Bridging the Gap: Challenges and Innovations in Validating Next-Generation Sequencing Discoveries of Arboviruses

Next-generation sequencing (NGS) has dramatically accelerated the identification of viral genomic sequences from arthropod vectors, uncovering an unprecedented diversity of potential arboviruses. Nevertheless, the majority of these novel viral genomic sequences (NVGS) remain uncharacterised genomic data without corresponding viral isolates, highlighting the discrepancy between sequence detection and biological validation. This gap impedes our understanding of the biological properties, host range, and pathogenic potential of these newly identified viruses. To bridge this gap, the review advocates for integrated strategies, including expanded cell culture screening, novel cell line development, and reverse genetics approaches. These are complemented by artificial intelligence (AI) for predictive viral characterization. In parallel, NGS can be combined with other sequencing strategies to obtain more complete and accurate viral whole-genome sequences. Validating NGS discoveries is essential for effective risk assessment and outbreak preparedness against emerging arboviral threats, with important implications for public health surveillance.