A Duplex Quantitative Real-Time Reverse Transcription-PCR for Simultaneous Detection and Differentiation of Flaviviruses of the Japanese Encephalitis and Ntaya Serocomplexes in Birds
High impact, mosquito-borne flaviviruses such as West Nile virus (WNV), Usutu virus (USUV), Japanese encephalitis virus (JEV), Tembusu virus (TMUV), and Bagaza/Israel turkey meningoencephalomyelitis virus (BAGV/ITV) are emerging in different areas of the world. These viruses belong to the Japanese e...
| Main Authors: | , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Frontiers Media S.A.
2020-04-01
|
| Series: | Frontiers in Veterinary Science |
| Subjects: | |
| Online Access: | https://www.frontiersin.org/article/10.3389/fvets.2020.00203/full |
| _version_ | 1819106887700840448 |
|---|---|
| author | Maia Elizalde Cristina Cano-Gómez Francisco Llorente Elisa Pérez-Ramírez Laia Casades-Martí Pilar Aguilera-Sepúlveda Francisco Ruiz-Fons Miguel Ángel Jiménez-Clavero Miguel Ángel Jiménez-Clavero Jovita Fernández-Pinero |
| author_facet | Maia Elizalde Cristina Cano-Gómez Francisco Llorente Elisa Pérez-Ramírez Laia Casades-Martí Pilar Aguilera-Sepúlveda Francisco Ruiz-Fons Miguel Ángel Jiménez-Clavero Miguel Ángel Jiménez-Clavero Jovita Fernández-Pinero |
| author_sort | Maia Elizalde |
| collection | DOAJ |
| description | High impact, mosquito-borne flaviviruses such as West Nile virus (WNV), Usutu virus (USUV), Japanese encephalitis virus (JEV), Tembusu virus (TMUV), and Bagaza/Israel turkey meningoencephalomyelitis virus (BAGV/ITV) are emerging in different areas of the world. These viruses belong to the Japanese encephalitis (JE) serocomplex (JEV, WNV, and USUV) and the Ntaya serocomplex (TMUV and BAGV/ITV). Notably, they share transmission route (mosquito bite) and reservoir host type (wild birds), and some of them co-circulate in the same areas, infecting overlapping mosquito and avian population. This may simplify epidemiological surveillance, since it allows the detection of different infections targeting the same population, but also represents a challenge, as the diagnostic tools applied need to detect the whole range of flaviviruses surveyed, and correctly differentiate between these closely related pathogens. To this aim, a duplex real-time RT-PCR (dRRT-PCR) method has been developed for the simultaneous and differential detection of JE and Ntaya flavivirus serocomplexes. The method has been standardized and evaluated by analyzing a panel of 49 flaviviral and non-flaviviral isolates, and clinical samples of different bird species obtained from experimental infections or from the field, proving its value for virus detection in apparently healthy or suspicious animals. This new dRRT-PCR technique is a reliable, specific and highly sensitive tool for rapid detection and differentiation of JE and Ntaya flavivirus groups in either domestic or wild animals. This novel method can be implemented in animal virology diagnostic laboratories as screening tool in routine surveillance and in the event of bird encephalitis emergence. |
| first_indexed | 2024-12-22T02:45:17Z |
| format | Article |
| id | doaj.art-1b269ecd2cf24d0f8cc3cf4655ff5151 |
| institution | Directory Open Access Journal |
| issn | 2297-1769 |
| language | English |
| last_indexed | 2024-12-22T02:45:17Z |
| publishDate | 2020-04-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Veterinary Science |
| spelling | doaj.art-1b269ecd2cf24d0f8cc3cf4655ff51512022-12-21T18:41:32ZengFrontiers Media S.A.Frontiers in Veterinary Science2297-17692020-04-01710.3389/fvets.2020.00203527129A Duplex Quantitative Real-Time Reverse Transcription-PCR for Simultaneous Detection and Differentiation of Flaviviruses of the Japanese Encephalitis and Ntaya Serocomplexes in BirdsMaia Elizalde0Cristina Cano-Gómez1Francisco Llorente2Elisa Pérez-Ramírez3Laia Casades-Martí4Pilar Aguilera-Sepúlveda5Francisco Ruiz-Fons6Miguel Ángel Jiménez-Clavero7Miguel Ángel Jiménez-Clavero8Jovita Fernández-Pinero9Centro de Investigación en Sanidad Animal (CISA), Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria (INIA), Valdeolmos-Alalpardo, SpainCentro de Investigación en Sanidad Animal (CISA), Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria (INIA), Valdeolmos-Alalpardo, SpainCentro