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Complete Genome Sequence Analysis of Echovirus 18 Associated with

Aseptic Meningitis in Hebei Province, China, in 2015
Xiang-peng Chen,a Su-zhen Sun,b Jia-yun Guo,a Jing-jie Li,b Zheng-de Xiea
Key Laboratory of Major Diseases in Children, Ministry of Education, National Key Discipline of Pediatrics (Capital Medical University), National Clinical Research Center for
Respiratory Diseases, Beijing Key Laboratory of Pediatric Respiratory Infection Diseases, Beijing Pediatric Research Institute, Beijing Children’s Hospital, Capital Medical
University, Beijing, Chinaa; Children’s Hospital of Hebei Province, Shijiazhuang, Hebei Province, Chinab

Echovirus 18 is a member of the genus Enterovirus, family Picornaviridae, which can cause meningitis in children. Here, we re-
port the echovirus 18 complete genome sequence, which was isolated from the cerebrospinal fluid of a child with aseptic menin-
gitis in Hebei Province, China.

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Received 25 August 2016 Accepted 8 September 2016 Published 27 October 2016
Citation Chen X-P, Sun S-Z, Guo J-Y, Li J-J, Xie Z-D. 2016. Complete genome sequence analysis of echovirus 18 associated with aseptic meningitis in Hebei Province, China, in
2015. Genome Announc 4(5):e01165-16. doi:10.1128/genomeA.01165-16.
Copyright © 2016 Chen et al. This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International license.
Address correspondence to Zheng-de Xie, xiezhengde@bch.com.cn.

E chovirus 18 (Echo 18) is a member of the genus Enterovirus,

family Picornaviridae. It has been determined to be one of 63
serotypes of the human enterovirus B group, which usually cause
mild and self-limited disease and also can cause severe central ner-
vous system infection, such as acute flaccid paralysis (AFP), enceph-
alitis, and aseptic meningitis (1). Echo 18-related neurological dis-
eases have been reported in many countries and regions of the world
since it was first discovered in 1955 (2–6). However, to date, only 15
complete genome sequences of Echo 18 are available in GenBank
(prototype strain Metcalf, South Korean isolate Kor05-ECV18-
054cn, and 13 isolates from Germany submitted in April 2016).
In this study, the Echo 18 strain was isolated from a nine-year-
old boy clinically diagnosed with aseptic meningitis in Hebei
Province of China in 2015. The patient had acute onset of fever,
headache, nausea, and vomiting. A cerebrospinal fluid (CSF) sam-
ple was collected within 24 h after the onset of symptoms. The
pathogen initially tested positive for enterovirus by real-time re-
verse transcription-PCR (RT-PCR) and then was further identi-
fied as Echo 18 by RT-PCR and sequencing.
The clinical strain was isolated by culturing the CSF in human
rhabdomyosarcoma (RD) cell lines. After serial passage and
plaque purification, the recovered strain, named E18-314/HB/
CHN/2015, was obtained. Viral RNA was extracted with the
RNeasy minikit (Qiagen), cDNA was synthesized through
reverse-transcribed PCR, and a DNA library was prepared using
NEBNext Ultra DNA library prep kit for Illumina, according to
the manufacturer’s instructions, and sequenced using the MiSeq
platform (Illumina, PE 300). The next-generation sequencing
data were assembled using the CAP3 program. Phylogenic analy-
sis was performed by the MEGA6 program.
The complete genome of E18-314/HB/CHN/2015 contains
7,414 nucleotides, including a 5= untranslated region (5= UTR) of
743 nucleotides, a 3= untranslated region (3= UTR) of 101 nucle-
otides, and a single open reading frame (ORF) of 6,570 nucleo-
tides encoding a polyprotein of 2,190 amino acids. Comparing the
complete genomes of E18-314/HB/CHN/2015 with that of proto-
type strain Metcalf, the whole-genome nucleotide sequence iden-
tity is 80.6%, while the nucleotide sequence identities in the 5=
UTR, P1, P2, P3, 3= UTR, and VP1 regions are 81.5%, 80.2%,
79.7%, 81.0%, 87.1%, and 80.7%, respectively.
Phylogenetic analysis based on the VP1 sequences of Echo 18 in
GenBank showed that all the sequences of Echo 18 isolates from
Hebei Province are clustered into a distinct branch.
This report describes the first complete genome sequence of an
Echo 18 isolate in China, which belonged to human enterovirus B
species in the family Picornaviridae.
Accession number(s). The complete genomic sequence of
E18-314/HB/CHN/2015 was deposited in GenBank under the ac-
cession no. KX767786.
ACKNOWLEDGMENTS
This study was supported by National Major Science and Technology
Research Projects for the Control and Prevention of Major Infectious
Diseases in China (grant 2013ZX10004-202), Beijing Outstanding Young
Talent Training Projects (grant 2014000021469G237), and Capital
Clinical Feature Project of Beijing Technology Program (grant
Z151100004015046).
FUNDING INFORMATION
This work, including the efforts of Zheng-de Xie, was funded by
National Major Science and Technology Research Projects for the
Control and Prevention of Major Infectious Diseases in China
(2013ZX10004-202). This work, including the efforts of Xiang-peng
Chen, was funded by Beijing Outstanding Young Talent Training
Projects (2014000021469G237). This work, including the efforts of
Xiang-peng Chen and Zheng-de Xie, was funded by Capital Clinical
Feature Project of Beijing Technology Program (Z151100004015046).
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