First detection of Lake Sinai virus in the Czech Republic: a potential member of a new species.

Lake Sinai virus (LSV) is one of over 20 honey bee viruses. Variants of LSV have been classified as members of two officially recognised species, Lake Sinai virus 1 and Lake Sinai virus 2. However, there are currently a limited number of whole-genome sequences, and the genetic variability of the virus indicates that additional species may need to be established. Extracted nucleic acid of 209 honey bee samples was screened by PCR for 11 honey bee viruses. LSV was the third most abundant virus (36.9% of positive samples), after Apis mellifera filamentous virus (72.2%) and deformed wing virus (52.5%). LSV-positive samples were analyzed further by PCR with primers targeting the region encoding the viral RNA-dependent RNA polymerase. Subsequently, the PCR products were sequenced, and the resulting sequences were used for a first round of phylogenetic analysis. Based on those results, several isolates were selected for whole-genome sequencing, and the complete genome sequences were used for additional phylogenetic analysis. The results indicated the presence of at least three genetically distinct groups of LSV in the Czech Republic, the most prevalent one being related to LSV 2 but too dissimilar to be considered a member of the same species. Two sequences of a major LSV strain cluster native to the Czech Republic were determined, representing the first Czech LSV strains published to date.