Droplet Digital PCR Enhances Sensitivity of Canine Distemper Virus Detection
Canine distemper virus (CDV) poses a substantial threat to diverse carnivorans, leading to systemic and often fatal diseases. Accurate and prompt diagnosis is paramount for effective management and curbing further transmission. This study evaluates the diagnostic performance of droplet digital PCR (...
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| Format: | article |
| Language: | English |
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2024
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| Online Access: | https://hdl.handle.net/20.500.12008/51913 https://doi.org/10.3390/v16111720 |
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| _version_ | 1868890178867691520 |
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| author | Iribarnegaray Perera, María Victoria |
| author2 | Godiño Morales, Guillermo Jesús Larrañaga Sposito, Camila Yamasaki, Kanji Verdes García, José Manuel Puentes Palombo, Rodrigo Eduardo |
| author2_role | author author author author author |
| author_browse | Godiño Morales, Guillermo Jesús Iribarnegaray Perera, María Victoria Larrañaga Sposito, Camila Puentes Palombo, Rodrigo Eduardo Verdes García, José Manuel Yamasaki, Kanji |
| author_facet | Iribarnegaray Perera, María Victoria Godiño Morales, Guillermo Jesús Larrañaga Sposito, Camila Yamasaki, Kanji Verdes García, José Manuel Puentes Palombo, Rodrigo Eduardo |
| author_role | author |
| collection | COLIBRI |
| dc.contributor.none.fl_str_mv | Iribarnegaray Perera María Victoria, Universidad de la República (Uruguay). Veterinary Faculty. Department of Pathobiology. Microbiology Unit Godiño Morales Guillermo Jesús, Universidad de la República (Uruguay). Veterinary Faculty. Department of Pathobiology. Pathology Unit Larrañaga Sposito Camila, Universidad de la República (Uruguay). Veterinary Faculty. Department of Pathobiology. Pathology Unit Yamasaki Kanji, Universidad de la República (Uruguay). Veterinary Faculty. Department of Pathobiology. Pathology Unit Verdes García José Manuel, Universidad de la República (Uruguay). Veterinary Faculty. Department of Pathobiology. Pathology Unit Puentes Palombo Rodrigo Eduardo, Universidad de la República (Uruguay). Veterinary Faculty. Department of Pathobiology. Microbiology Unit |
| dc.creator.none.fl_str_mv | Iribarnegaray Perera, María Victoria Godiño Morales, Guillermo Jesús Larrañaga Sposito, Camila Yamasaki, Kanji Verdes García, José Manuel Puentes Palombo, Rodrigo Eduardo |
| dc.date.none.fl_str_mv | 2024 2025-10-06T12:57:59Z 2025-10-06T12:57:59Z |
| dc.format.none.fl_str_mv | 14 p application/pdf |
| dc.identifier.none.fl_str_mv | Iribarnegaray Perera, M, Godiño Morales, G, Larrañaga Sposito, C, Yamasaki, K, Verdes García, J y Puentes Palombo, R. Droplet Digital PCR Enhances Sensitivity of Canine Distemper Virus Detection. Viruses. [en línea] 2024, 16(11), 1720, 1-14 https://hdl.handle.net/20.500.12008/51913 https://doi.org/10.3390/v16111720 |
| dc.language.none.fl_str_mv | en eng |
| dc.relation.none.fl_str_mv | Viruses, 2024, 16(11), 1720, 1-14 |
| dc.rights.none.fl_str_mv | info:eu-repo/semantics/openAccess Licencia Creative Commons Atribución (CC - By 4.0) |
| dc.source.none.fl_str_mv | reponame:COLIBRI instname:Universidad de la República instacron:Universidad de la República |
| dc.subject.none.fl_str_mv | VIROSIS TECNICAS DE DIAGNOSTICO PCR |
| dc.title.none.fl_str_mv | Droplet Digital PCR Enhances Sensitivity of Canine Distemper Virus Detection |
| dc.type.none.fl_str_mv | Artículo info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion |
| description | Canine distemper virus (CDV) poses a substantial threat to diverse carnivorans, leading to systemic and often fatal diseases. Accurate and prompt diagnosis is paramount for effective management and curbing further transmission. This study evaluates the diagnostic performance of droplet digital PCR (ddPCR) in comparison to conventional reverse-transcription (RT-PCR) and quantitative reverse-transcription real-time PCR (RT-qPCR). Seventy-six clinical samples were collected from dogs with CDV symptoms diagnosed by specialized veterinarians, and sixteen samples from apparently healthy individuals. Conventional PCR, quantitative real-time PCR, and ddPCR were deployed, and their diagnostic capabilities were meticulously assessed. DdPCR exhibited heightened analytical sensitivity, reaching a detection limit of 3 copies/μL, whereas RT-qPCR had a detection limit of 86 copies/μL. The comparative analysis between clinical diagnosis and molecular techniques, including RT-PCR and RT-qPCR, demonstrated low concordance, with Kappa coefficients of 0.268 and 0.324, respectively. In contrast, ddPCR showed a moderate concordance, with a Kappa coefficient of 0.477. The sensitivity was 42.4% for RT-PCR, 57.9% for RT-qPCR, and 72.4% for ddPCR, with 100% specificity for all methods. This study underscores ddPCR’s superior sensitivity and agreement with clinical CDV diagnosis, even at low viral concentrations, suggesting it as a promising alternative for CDV diagnosis. |
| eu_rights_str_mv | openAccess |
| format | article |
| id | anni_bdba3b4da6d85a3e200d38dcc40edf38 |
