P[12] genotypes being the most prevalent. Recently, equine G3-like RVA was recognized as an emerging infection in children, and a group B equine rotavirus (ERVB) was identified as an emergent cause of foal diarrhea in the US. Thus, there is a need to adapt molecular diagnostic tools for improved detection and surveillance to identify emerging strains, understand their molecular epidemiology, and inform future vaccine development. We developed a quadruplex TaqMan® RT-qPCR assay for differentiation of ERVA and ERVB and simultaneous G-typing of ERVA strains, evaluated its analytical and clinical performance, and compared it to (1) a previously established ERVA triplex RT-qPCR assay and (2) standard RT-PCR assay and Sanger sequencing of PCR products. This quadruplex RT-qPCR assay demonstrated high sensitivity (>90%)/specificity (100%) for every target and high overall agreement (>96%). Comparison between the triplex and quadruplex assays revealed only a slightly higher sensitivity for the ERVA NSP3 target using the triplex format (p-value 0.008) while no significant differences were detected for other targets. This quadruplex RT-qPCR assay will significantly enhance rapid surveillance of both ERVA and ERVB circulating and emerging strains with potential for interspecies transmission.Instituto de VirologíaFil: Carossino, Mariano. Louisiana State University. School of Veterinary Medicine. Louisiana Animal Disease Diagnostic Laboratory and Department of Pathobiological Sciences; Estados UnidosFil: Balasuriya, Udeni B. R. Louisiana State University. School of Veterinary Medicine. Louisiana Animal Disease Diagnostic Laboratory and Department of Pathobiological Sciences; Estados UnidosFil: Thieulent, Côme J. Louisiana State University. School of Veterinary Medicine. Louisiana Animal Disease Diagnostic Laboratory and Department of Pathobiological Sciences; Estados UnidosFil: Barrandeguy, Maria Edith. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Virología; ArgentinaFil: Barrandeguy, Maria Edith. Universidad del Salvador. Escuela de Veterinaria; ArgentinaFil: Vissani, Aldana. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Virología: ArgentinaFil: Vissani, Aldana. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Vissani, Aldana. Universidad del Salvador. Escuela de Veterinaria; ArgentinaFil: Parreño, Gladys Viviana. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Virología; ArgentinaFil: Parreño, Gladys Viviana. Consejo Nacional de Investigaciones Cientificas y Tecnicas; ArgentinaMDPI2023-09-14T10:35:36Z2023-09-14T10:35:36Z2023-07info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfhttp://hdl.handle.net/20.500.12123/15205https://www.mdpi.com/1999-4915/15/8/16261999-4915https://doi.org/10.3390/v15081626Viruses 15 (8) : 1626 (Agosto 2023)reponame:INTA Digital (INTA)instname:Instituto Nacional de Tecnología Agropecuariaenginfo: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)2025-09-04T09:49:56Zoai:localhost:20.500.12123/15205instacron:INTAInstitucionalhttp://repositorio.inta.gob.ar/Organismo científico-tecnológicoNo correspondehttp://repositorio.inta.gob.ar/oai/requesttripaldi.nicolas@inta.gob.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:l2025-09-04 09:49:56.644INTA Digital (INTA) - Instituto Nacional de Tecnología Agropecuariafalse
dc.title.none.fl_str_mv Quadruplex real-time TaqMan® RT-qPCR assay for differentiation of equine group A and B rotaviruses and Identification of group A G3 and G14 genotypes
title Quadruplex real-time TaqMan® RT-qPCR assay for differentiation of equine group A and B rotaviruses and Identification of group A G3 and G14 genotypes
spellingShingle Quadruplex real-time TaqMan® RT-qPCR assay for differentiation of equine group A and B rotaviruses and Identification of group A G3 and G14 genotypes
Carossino, Mariano
Rotavirus
Diarrhoea
PCR
Equidae
Genotypes
Diarrea
Genotipos
Equine Rotavirus A
Equine Rotavirus B
Rotavirus A Equino
Rotavirus B Equino
title_short Quadruplex real-time TaqMan® RT-qPCR assay for differentiation of equine group A and B rotaviruses and Identification of group A G3 and G14 genotypes
title_full Quadruplex real-time TaqMan® RT-qPCR assay for differentiation of equine group A and B rotaviruses and Identification of group A G3 and G14 genotypes
title_fullStr Quadruplex real-time TaqMan® RT-qPCR assay for differentiation of equine group A and B rotaviruses and Identification of group A G3 and G14 genotypes
title_full_unstemmed Quadruplex real-time TaqMan® RT-qPCR assay for differentiation of equine group A and B rotaviruses and Identification of group A G3 and G14 genotypes
title_sort Quadruplex real-time TaqMan® RT-qPCR assay for differentiation of equine group A and B rotaviruses and Identification of group A G3 and G14 genotypes
dc.creator.none.fl_str_mv Carossino, Mariano
Balasuriya, Udeni B.R.
Thieulent, Côme J.
Barrandeguy, Maria Edith
Vissani, Maria Aldana
Parreño, Gladys Viviana
author Carossino, Mariano
author_facet Carossino, Mariano
Balasuriya, Udeni B.R.
Thieulent, Côme J.
