PCR-based next-generation West Nile virus sequencing protocols. (December 2021)

• Record Type: Journal Article
• Authors: Pappa, Styliani; Chaintoutis, Serafeim C.; Dovas, Chrysostomos I.; Papa, Anna
• Is Part Of: Molecular and cellular probes. Volume 60(2021)
• Publication Date: 2021-12
• Subjects: West nile virus -- Next generation sequencing -- Whole genome; Molecular probes -- Diagnostic use -- Periodicals; Pathology, Cellular -- Technique -- Periodicals; Cell Biology -- Periodicals; Molecular Biology -- Periodicals; Sondes moléculaires -- Utilisation diagnostique -- Périodiques; Cytopathologie -- Technique -- Périodiques; 572
• DOI: 10.1016/j.mcp.2021.101774
• Languages: English
• ISSNs: 0890-8508

Abstract

Abstract: The epidemiology of West Nile virus (WNV) is unpredictable and changing. Availability of whole genome sequences enables the detailed molecular epidemiology studies and the evaluation and design of diagnostic tools. In the present study we provide two PCR-based protocols which can be applied directly on biological samples from hosts infected by WNV strains belonging to lineage 1 or lineage 2. It was shown that the protocols worked successfully even on samples with relatively low viral load. Highlights: Two PCR-based NGS protocols were designed for whole genome sequencing of West Nile virus. The designed protocols can amplify WNV lineage 1 and both subclades of lineage 2 genomes. No prior culture amplification is required. The protocols work successfully even on samples with low viral load.