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Fig 1.

Geographic distribution of trapping sites for A. agrarius collected in Gangwon and Gyeonggi provinces, the Republic of Korea.

A map shows the trapping sites where A. agrarius was captured from 2003–2014. Color circles indicate the places where the positivity of HTNV was shown by RT-PCR; Dagmar North (DN), red; Twin Bridge Training Area North (TBTA-N), violet; TBTA South (TBTA-S), yellowish green; Nightmare Range (NR), green; Fire Point 131 (FP131), orange; Cheorwon, black; Hwacheon, cyan; opened circles indicate the places where no HTNV RNA was identified.

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Fig 1 Expand

Table 1.

Total number and screening results of A. agrarius collected from Gyeonggi and Gangwon provinces in the Republic of Korea, 2003–2014.

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Table 1 Expand

Fig 2.

The serological and molecular prevalence of HTNV from A. agrarius in Gangwon and Gyeonggi provinces.

A total of 5,929 A. agrarius were captured in the Gangwon and Gyeonggi provinces from 2003–2014. A total of 774 A. agrarius were seropositive for anti-HTNV IgG (shown by black bars). Grey bars represent 544/774 A. agrarius that were positive for the partial HTNV M segment. Arabic number above the bars indicates the number of IFA and RT-PCR positive rodents, respectively.

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Table 2.

Accession number of the whole genome sequences of HTNV collected from Gyeonggi and Gangwon provinces.

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Table 2 Expand

Fig 3.

Phylogenetic analysis of whole HTNV L segment genomic sequences in Gangwon and Gyeonggi provinces.

The full-length sequences of the HTNV L segment were obtained from lung tissues of A. agrarius in Gangwon and Gyeonggi provinces. A phylogenetic tree was generated by the ML method. Topologies were evaluated by bootstrap analyses of 1000 iterations. The TN93 (Tamura-Nei)+G model of evolution was used, based on an alignment of the entire nucleotide sequence of the L segment, including strains HTN 76–118 (X55901) and HTN HV004 (JQ083393).

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Fig 4.

Phylogenetic analysis of whole HTNV M segment genomic sequences in Gangwon and Gyeonggi provinces.

The full-length sequences of the HTNV M segment were obtained from lung tissues of A. agrarius in Gangwon and Gyeonggi provinces. A phylogenetic tree was generated by the ML method. Topologies were evaluated by bootstrap analyses of 1000 iterations. The T92 (Tamura 3-parameter)+G model of evolution was used, based on an alignment of the entire nucleotide sequence of the M segment, including strains HTN 76–118 (M14627) and HTN HV004 (JQ083394).

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Fig 5.

Phylogenetic analysis of whole HTNV S segment genomic sequences in Gangwon and Gyeonggi provinces.

The full-length sequences of the HTNV S segment were obtained from lung tissues of A. agrarius in Gangwon and Gyeonggi provinces. A phylogenetic tree was generated by the ML method. Topologies were evaluated by bootstrap analyses of 1000 iterations. The T92+G model of evolution was used, based on an alignment of the nucleotide sequence of the S segment, including strains HTN 76–118 (M14626) and HTN HV004 (JQ083395).

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Fig 6.

Evidence of a possible genetic reassortment event between HTNV strains in the ROK.

(A) p-values from the RDP program. (B) The Bootscan plot was based on a pairwise distance model by the RDP4 algorithm. A Bootscan Support Percent of over 70% (cutoff value) was considered significant. Cyan color represents for the comparison of the 7 group (DN, Paju) to the 6 group (Hwacheon); Violet color represents for the comparison of the 7 group (DN, Paju) to the 2 group (TBTA-N, Paju). (C) A reassortant, in-, and out-groups of HTNV were phylogenetically analyzed by the construction of individual ML trees for the L, M, and S segments. The reassortant (the 7 group) is shown in red color. The 6 group (Hwacheon) and the 2 group (TBTA-N, Paju) are indicated by cyan and violet colors, respectively.

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