Title
Phylogeographic diversity and hybrid zone of Hantaan orthohantavirus collected in Gangwon Province, Republic of Korea
Author(s)
Lee, Geum-Young;Kim, Won-Keun;Park, Kyungmin;Lee, Seung-Ho;Hwang, Jusun;et al.
Published
2020
Publisher
PLoS Neglected Tropical Diseases
Abstract
Background: Hantaan orthohantavirus (Hantaan virus, HTNV), harbored by Apodemus agrarius (the striped field mouse), causes hemorrhagic fever with renal syndrome (HFRS) in humans. Viral genome-based surveillance at new expansion sites to identify HFRS risks plays a critical role in tracking the infection source of orthohantavirus outbreak. In the Republic of Korea (ROK), most studies demonstrated the serological prevalence and genetic diversity of orthohantaviruses collected from HFRS patients or rodents in Gyeonggi Province. Gangwon Province is a HFRS-endemic area with a high incidence of patients and prevalence of infected rodents, ROK. However, the continued epidemiology and surveillance of orthohantavirus remain to be investigated. Methodology/Principal findings: Whole-genome sequencing of HTNV was accomplished in small mammals collected in Gangwon Province during 2015–2018 by multiplex PCR-based next-generation sequencing. To elucidate the geographic distribution and molecular diversity of viruses, we conducted phylogenetic analyses of HTNV tripartite genomes. We inferred the hybrid zone using cline analysis to estimate the geographic contact between two different HTNV lineages in the ROK. The graph incompatibility based reassortment finder performed reassortment analysis. A total of 12 HTNV genome sequences were completely obtained from A. agrarius newly collected in Gangwon Province. The phylogenetic and cline analyses demonstrated the genetic diversity and hybrid zone of HTNV in the ROK. Genetic exchange analysis suggested the possibility of reassortments in Cheorwon-gun, a highly HFRS endemic area. Conclusions/Significance: The prevalence and distribution of HTNV in HFRS-endemic areas of Gangwon Province enhanced the phylogeographic map for orthohantavirus outbreak monitoring in ROK. This study revealed the hybrid zone reflecting the genetic diversity and evolutionary dynamics of HTNV circulating in Gangwon Province. The results arise awareness of rodent-borne orthohantavirus diseases for physicians in the endemic area of ROK. Author summary: The genetic and molecular epidemiological studies on small mammals derived from hemorrhagic fever with renal syndrome (HFRS) endemic areas have consistently conducted for the public health surveillance and mitigation of orthohantavirus outbreak in the Republic of Korea (ROK). Implementing viral genome-based surveillance at new expansion sites that may identify HFRS risks is critical for tracking the location of orthohantavirus infections and diagnosing HFRS. In the present study, whole-genome sequences of Hantaan virus (HTNV) from small mammals in Gangwon Province were recovered using multiplex PCR-based next-generation sequencing during 2015–2018. In HFRS-endemic regions including Cheorwon-gun, Chuncheon-si, and Hwacheon-gun, additional HTNV genome sequences contribute to establish a high-resolution phylogeographic map for tracking emerging orthohantavirus-induced diseases. The cline analysis revealed a remarkable hybrid zone by showing spatial contact regions of HTNV at two sites (Cheorwon-gun and Hwacheon-gun) and the spatial separation and sequence divergence across genome segments of HTNV in Gangwon Province. These results demonstrate the genetic diversity and hybrid zone of HTNV circulating in Gangwon Province. These findings increase an awareness raising about HFRS in the endemic area of ROK.
Keywords
phylogeography;hemorrhagic fever with renal syndrome;genetic epidemiology;mammals;Polymerase chain reaction;rodents;microbial genetics;next-generation sequencing

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