| identifier_str_mv | Iribarnegaray Perera, M, Godiño Morales, G, Larrañaga Sposito, C, Yamasaki, K, Verdes García, J y Puentes Palombo, R. Droplet Digital PCR Enhances Sensitivity of Canine Distemper Virus Detection. Viruses. [en línea] 2024, 16(11), 1720, 1-14 |
| instacron_str | Universidad de la República |
| institution | Universidad de la República |
| instname_str | Universidad de la República |
| language | eng |
| language_invalid_str_mv | en |
| network_acronym_str | anni |
| network_name_str | oai-lr-anni |
| oai_identifier_str | oai:colibri.udelar.edu.uy:20.500.12008/51913 |
| publishDate | 2024 |
| publishDateSort | 2024 |
| reponame_str | COLIBRI |
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| rights_invalid_str_mv | Licencia Creative Commons Atribución (CC - By 4.0) |
| spelling | Droplet Digital PCR Enhances Sensitivity of Canine Distemper Virus DetectionIribarnegaray Perera, María VictoriaGodiño Morales, Guillermo JesúsLarrañaga Sposito, CamilaYamasaki, KanjiVerdes García, José ManuelPuentes Palombo, Rodrigo EduardoVIROSISTECNICAS DE DIAGNOSTICOPCRCanine distemper virus (CDV) poses a substantial threat to diverse carnivorans, leading to systemic and often fatal diseases. Accurate and prompt diagnosis is paramount for effective management and curbing further transmission. This study evaluates the diagnostic performance of droplet digital PCR (ddPCR) in comparison to conventional reverse-transcription (RT-PCR) and quantitative reverse-transcription real-time PCR (RT-qPCR). Seventy-six clinical samples were collected from dogs with CDV symptoms diagnosed by specialized veterinarians, and sixteen samples from apparently healthy individuals. Conventional PCR, quantitative real-time PCR, and ddPCR were deployed, and their diagnostic capabilities were meticulously assessed. DdPCR exhibited heightened analytical sensitivity, reaching a detection limit of 3 copies/μL, whereas RT-qPCR had a detection limit of 86 copies/μL. The comparative analysis between clinical diagnosis and molecular techniques, including RT-PCR and RT-qPCR, demonstrated low concordance, with Kappa coefficients of 0.268 and 0.324, respectively. In contrast, ddPCR showed a moderate concordance, with a Kappa coefficient of 0.477. The sensitivity was 42.4% for RT-PCR, 57.9% for RT-qPCR, and 72.4% for ddPCR, with 100% specificity for all methods. This study underscores ddPCR’s superior sensitivity and agreement with clinical CDV diagnosis, even at low viral concentrations, suggesting it as a promising alternative for CDV diagnosis.Iribarnegaray Perera María Victoria, Universidad de la República (Uruguay). Veterinary Faculty. Department of Pathobiology. Microbiology UnitGodiño Morales Guillermo Jesús, Universidad de la República (Uruguay). Veterinary Faculty. Department of Pathobiology. Pathology UnitLarrañaga Sposito Camila, Universidad de la República (Uruguay). Veterinary Faculty. Department of Pathobiology. Pathology UnitYamasaki Kanji, Universidad de la República (Uruguay). Veterinary Faculty. Department of Pathobiology. Pathology UnitVerdes García José Manuel, Universidad de la República (Uruguay). Veterinary Faculty. Department of Pathobiology. Pathology UnitPuentes Palombo Rodrigo Eduardo, Universidad de la República (Uruguay). Veterinary Faculty. Department of Pathobiology. Microbiology Unit2025-10-06T12:57:59Z2025-10-06T12:57:59Z2024Artículoinfo:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersion14 papplication/pdfIribarnegaray Perera, M, Godiño Morales, G, Larrañaga Sposito, C, Yamasaki, K, Verdes García, J y Puentes Palombo, R. Droplet Digital PCR Enhances Sensitivity of Canine Distemper Virus Detection. Viruses. [en línea] 2024, 16(11), 1720, 1-14https://hdl.handle.net/20.500.12008/51913https://doi.org/10.3390/v16111720reponame:COLIBRIinstname:Universidad de la Repúblicainstacron:Universidad de la RepúblicaenengViruses, 2024, 16(11), 1720, 1-14Las obras depositadas en el Repositorio se rigen por la Ordenanza de los Derechos de la Propiedad Intelectual de la Universidad de la República.(Res. Nº 91 de C.D.C. de 8/III/1994 – D.O. 7/IV/1994) y por la Ordenanza del Repositorio Abierto de la Universidad de la República (Res. Nº 16 de C.D.C. de 07/10/2014)info:eu-repo/semantics/openAccessLicencia Creative Commons Atribución (CC - By 4.0)oai:colibri.udelar.edu.uy:20.500.12008/519132026-04-14T10:35:09Z |
| spellingShingle | Droplet Digital PCR Enhances Sensitivity of Canine Distemper Virus Detection Iribarnegaray Perera, María Victoria VIROSIS TECNICAS DE DIAGNOSTICO PCR |
| status_str | publishedVersion |
| title | Droplet Digital PCR Enhances Sensitivity of Canine Distemper Virus Detection |
| title_full | Droplet Digital PCR Enhances Sensitivity of Canine Distemper Virus Detection |
| title_fullStr | Droplet Digital PCR Enhances Sensitivity of Canine Distemper Virus Detection |
| title_full_unstemmed | Droplet Digital PCR Enhances Sensitivity of Canine Distemper Virus Detection |
| title_short | Droplet Digital PCR Enhances Sensitivity of Canine Distemper Virus Detection |
| title_sort | Droplet Digital PCR Enhances Sensitivity of Canine Distemper Virus Detection |
| topic | VIROSIS TECNICAS DE DIAGNOSTICO PCR |
| url | https://hdl.handle.net/20.500.12008/51913 https://doi.org/10.3390/v16111720 |