Characterization of two novel Negevirus strains and analysis of global Negevirus distribution and host diversity

We report the first isolation of two Negevirus strains from Aedes and Culex mosquitoes in Xinjiang, China: Dezidougou virus (DEZV, isolate XJ-ALT23-420-01) and Negev-like virus (NEGLV, isolate XJ-JH20-91-01). Phylogenetically, the nucleotide sequence of DEZV exhibits high similarity (98.96%) with the DEZV isolate 8345 from Germany but shows lower similarity (<91.91%) with other DEZV strains. In contrast, the NEGLV isolate XJ-JH20-91-01 displays significant divergence from other Negevirus (NEGV) or NEGLV, sharing only 79.39% nucleotide similarity with the most closely related strain, NEGV BeAr805514. Both DEZV/XJ-ALT23-420-01 and NEGLV/XJ-JH20-91-01 replicated rapidly in mosquito cell lines (C6/36 and Aag2), reaching viral loads of up to 109-10 copies/mL, causing significant cytopathic effect (CPE) in these cell lines but failing to replicate in vertebrate cells.