Journal of Clinical Virology - Volume 82 - Supplement 1 - September 2016

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Abstracts / Journal of Clinical Virology 82S (2016) S1–S142 S113
Abstract no: 182
Presentation at ESCV 2016: Poster 186
Prevalence and genetic characterization of
enterovirus D68 among children with severe
acute respiratory infection in China
W. Tan ∗, Y. Wang , Y. Zhao , R. Lu
1 National Institute for Disease Control and
Prevention, China
2 CDC, Beijing 102206, China
To understand the prevalence and molecular typing of
enterovirus D68 among children with severe acute respiratory
infection (SARI) in Beijing and Shanghai, 385 respiratory samples
were collected from in Beijing during 2008–2010, and 441 respi-
ratory samples were collected in Shanghai city between 2013 and
2014. All the samples were used for the screening of EV-D68 by nest
RT-PCR and sequencing, then EV-D68-positive samples were used
for the complete genome sequencing through overlapping PCR.
All available EV-D68 full-length genomes collected from GenBank
were used for phylogenetic analysis and comparison of EV-D68
types prevalent in China and America. One (0.4%) from 385 respira-
tory samples in Beijing was positive for EV-D68, and 4 (0.9%)among
the 441 samples from Shanghai were positive for EV-D68. Phylo-
genetic analysis of full length genome indicated that the EV-D68
prevalent in Beijing (BJ24) belong to Clade A2 and Clade B2, differ-
ent from the American popular strains (Clade A1, Clade B1, Clade
B4 and Clade B5). Partial sequence analysis declared phylogenetic
conflict among different gene sequences. We concluded that the
prevalence rate of EV-D68 among SARI Children in Beijing and
Shanghai currently was lower (5/700; <1%), and the EV-D68 geno-
type prevalent in China and America belong to different clusters.
Partial sequence analysis indicated that intratypic recombinant
events may occur in EV-D68 prevalent in China.
Abstract no: 186
Presentation at ESCV 2016: Poster 187
Replication and immune response in HAE of
HCoV-HKU1 isolate from a pediatric patient
with severe acute respiratory infection
N. Zhu ∗, R.J. Lu, W.J. Tan
National Institute for Viral Disease Control and
Prevention, Chinese Center for Disease Control and
Prevention, Beijing, China
Human coronavirus HKU1 (HCoV-HKU1), a fastidious cultured
-coronavirus, was associated with acute respiratory infection
in the aged and children. Human airway epithelium cells (HAE)
provide the first line of defense in the respiratory tract and are
the main target of HCoV-HKU1. However, little attention has been
devoted to immune response of HAE induced by HCoV-HKU1,
maybe due to its fastidious culturing. Here, we isolated a novel
strain of HCoV-HKU1 (BJ-01) from a pediatric patient with severe
acute respiratory infection (SARI) and propagated on HAE. This
stain of virus owned the typical morphology of coronavirus with
the diameter of 120–130 nm. The genome HCoV-HKU1 BJ-01 is
conserved during serially passage on HAE cells. Comparing viral
genome of the early passage (P3, Gene accession No KT779555)
with that of late passage (P9, Gene accession No KT779556), there
were only two amino acid substitutions on ORF1b (T2346C) and S
(G23216T) glycoprotein. We further investigate how the immune
response in HAE to HCoV-HKU1 infection using Quantibody®
Human Cytokine Antibody Arrays (RayBiotech, Inc.). We found
31 cytokines increased and 55 cytokines decreased more than
2 folds in the 640 detected cytokines. These cytokines can be
divided into different groups: Chemokines (CCL4, CCL13, CCL15,
CCL16, CCL24, CCL26, CXCL13, XCL1), Hematopoietins (IL23R, TSLP,
PRL, GHR), PDGF family (PDGFC, KITLG), IL-10 family (IL20RA),
IL-1 family (IL1R1), TNF family (SF11B, LTA, SF1B), TGF- fam-
ily (BMP7, BMPR1B), which mainly affect Chemokine/NF-KAPPA
B/PI3K-AKT/JAK-STAT signaling pathway on HAE cells. This work
was the first report on immune response in HAE of a novel HCoV-
HKU1 strain (HCoV-HKU1 BJ01) from a pediatric patient with SARI.
Abstract no: 19
Presentation at ESCV 2016: Poster 188
Development of an external quality assessment
panel for the molecular detection of respiratory
viruses
E. Elenoglou∗, F. Kartal, B. Kele, H. Seyedzadeh,
P. Jovanovic, S. Rughooputh, C. Walton
Public Health England, United Kingdom
Background: Respiratory virus infections occur commonly and
are responsible for a significant amount of morbidity worldwide. In
the developed world respiratory viruses are responsible for a con-
siderable amount of morbidity which has a significant economic
impact. Mortality rates however are low. In contrast, in develop-
ing countries, viruses are responsible for approximately 20–30% of
respiratory deaths in children. The spectrumof disease ranges from
upper respiratory tract infections such as common colds to infec-
tions of the lower respiratory tract manifesting as bronchiolitis or
pneumonia.
Respiratory Syncytial Virus (RSV) is the most common cause of
lower respiratory tract infection in infants and children worldwide.
Most frequent types of influenza viruses that affect individuals are
Influenza A and B, however rhinoviruses are the common cause of
coughs and colds during winter with a peak season in the UK to be
between January and March.
The elderly and infant population tend to be the most suscep-
tible to these viral illnesses. The availability of an External Quality
Assessment (EQA) panel for the Molecular Detection of Respiratory
viruses is crucial for providing objective evidence in the quality of
testing with a request.
Materials andmethods:A pilot distribution, containing 9 speci-
mens was sent out to 23 participants. Three simulated throat swabs
and six simulated freeze-dried nasopharyngeal aspirate material
(Specimen numbers were 3642–3650) were dispatched for testing
for respiratory pathogens usingmolecularmethods. The swab spec-
imens were distributed in a Phosphate Buffer Solution (PBS) based
solution with a different cell line added in each of them. The freeze
dried specimens were distributed in a sucrose based matrix. Sim-
ulated specimens were positive for Parainfluenza virus type one
(PIV-1), Adenovirus type 2 (AdV-2) and Influenza B (FluB).
Results: Out of 23 participants, 21 returned their results. Only
one laboratory detected PIV-1 in specimen 3642. PIV-1 was cor-
rectly reported for specimen 3643 mainly by those laboratories
who have used in-house real-time multiplex/single target PCR
assays. RespiFinder was the only commercially available multiplex
real-time assay that was able to detect PIV-1 in specimen 3643.
Adenovirus type 2 was successfully detected for specimens
3644, 3645 and 3649 by all those laboratories tested 100% for
adenoviruses. Performance was excellent for Flu-B with 100% of