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Detection of Astrovirus in a Cow with Neurological Signs by Nanopore Technology, Italy

In this study, starting from nucleic acids purified from the brain tissue, Nanopore technology was used to identify the etiological agent of severe neurological signs observed in a cow which was immediately slaughtered. Histological examination revealed acute non-suppurative encephalomyelitis affect...

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Autores principales: Zaccaria, Guendalina, Lorusso, Alessio, Hierweger, Melanie M., Malatesta, Daniela, Defourny, Sabrina VP, Ruggeri, Franco, Cammà, Cesare, Ricci, Pasquale, Di Domenico, Marco, Rinaldi, Antonio, Decaro, Nicola, D’Alterio, Nicola, Petrini, Antonio, Seuberlich, Torsten, Marcacci, Maurilia
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7290991/
https://www.ncbi.nlm.nih.gov/pubmed/32403368
http://dx.doi.org/10.3390/v12050530
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author Zaccaria, Guendalina
Lorusso, Alessio
Hierweger, Melanie M.
Malatesta, Daniela
Defourny, Sabrina VP
Ruggeri, Franco
Cammà, Cesare
Ricci, Pasquale
Di Domenico, Marco
Rinaldi, Antonio
Decaro, Nicola
D’Alterio, Nicola
Petrini, Antonio
Seuberlich, Torsten
Marcacci, Maurilia
author_facet Zaccaria, Guendalina
Lorusso, Alessio
Hierweger, Melanie M.
Malatesta, Daniela
Defourny, Sabrina VP
Ruggeri, Franco
Cammà, Cesare
Ricci, Pasquale
Di Domenico, Marco
Rinaldi, Antonio
Decaro, Nicola
D’Alterio, Nicola
Petrini, Antonio
Seuberlich, Torsten
Marcacci, Maurilia
author_sort Zaccaria, Guendalina
collection PubMed
description In this study, starting from nucleic acids purified from the brain tissue, Nanopore technology was used to identify the etiological agent of severe neurological signs observed in a cow which was immediately slaughtered. Histological examination revealed acute non-suppurative encephalomyelitis affecting the brainstem, cerebrum, cerebellum, and medulla oblongata, while by using PCR-based assays, the nucleic acids of major agents for neurological signs were not detected. By using Nanopore technology, 151 sequence reads were assigned to Bovine Astrovirus (BoAstV). Real-time RT-PCR and in situ hybridization (ISH) confirmed the presence of viral RNA in the brain. Moreover, using the combination of fluorescent ISH and immunofluorescence (IF) techniques, it was possible to detect BoAstV RNA and antigens in the same cells, suggesting the active replication of the virus in infected neurons. The nearly whole genome of the occurring strain (BoAstV PE3373/2019/Italy), obtained by Illumina NextSeq 500, showed the highest nucleotide sequence identity (94.11%) with BoAstV CH13/NeuroS1 26,730 strain, an encephalitis-associated bovine astrovirus. Here, we provide further evidence of the role of AstV as a neurotropic agent. Considering that in a high proportion of non-suppurative encephalitis cases, which are mostly indicative of a viral infection, the etiologic agent remains unknown, our result underscores the value and versatility of Nanopore technology for a rapid diagnosis when the PCR-based algorithm gives negative results.
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spelling pubmed-72909912020-06-17 Detection of Astrovirus in a Cow with Neurological Signs by Nanopore Technology, Italy Zaccaria, Guendalina Lorusso, Alessio Hierweger, Melanie M. Malatesta, Daniela Defourny, Sabrina VP Ruggeri, Franco Cammà, Cesare Ricci, Pasquale Di Domenico, Marco Rinaldi, Antonio Decaro, Nicola D’Alterio, Nicola Petrini, Antonio Seuberlich, Torsten Marcacci, Maurilia Viruses Article In this study, starting from nucleic acids purified from the brain tissue, Nanopore technology was used to identify the etiological agent of severe neurological signs observed in a cow which was immediately slaughtered. Histological examination revealed acute non-suppurative encephalomyelitis affecting the brainstem, cerebrum, cerebellum, and medulla oblongata, while by using PCR-based assays, the nucleic acids of major agents for neurological signs were not detected. By using Nanopore technology, 151 sequence reads were assigned to Bovine Astrovirus (BoAstV). Real-time RT-PCR and in situ hybridization (ISH) confirmed the presence of viral RNA in the brain. Moreover, using the combination of fluorescent ISH and immunofluorescence (IF) techniques, it was possible to detect BoAstV RNA and antigens in the same cells, suggesting the active replication of the virus in infected neurons. The nearly whole genome of the occurring strain (BoAstV PE3373/2019/Italy), obtained by Illumina NextSeq 500, showed the highest nucleotide sequence identity (94.11%) with BoAstV CH13/NeuroS1 26,730 strain, an encephalitis-associated bovine astrovirus. Here, we provide further evidence of the role of AstV as a neurotropic agent. Considering that in a high proportion of non-suppurative encephalitis cases, which are mostly indicative of a viral infection, the etiologic agent remains unknown, our result underscores the value and versatility of Nanopore technology for a rapid diagnosis when the PCR-based algorithm gives negative results. MDPI 2020-05-11 /pmc/articles/PMC7290991/ /pubmed/32403368 http://dx.doi.org/10.3390/v12050530 Text en © 2020 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Zaccaria, Guendalina
Lorusso, Alessio
Hierweger, Melanie M.
Malatesta, Daniela
Defourny, Sabrina VP
Ruggeri, Franco
Cammà, Cesare
Ricci, Pasquale
Di Domenico, Marco
Rinaldi, Antonio
Decaro, Nicola
D’Alterio, Nicola
Petrini, Antonio
Seuberlich, Torsten
Marcacci, Maurilia
Detection of Astrovirus in a Cow with Neurological Signs by Nanopore Technology, Italy
title Detection of Astrovirus in a Cow with Neurological Signs by Nanopore Technology, Italy
title_full Detection of Astrovirus in a Cow with Neurological Signs by Nanopore Technology, Italy
title_fullStr Detection of Astrovirus in a Cow with Neurological Signs by Nanopore Technology, Italy
title_full_unstemmed Detection of Astrovirus in a Cow with Neurological Signs by Nanopore Technology, Italy
title_short Detection of Astrovirus in a Cow with Neurological Signs by Nanopore Technology, Italy
title_sort detection of astrovirus in a cow with neurological signs by nanopore technology, italy
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7290991/
https://www.ncbi.nlm.nih.gov/pubmed/32403368
http://dx.doi.org/10.3390/v12050530
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