de Investigación en Sanidad Animal (CISA), Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria (INIA), Valdeolmos-Alalpardo, SpainCentro de Investigación en Sanidad Animal (CISA), Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria (INIA), Valdeolmos-Alalpardo, SpainInstituto de Investigación de Recursos Cinegéticos IREC (CSIC-UCLM-JCCM), SaBio Group, Ciudad Real, SpainCentro de Investigación en Sanidad Animal (CISA), Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria (INIA), Valdeolmos-Alalpardo, SpainInstituto de Investigación de Recursos Cinegéticos IREC (CSIC-UCLM-JCCM), SaBio Group, Ciudad Real, SpainCentro de Investigación en Sanidad Animal (CISA), Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria (INIA), Valdeolmos-Alalpardo, SpainEpidemiología y Salud Pública, CIBERESP, Madrid, SpainCentro de Investigación en Sanidad Animal (CISA), Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria (INIA), Valdeolmos-Alalpardo, SpainHigh impact, mosquito-borne flaviviruses such as West Nile virus (WNV), Usutu virus (USUV), Japanese encephalitis virus (JEV), Tembusu virus (TMUV), and Bagaza/Israel turkey meningoencephalomyelitis virus (BAGV/ITV) are emerging in different areas of the world. These viruses belong to the Japanese encephalitis (JE) serocomplex (JEV, WNV, and USUV) and the Ntaya serocomplex (TMUV and BAGV/ITV). Notably, they share transmission route (mosquito bite) and reservoir host type (wild birds), and some of them co-circulate in the same areas, infecting overlapping mosquito and avian population. This may simplify epidemiological surveillance, since it allows the detection of different infections targeting the same population, but also represents a challenge, as the diagnostic tools applied need to detect the whole range of flaviviruses surveyed, and correctly differentiate between these closely related pathogens. To this aim, a duplex real-time RT-PCR (dRRT-PCR) method has been developed for the simultaneous and differential detection of JE and Ntaya flavivirus serocomplexes. The method has been standardized and evaluated by analyzing a panel of 49 flaviviral and non-flaviviral isolates, and clinical samples of different bird species obtained from experimental infections or from the field, proving its value for virus detection in apparently healthy or suspicious animals. This new dRRT-PCR technique is a reliable, specific and highly sensitive tool for rapid detection and differentiation of JE and Ntaya flavivirus groups in either domestic or wild animals. This novel method can be implemented in animal virology diagnostic laboratories as screening tool in routine surveillance and in the event of bird encephalitis emergence.https://www.frontiersin.org/article/10.3389/fvets.2020.00203/fullflavivirusJapanese encephalitis serocomplexNtaya serocomplexWest Nile virusdiagnosticsduplex real-time RT-PCR |
| spellingShingle | Maia Elizalde Cristina Cano-Gómez Francisco Llorente Elisa Pérez-Ramírez Laia Casades-Martí Pilar Aguilera-Sepúlveda Francisco Ruiz-Fons Miguel Ángel Jiménez-Clavero Miguel Ángel Jiménez-Clavero Jovita Fernández-Pinero A Duplex Quantitative Real-Time Reverse Transcription-PCR for Simultaneous Detection and Differentiation of Flaviviruses of the Japanese Encephalitis and Ntaya Serocomplexes in Birds Frontiers in Veterinary Science flavivirus Japanese encephalitis serocomplex Ntaya serocomplex West Nile virus diagnostics duplex real-time RT-PCR |
| title | A Duplex Quantitative Real-Time Reverse Transcription-PCR for Simultaneous Detection and Differentiation of Flaviviruses of the Japanese Encephalitis and Ntaya Serocomplexes in Birds |
| title_full | A Duplex Quantitative Real-Time Reverse Transcription-PCR for Simultaneous Detection and Differentiation of Flaviviruses of the Japanese Encephalitis and Ntaya Serocomplexes in Birds |
| title_fullStr | A Duplex Quantitative Real-Time Reverse Transcription-PCR for Simultaneous Detection and Differentiation of Flaviviruses of the Japanese Encephalitis and Ntaya Serocomplexes in Birds |
| title_full_unstemmed | A Duplex Quantitative Real-Time Reverse Transcription-PCR for Simultaneous Detection and Differentiation of Flaviviruses of the Japanese Encephalitis and Ntaya Serocomplexes in Birds |
