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
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 diarr...
| Autores principales: | , , , , , |
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| Formato: | Artículo |
| Lenguaje: | Inglés |
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MDPI
2023
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| Materias: | |
| Acceso en línea: | http://hdl.handle.net/20.500.12123/15205 https://www.mdpi.com/1999-4915/15/8/1626 https://doi.org/10.3390/v15081626 |
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| author | Carossino, Mariano Balasuriya, Udeni B.R. Thieulent, Côme J. Barrandeguy, Maria Edith Vissani, Maria Aldana Parreño, Gladys Viviana |
| author_browse | Balasuriya, Udeni B.R. Barrandeguy, Maria Edith Carossino, Mariano Parreño, Gladys Viviana Thieulent, Côme J. Vissani, Maria Aldana |
| author_facet | Carossino, Mariano Balasuriya, Udeni B.R. Thieulent, Côme J. Barrandeguy, Maria Edith Vissani, Maria Aldana Parreño, Gladys Viviana |
| author_sort | Carossino, Mariano |
| collection | INTA Digital |
| 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. |
| format | Artículo |
| id | INTA15205 |
| institution | Instituto Nacional de Tecnología Agropecuaria (INTA -Argentina) |
| language | Inglés |
| publishDate | 2023 |
| publishDateRange | 2023 |
| publishDateSort | 2023 |
| publisher | MDPI |
| publisherStr | MDPI |
| record_format | dspace |
| spelling | INTA152052023-09-14T10:46:28Z 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 Balasuriya, Udeni B.R. Thieulent, Côme J. Barrandeguy, Maria Edith Vissani, Maria Aldana Parreño, Gladys Viviana Rotavirus Diarrhoea PCR Equidae Genotypes Diarrea Genotipos Equine Rotavirus A Equine Rotavirus B Rotavirus A Equino Rotavirus B Equino 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 2023-09-14T10:35:36Z 2023-09-14T10:35:36Z 2023-07 info:ar-repo/semantics/artículo info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion 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 eng 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) application/pdf MDPI Viruses 15 (8) : 1626 (Agosto 2023) |
| spellingShingle | Rotavirus Diarrhoea PCR Equidae Genotypes Diarrea Genotipos Equine Rotavirus A Equine Rotavirus B Rotavirus A Equino Rotavirus B Equino Carossino, Mariano Balasuriya, Udeni B.R. Thieulent, Côme J. Barrandeguy, Maria Edith Vissani, Maria Aldana Parreño, Gladys Viviana 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 |
| 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_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_sort | quadruplex real time taqman r rt qpcr assay for differentiation of equine group a and b rotaviruses and identification of group a g3 and g14 genotypes |
| topic | Rotavirus Diarrhoea PCR Equidae Genotypes Diarrea Genotipos Equine Rotavirus A Equine Rotavirus B Rotavirus A Equino Rotavirus B Equino |
| url | http://hdl.handle.net/20.500.12123/15205 https://www.mdpi.com/1999-4915/15/8/1626 https://doi.org/10.3390/v15081626 |
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