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

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S2 Abstracts / Journal of Clinical Virology 82S (2016) S1–S142
the scope of the Portuguese Influenza Surveillance Program, during
2014–2015 season in Portugal.
Methods: During the 2014–2015 season, 411 nasopharyngeal
swabs negative for influenzawere tested for HRV in amultiplex PCR
18% (75/411) of the samples were positive for HRV, and from
this 83% (62) were sequenced by a nested RT-PCR ucleotide
sequences of the VP4/VP2 region were used for genotyping and
phylogenetic tree construction in Mega 6.0. Demographic and clin-
ical data (according to EU ILI case definition) were recorded in a
questionnaire.
Results: HRV were detected throughout the study period,
between week 40/2014 (October) and week 9/2015 (March) with
a peak in January 2015. Phylogenetic analysis showed that 45%
(28/62) strains belonged to species HRV-A, 15% (9/62) to species
B and 40% (25/62) to species C. Overall were identified 35 different
types. All species co-circulated in Portugal with the exception of the
Algarve and Ac¸ ores, being HRV-A predominant in north region and
HRV-C predominant in Alentejo. HRV positives cases had a median
age of 42.5. HRV-C were the most frequently detected in all age
groups, apart from young adults aged 15 to 44. In this age group
HRV-A were identified in 73% (16/22). HRV-B was detected spo-
radically in all age groups, except in children (5-14 years old). HRV
was found in similar proportions in both genders (52% in female;
48% in male). Data on influenza vaccination was reported in 51
HRV positive cases, but only 9 (18%) had been previously vacci-
nated. Information on chronic diseases was reported in 54 cases, of
these 15 (28%) had a chronic disease (mainly cardiovascular or dia-
betes). HRV was detected in 4 pregnant women, 14% (4/28). Cough,
myalgias, weakness and fever were the most frequent symptoms
reported by HRV confirmed cases.
Conclusions: During 2014–2015 was observed a co-circulation
of the three species of HRV (A, B and C) with a predominance of
HRV-A followed by the recently identified species C. A wide genetic
diversity of 35 types was identified, with a higher diversity among
HRV-A. HRV was most frequently diagnosed in adults. Our study
included few children under 5, preventing conclusions about this
group. Diabetes and cardiovascular diseasewere found as a possible
risk for HRV infection, highlighting the relevance of respiratory dis-
ease prevention measures that should be undertaken. This was the
first study to attempt the genetic diversity of rhinovirus circulating
in Portugal during awinter season in ILI cases. Further studies in the
general population and in high-risk groups for severe respiratory
disease will aid knowledge in HRV epidemiology and exacerbation
of respiratory infections.
References
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[2] P. Linsuwanon, et al., J. Infect. 59 (2009) 115–121.
Abstract no: 224
Presentation at ESCV 2016: Oral 3
Illuminating influenza epidemiology in
Scotland using next generation sequencing
E. Goldstein ∗, P. Murcia , R. Gunson
1 West of Scotland Specialist Virology Centre,
Glasgow, Scotland, United Kingdom
2 University of Glasgow Centre for Virus Research,
Glasgow, Scotland, United Kingdom
Influenza is one of the most important respiratory pathogens
and is a major cause of mortality and morbidity worldwide every
year. Influenza A is a n RNA virus consisting of eight segments,
the segmented nature of the genome allows for reassortment to
occur, occasionally producing antigenically novel viruses capable
of causing influenza pandemics. The ECDC Scottish influenza lab-
oratory currently characterise influenza A isolates by sequencing
the HA1 region of the haemagglutinin gene, which allows isolates
to be classified into viral clades.
One hundred and fifty clinical isolates positive for influenza
A(H3N2) from the 2014/15 influenza season were sequenced by
both Sanger sequencing of HA1, in addition to whole genome
sequencing using next generation sequencing (NGS) technology on
the Illumina MiSeq platform. Influenza nucleic acid was amplified
using a single-reactionmethod, which simultaneously amplifies all
eight segments of the influenza genome. This amplified product
was then utilised for NGS.
Full segment coverage was achieved for the smaller segments
(NS and MP) of all 150 isolates, however coverage generally
decreased as the size of the segment increased. In total, 100%
genome coverage was achieved in 71 samples, with 100 samples
having >90% genome coverage. Sequencing of the haemagglutinin
gene was adequate for clade calling for all 150 isolates and phylog-
enies of the haemagglutinin gene constructed using NGS data had
better resolution than those produced using Sanger sequencing of
HA1 alone. In addition, using whole genome data we were able to
analyse isolates for evidence of viral reassortment and identified a
number of intra-clade reassortments in our dataset, involving both
the surface glycoproteins and internal genes. The majority of these
occurred sporadically, however one reassortant virus persisted in
the population.
Current routine influenza surveillance relying on sequencing
of the HA1 region allows for classification of influenza A viruses
into viral clades. Whole genome sequence data produced using a
single-reaction method and NGS allows for economical generation
of viral clade classification in addition to sequence data from the
other seven segments. In our small dataset we identified a number
of viral reassortments, suggesting that such events may occur more
often than previously estimated.
Abstract no: 195
Presentation at ESCV 2016: Oral 4
Long-term impairment attributable to
congenital cytomegalovirus infection
A.C.T.M. Vossen ∗, M.J. Korndewal ,
A.M. Oudesluys-Murphy , A.C.M. Kroes ,
M.A.B. van der Sande , H.E. de Melker
1 Leiden University Medical Center, Netherlands
2 Leiden University Medical Center, Centre for
Infectious Diseases, Epidemiology and Surveillance,
National Institute for Public Health and the
Environment, Netherlands
3 Centre for Infectious Diseases, Epidemiology and
Surveillance, National Institute for Public Health and
the Environment, Netherlands
Introduction: Congenital cytomegalovirus infection (cCMV) is
the most prevalent congenital infection worldwide and it may lead
to symptoms at birth as well as long term sequelae. Limited data
on long term sequelae are available, particularly in infants who are
asymptomatic at birth and in many studies on long-term conse-
quences a control group is lacking.
Aim and methods: A nation-wide retrospective cohort study
was designed to assess the long term consequences of cCMV up to