| title_short | A Duplex Quantitative Real-Time Reverse Transcription-PCR for Simultaneous Detection and Differentiation of Flaviviruses of the Japanese Encephalitis and Ntaya Serocomplexes in Birds |
| title_sort | duplex quantitative real time reverse transcription pcr for simultaneous detection and differentiation of flaviviruses of the japanese encephalitis and ntaya serocomplexes in birds |
| topic | flavivirus Japanese encephalitis serocomplex Ntaya serocomplex West Nile virus diagnostics duplex real-time RT-PCR |
| url | https://www.frontiersin.org/article/10.3389/fvets.2020.00203/full |
| work_keys_str_mv | AT maiaelizalde aduplexquantitativerealtimereversetranscriptionpcrforsimultaneousdetectionanddifferentiationofflavivirusesofthejapaneseencephalitisandntayaserocomplexesinbirds AT cristinacanogomez aduplexquantitativerealtimereversetranscriptionpcrforsimultaneousdetectionanddifferentiationofflavivirusesofthejapaneseencephalitisandntayaserocomplexesinbirds AT franciscollorente aduplexquantitativerealtimereversetranscriptionpcrforsimultaneousdetectionanddifferentiationofflavivirusesofthejapaneseencephalitisandntayaserocomplexesinbirds AT elisaperezramirez aduplexquantitativerealtimereversetranscriptionpcrforsimultaneousdetectionanddifferentiationofflavivirusesofthejapaneseencephalitisandntayaserocomplexesinbirds AT laiacasadesmarti aduplexquantitativerealtimereversetranscriptionpcrforsimultaneousdetectionanddifferentiationofflavivirusesofthejapaneseencephalitisandntayaserocomplexesinbirds AT pilaraguilerasepulveda aduplexquantitativerealtimereversetranscriptionpcrforsimultaneousdetectionanddifferentiationofflavivirusesofthejapaneseencephalitisandntayaserocomplexesinbirds AT franciscoruizfons aduplexquantitativerealtimereversetranscriptionpcrforsimultaneousdetectionanddifferentiationofflavivirusesofthejapaneseencephalitisandntayaserocomplexesinbirds AT miguelangeljimenezclavero aduplexquantitativerealtimereversetranscriptionpcrforsimultaneousdetectionanddifferentiationofflavivirusesofthejapaneseencephalitisandntayaserocomplexesinbirds AT miguelangeljimenezclavero aduplexquantitativerealtimereversetranscriptionpcrforsimultaneousdetectionanddifferentiationofflavivirusesofthejapaneseencephalitisandntayaserocomplexesinbirds AT jovitafernandezpinero aduplexquantitativerealtimereversetranscriptionpcrforsimultaneousdetectionanddifferentiationofflavivirusesofthejapaneseencephalitisandntayaserocomplexesinbirds AT maiaelizalde duplexquantitativerealtimereversetranscriptionpcrforsimultaneousdetectionanddifferentiationofflavivirusesofthejapaneseencephalitisandntayaserocomplexesinbirds AT cristinacanogomez duplexquantitativerealtimereversetranscriptionpcrforsimultaneousdetectionanddifferentiationofflavivirusesofthejapaneseencephalitisandntayaserocomplexesinbirds AT franciscollorente duplexquantitativerealtimereversetranscriptionpcrforsimultaneousdetectionanddifferentiationofflavivirusesofthejapaneseencephalitisandntayaserocomplexesinbirds AT elisaperezramirez duplexquantitativerealtimereversetranscriptionpcrforsimultaneousdetectionanddifferentiationofflavivirusesofthejapaneseencephalitisandntayaserocomplexesinbirds AT laiacasadesmarti duplexquantitativerealtimereversetranscriptionpcrforsimultaneousdetectionanddifferentiationofflavivirusesofthejapaneseencephalitisandntayaserocomplexesinbirds AT pilaraguilerasepulveda duplexquantitativerealtimereversetranscriptionpcrforsimultaneousdetectionanddifferentiationofflavivirusesofthejapaneseencephalitisandntayaserocomplexesinbirds AT franciscoruizfons duplexquantitativerealtimereversetranscriptionpcrforsimultaneousdetectionanddifferentiationofflavivirusesofthejapaneseencephalitisandntayaserocomplexesinbirds AT miguelangeljimenezclavero duplexquantitativerealtimereversetranscriptionpcrforsimultaneousdetectionanddifferentiationofflavivirusesofthejapaneseencephalitisandntayaserocomplexesinbirds AT miguelangeljimenezclavero duplexquantitativerealtimereversetranscriptionpcrforsimultaneousdetectionanddifferentiationofflavivirusesofthejapaneseencephalitisandntayaserocomplexesinbirds AT jovitafernandezpinero duplexquantitativerealtimereversetranscriptionpcrforsimultaneousdetectionanddifferentiationofflavivirusesofthejapaneseencephalitisandntayaserocomplexesinbirds |