First Detection of Enterovirus D68 in Korean Children, September 2022

Background: Enterovirus D68 (EV-D68) is a re-emerging pathogen that is particularly common in children and may cause asthma-like respiratory infection and acute flaccid myelitis. However, in Korea, EV-D68 has never been reported thus far. This study aimed to identify EV-D68 from nasopharyngeal aspirates (NPAs) in Korean children with a respiratory tract infection. Materials and Methods: The EV-D68 reference strain was purchased and blindly used to assess the detection ability of three commercial and one in-house mRT-PCR kit in 2018. Then, we selected children whose specimens were positive for human rhinovirus (HRV) and/or enterovirus (EV) by Allplex mRT-PCR (Seegene, Inc., Seoul, Korea) from April to December 2022. Total RNA was extracted from NPAs, and a partial 5'-UTR gene was amplified and sequenced for the identification of HRV/EV species. Additionally, PCR targeting the VP1 gene was performed to assess EV-D68positive NPAs, followed by sequencing. Phylogenetic analysis and comparison of amino acid sequence alignments were performed using a partial VP1 gene of our and recent international EV-D68 strains. Results: Among the mRT-PCR kits tested, only the in-house kit was able to detect EV-D68 in 2018. However, we detected three EV-D68 strains among children hospitalized with fever and/or respiratory symptoms in September - December 2022 who tested positive for EV by the Allplex kit. Two of them were healthy toddlers with lower respiratory infections accompanied by new-onset wheezing but no neurologic complications. Among 34 children with lower respiratory infection who tested positive for HRV during the same period, EV-D68 was not detected. Phylogenetic analysis revealed that the first Korean EV-D68 belonged to subclade B3. Amino acid sequence alignment of international subclade B3 EV-D68 strains also showed that our strain is genetically more related to Conclusion: We first detected EV-D68 in three Korean children who had EV detected by the Allplex mRT-PCR kit in 2022. EV-D68 also circulated in Korea in fall 2022, but the prevalence and severity seemed to be lower than those in previous reports from other countries.