Barrandeguy, Maria Edith
Vissani, Maria Aldana
Parreño, Gladys Viviana
author_role author
author2 Balasuriya, Udeni B.R.
Thieulent, Côme J.
Barrandeguy, Maria Edith
Vissani, Maria Aldana
Parreño, Gladys Viviana
author2_role author
author
author
author
author
dc.subject.none.fl_str_mv Rotavirus
Diarrhoea
PCR
Equidae
Genotypes
Diarrea
Genotipos
Equine Rotavirus A
Equine Rotavirus B
Rotavirus A Equino
Rotavirus B Equino
topic Rotavirus
Diarrhoea
PCR
Equidae
Genotypes
Diarrea
Genotipos
Equine Rotavirus A
Equine Rotavirus B
Rotavirus A Equino
Rotavirus B Equino
dc.description.none.fl_txt_mv Equine rotavirus A (ERVA) is the leading cause of diarrhea in foals, with G3P[12] and G14P[12] genotypes being the most prevalent. Recently, equine G3-like RVA was recognized as an emerging infection in children, and a group B equine rotavirus (ERVB) was identified as an emergent cause of foal diarrhea in the US. Thus, there is a need to adapt molecular diagnostic tools for improved detection and surveillance to identify emerging strains, understand their molecular epidemiology, and inform future vaccine development. We developed a quadruplex TaqMan® RT-qPCR assay for differentiation of ERVA and ERVB and simultaneous G-typing of ERVA strains, evaluated its analytical and clinical performance, and compared it to (1) a previously established ERVA triplex RT-qPCR assay and (2) standard RT-PCR assay and Sanger sequencing of PCR products. This quadruplex RT-qPCR assay demonstrated high sensitivity (>90%)/specificity (100%) for every target and high overall agreement (>96%). Comparison between the triplex and quadruplex assays revealed only a slightly higher sensitivity for the ERVA NSP3 target using the triplex format (p-value 0.008) while no significant differences were detected for other targets. This quadruplex RT-qPCR assay will significantly enhance rapid surveillance of both ERVA and ERVB circulating and emerging strains with potential for interspecies transmission.
Instituto de Virología
Fil: Carossino, Mariano. Louisiana State University. School of Veterinary Medicine. Louisiana Animal Disease Diagnostic Laboratory and Department of Pathobiological Sciences; Estados Unidos
Fil: Balasuriya, Udeni B. R. Louisiana State University. School of Veterinary Medicine. Louisiana Animal Disease Diagnostic Laboratory and Department of Pathobiological Sciences; Estados Unidos
Fil: Thieulent, Côme J. Louisiana State University. School of Veterinary Medicine. Louisiana Animal Disease Diagnostic Laboratory and Department of Pathobiological Sciences; Estados Unidos
Fil: Barrandeguy, Maria Edith. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Virología; Argentina
Fil: Barrandeguy, Maria Edith. Universidad del Salvador. Escuela de Veterinaria; Argentina
Fil: Vissani, Aldana. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Virología: Argentina
Fil: Vissani, Aldana. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Vissani, Aldana. Universidad del Salvador. Escuela de Veterinaria; Argentina
Fil: Parreño, Gladys Viviana. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Virología; Argentina
Fil: Parreño, Gladys Viviana. Consejo Nacional de Investigaciones Cientificas y Tecnicas; Argentina
description Equine rotavirus A (ERVA) is the leading cause of diarrhea in foals, with G3P[12] and G14P[12] genotypes being the most prevalent. Recently, equine G3-like RVA was recognized as an emerging infection in children, and a group B equine rotavirus (ERVB) was identified as an emergent cause of foal diarrhea in the US. Thus, there is a need to adapt molecular diagnostic tools for improved detection and surveillance to identify emerging strains, understand their molecular epidemiology, and inform future vaccine development. We developed a quadruplex TaqMan® RT-qPCR assay for differentiation of ERVA and ERVB and simultaneous G-typing of ERVA strains, evaluated its analytical and clinical performance, and compared it to (1) a previously established ERVA triplex RT-qPCR assay and (2) standard RT-PCR assay and Sanger sequencing of PCR products. This quadruplex RT-qPCR assay demonstrated high sensitivity (>90%)/specificity (100%) for every target and high overall agreement (>96%). Comparison between the triplex and quadruplex assays revealed only a slightly higher sensitivity for the ERVA NSP3 target using the triplex format (p-value 0.008) while no significant differences were detected for other targets. This quadruplex RT-qPCR assay will significantly enhance rapid surveillance of both ERVA and ERVB circulating and emerging strains with potential for interspecies transmission.
publishDate 2023
dc.date.none.fl_str_mv 2023-09-14T10:35:36Z
2023-09-14T10:35:36Z
2023-07
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/20.500.12123/15205
https://www.mdpi.com/1999-4915/15/8/1626
1999-4915
https://doi.org/10.3390/v15081626
url http://hdl.handle.net/20.500.12123/15205
https://www.mdpi.com/1999-4915/15/8/1626
https://doi.org/10.3390/v15081626
identifier_str_mv 1999-4915
dc.language.none.fl_str_mv eng
language eng
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 Viruses 15 (8) : 1626 (Agosto 2023)
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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