Detection of Astrovirus in a Cow with Neurological Signs by Nanopore Technology, Italy.
Guendalina ZaccariaAlessio LorussoMelanie M HierwegerDaniela MalatestaSabrina Vp DefournyFranco RuggeriCesare CammaPasquale RicciMarco Di DomenicoAntonio RinaldiNicola DecaroNicola D'AlterioAntonio PetriniTorsten SeuberlichMaurilia MarcacciPublished in: Viruses (2020)
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.
Keyphrases
- single molecule
- cerebral ischemia
- real time pcr
- solid state
- resting state
- induced apoptosis
- white matter
- loop mediated isothermal amplification
- machine learning
- spinal cord
- sars cov
- drug induced
- functional connectivity
- living cells
- liver failure
- high throughput
- label free
- quantum dots
- multiple sclerosis
- oxidative stress
- early onset
- spinal cord injury
- intensive care unit
- single cell
- respiratory failure
- dendritic cells
- brain injury
- nucleic acid
- room temperature
- hepatitis b virus
- blood brain barrier
- cell proliferation
- signaling pathway
- aortic dissection
- neural network