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

Viruses

Nicola Decaro¹  Torsten Seuberlich²  SabrinaVP Defourny³  Nicola D’Alterio³  Maurilia Marcacci³  Guendalina Zaccaria³  Antonio Rinaldi³  Daniela Malatesta³  MarcoDi Domenico³  Alessio Lorusso³  Cesare Cammà³  Antonio Petrini³  MelanieM. Hierweger⁴  Franco Ruggeri⁵  Pasquale Ricci⁵ 

[1] Dipartimento di Medicina Veterinaria, Università degli Studi di Bari, 70010 Valenzano, Italy;Graduate School for Cellular and Biomedical Sciences, University of Bern, 3012 Bern, Switzerland;Istituto Zooprofilattico Sperimentale dell’Abruzzo e Molise, 64100 Teramo, Italy;NeuroCenter, Department of Clinical Research and Veterinary Public Health, University of Bern, 3012 Bern, Switzerland;Unità Operativa Complessa Servizio di Sanità Animale, ASL Pescara, 65100 Pescara, Italy;

关键词: bovine astrovirus;    nanopore technology;    brain;    neurological signs;    cattle;    phylogeny;   

DOI  :  10.3390/v12050530

来源: DOAJ

【 摘 要 】

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.

【 授权许可】

Unknown