Frontiers in cellular and infection microbiology · 2026
Combined NGS and proteomics improves viral diagnostics in wildlife: coronaviruses confirmed in hedgehogs.
СтатьяНазвание и аннотация: Перевод в очереди
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Chudinov Ivan, Korneenko Elena, Rog Irina, Lukina-Gronskaya Alexandra, Kovalenko Alexey, Gremyacheva Veronika, Butenko Ivan, Vasileva Elena, Matyushkina Daria, Litvinova Elena, Feoktistova Natalia, Speranskaya Anna
Аннотация
INTRODUCTION: Metaviromic sequencing is prone to artifacts, necessitating orthogonal validation. This study evaluates a targeted proteogenomic approach for verifying viral presence initially detected by NGS.
METHODS: We analyzed oropharyngeal/anal swabs and feces from hedgehogs, where NGS indicated viruses from the Coronaviridae, Picornaviridae, Flaviviridae, and Astroviridae families. For verification, mass spectrometry was applied using sample-specific databases derived from NGS-assembled contigs.
RESULTS: Proteomic analysis confirmed Coronaviridae presence in NGS-positive animals, primarily detecting structural virion proteins in anal and fecal samples. However, proteomic evidence for Picornaviridae, Flaviviridae, and Astroviridae was not obtained despite their genomic detection.
DISCUSSION: Phylogenetic analysis of two complete Picornaviridae genomes assembled from the NGS data - one closely related to a UK rodent virus and another distantly related to Chinese bat viruses - supports the authenticity of the NGS findings. The results demonstrate that this two-stage pipeline provides critical orthogonal validation for metaviromic studies but also defines a practical sensitivity limit for the proteomic verification of some viral families. Combining NGS with proteomics significantly enhances the reliability of viral confirmation in complex samples, thereby strengthening ecological and zoonotic inferences.
METHODS: We analyzed oropharyngeal/anal swabs and feces from hedgehogs, where NGS indicated viruses from the Coronaviridae, Picornaviridae, Flaviviridae, and Astroviridae families. For verification, mass spectrometry was applied using sample-specific databases derived from NGS-assembled contigs.
RESULTS: Proteomic analysis confirmed Coronaviridae presence in NGS-positive animals, primarily detecting structural virion proteins in anal and fecal samples. However, proteomic evidence for Picornaviridae, Flaviviridae, and Astroviridae was not obtained despite their genomic detection.
DISCUSSION: Phylogenetic analysis of two complete Picornaviridae genomes assembled from the NGS data - one closely related to a UK rodent virus and another distantly related to Chinese bat viruses - supports the authenticity of the NGS findings. The results demonstrate that this two-stage pipeline provides critical orthogonal validation for metaviromic studies but also defines a practical sensitivity limit for the proteomic verification of some viral families. Combining NGS with proteomics significantly enhances the reliability of viral confirmation in complex samples, thereby strengthening ecological and zoonotic inferences.