Whole genome sequencing and genetic characterization of Dobrava-Belgrade Orhohantavirus isolate from Kırklareli province with next generation sequencing

Ortohantaviruses are transmitted to humans mostly through small mammals, which are also carriers of these viruses. Because orthohantaviruses show great genetic variability through regions, it is of great importance to determine of genetic characterization with whole genome sequencing. Dobrava-Belgrade orthohantavirus (DOBV) positivity has been shown in 8 rodents which were from İğneada region, Kırklareli province, with the previous work of our team. These positivity has been detected by performing PCR and Sanger sequencing from the lung tissues of the rodents. The aim of this study is the whole genome sequencing of all three segments of DOBV. Therefore, rRNA depletion kit and RNA library preparation kit were used to prepare the libraries for whole genome sequencing. After that, Illumina MiSeq system was used for sequencing of libraries. Finally, phylogenetic trees of the sequences were constructed with the sequences of different DOBV strains to determine the differences of DOBV İğneada strain. As a result, whole genome sequences of DOBV were obtained from Turkey for the very first time. The complete sequences of all three segments were obtained directly from lung tissue and thus, complete L segment sequence became the only sequence that has this unique property for DOBV on NCBI GenBank database. The phylogenetic analysis has shown that there is a close relation between DOBV Igneada strain from Kırklareli province from Turkey and DOBV Ano-Poroia strain from Greece. SimPlot analysis has shown the similarities between DOBV Igneada strain and other DOBV strains from the Balkans.