Quadruplex Real-Time TaqMan<sup>®</sup> 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...
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| Format: | Article |
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MDPI AG
2023-07-01
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| Series: | Viruses |
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| Online Access: | https://www.mdpi.com/1999-4915/15/8/1626 |
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| author | Mariano Carossino Udeni B. R. Balasuriya Côme J. Thieulent Maria E. Barrandeguy Maria Aldana Vissani Viviana Parreño |
| author_facet | Mariano Carossino Udeni B. R. Balasuriya Côme J. Thieulent Maria E. Barrandeguy Maria Aldana Vissani Viviana Parreño |
| author_sort | Mariano Carossino |
| collection | DOAJ |
| 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<sup>®</sup> 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 (<i>p</i>-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. |
| first_indexed | 2024-03-10T23:31:09Z |
| format | Article |
| id | doaj.art-e240ce346da14735b9a8ec17ade9dff4 |
| institution | Directory Open Access Journal |
| issn | 1999-4915 |
| language | English |
| last_indexed | 2024-03-10T23:31:09Z |
| publishDate | 2023-07-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Viruses |
| spelling | doaj.art-e240ce346da14735b9a8ec17ade9dff42023-11-19T03:19:22ZengMDPI AGViruses1999-49152023-07-01158162610.3390/v15081626Quadruplex Real-Time TaqMan<sup>®</sup> RT-qPCR Assay for Differentiation of Equine Group A and B Rotaviruses and Identification of Group A G3 and G14 GenotypesMariano Carossino0Udeni B. R. Balasuriya1Côme J. Thieulent2Maria E. Barrandeguy3Maria Aldana Vissani4Viviana Parreño5Louisiana Animal Disease Diagnostic Laboratory, School of Veterinary Medicine, Louisiana State University, Baton Rouge, LA 70803, USALouisiana Animal Disease Diagnostic Laboratory, School of Veterinary Medicine, Louisiana State University, Baton Rouge, LA 70803, USALouisiana Animal Disease Diagnostic Laboratory, School of Veterinary Medicine, Louisiana State University, Baton Rouge, LA 70803, USAEscuela de Veterinaria, Universidad del Salvador, Buenos Aires B1630, ArgentinaEscuela de Veterinaria, Universidad del Salvador, Buenos Aires B1630, ArgentinaInstituto de Virología, CICVyA, Instituto Nacional de Tecnología Agropecuaria (INTA), Buenos Aires B1686, ArgentinaEquine 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<sup>®</sup> 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 (<i>p</i>-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.https://www.mdpi.com/1999-4915/15/8/1626rotavirus Aequine rotavirus AERVArotavirus Bequine rotavirus BERVB |
| spellingShingle | Mariano Carossino Udeni B. R. Balasuriya Côme J. Thieulent Maria E. Barrandeguy Maria Aldana Vissani Viviana Parreño Quadruplex Real-Time TaqMan<sup>®</sup> RT-qPCR Assay for Differentiation of Equine Group A and B Rotaviruses and Identification of Group A G3 and G14 Genotypes Viruses rotavirus A equine rotavirus A ERVA rotavirus B equine rotavirus B ERVB |
| title | Quadruplex Real-Time TaqMan<sup>®</sup> 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<sup>®</sup> 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<sup>®</sup> 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<sup>®</sup> 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<sup>®</sup> 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 sup r sup rt qpcr assay for differentiation of equine group a and b rotaviruses and identification of group a g3 and g14 genotypes |
| topic | rotavirus A equine rotavirus A ERVA rotavirus B equine rotavirus B ERVB |
| url | https://www.mdpi.com/1999-4915/15/8/1626 |
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