Enterovirus D68 (EV-D68) is a respiratory virus that causes infections of the upper and lower respiratory tract and can also be associated with various central nervous system complications, including acute flaccid myelitis (AFM). The aim of this study was ro assess the incidence, seasonality, and molecular epidemiology of EV-D68 in children in Slovenia, and to describe the clinical characteristics of children infected with EV-D68 between 2014 and 2022. We included children who were hospitalized or examined as outpatients in the University Medical Center Ljubljana. EV-D68 was detected by specific rtRT-PCR. The analysis revealed a biennial pattern of EV-D68 circulation, with peaks in infections occurring in late summer and early autumn in 2014 and 2016. Sanger sequencing and the CLC Main Workbench version 7 and MEGA X software were used to determine the molecular epidemiology. In Slovenia, both subclades B1 and B2 were present simultaneously in 2014, while in the following years, we only identified EV-D68 belonging to subclade B3. Severe respiratory infections and the potential neurotropism of the virus indicate the importance of monitoring EV-D68.
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