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

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Abstracts / Journal of Clinical Virology 82S (2016) S1–S142 S119
instruction by study nurses. Nucleic acids were extracted from
400 l of sample using NucliSENS easyMAG (bioMérieux, Marcy
l’Etoile, France). RNA and DNA were eluted in 110 l and ana-
lyzed by real-time PCR using a combination of 7 duplex Respiratory
Multi Well System r-geneTM assays (influenza A/B, RSV/hMPV,
Rhino&EV/cell control, ADV/HBoV, HCoV/HPIV1-4, Chla/Myco and
PeV) (Argene/bioMerieux,Marcy l’Etoile, France), according to the
manufacturer’s instructions. Sample quality was evaluated using a
HPRT1 cellular gene control (CC) assay (included in the Rhino&EV
assay) that evaluated the quantity of human epithelial cells present
in the sample.
Results: CC was positive in 93% of the samples (1294/1398) and
among those, 98% and 87% were positive in healthy and CF infant
groups, respectively. Semiquantitative analysis of positive CCs and
virus positive samples did not differ between the two groups. Anal-
yses of the CT values (with and without inclusion of low quality
swabs) did not demonstrate any differences between both study
groups. Rates of viral colonization were similar in healthy and CF
infants (43% and 42%, respectively), however there were clear dif-
ferences in PIV colonization (11% and 6%, respectively; p = 0.038)
and bocavirus colonization (6% and 23%, respectively; p < 0.001).
HRV was the most frequent virus identified in healthy and CF
infants (57% and 46%, respectively).
Conclusion: This study demonstrated that parental collection
of nasal swabs from healthy and CF infants provided easy and
adequate material for testing. Although the number of low qual-
ity swabs was slightly higher in the CF group, sensitivity analysis
showed that this did not bias the results. Possible reasons for lower
quality may be more careful swabbing by parents of infants with
CF or viscous mucus in the nose. Interestingly, while viral coloniza-
tion in general was similar in healthy and CF infants, there were
clear differences in viral species, a finding of importance for future
treatment options and understanding disease development.
Abstract no: 269
Presentation at ESCV 2016: Poster 199
Genotyping of rhinoviruses in children and
adults during 2014–2016
N.E. Demirkan ∗, S. Kirdar , E. Ceylan ,
A. Yenigün , I. Kurt Omurlu
1 Department of Medical Microbiology, Adnan
Menderes University, School of Medicine, Aydın,
Turkey
2 Pulmonary Medicine, Adnan Menderes University,
School of Medicine, Aydın, Turkey
3 Pediatric Diseases, Adnan Menderes University,
School of Medicine, Aydın, Turkey
4 Department of Biostatistics, Adnan Menderes
University, School of Medicine, Aydın, Turkey
Background and objective: Rhinoviruses (RV) are major
causative agents of acute respiratory tract infections in children
and adults. Rhinoviruses can be divided into three species (RV-A,
RV-B, and RV-C) including many different viral types on the basis
of their genetic characteristics. The objective of the study was to
determine the distribution of RV species in respiratory infections
between January 2014 and 2016 inAydın, Turkey, and to investigate
the relationship between the RV species and clinical symptoms.
Material andmethods: Between January 2014 and 2016, a total
127 nasopharyngeal swabs samples were collected from patients
submitted to AdnanMenderes UniversityHospital inAydın, Turkey.
The screening of respiratory viruses (HAdV, FLUAV, FLUBV, RV,
HCoV- 229E/NL63, HCoV-OC43, HMPV, HPIV-1, HPIV-2, HPIV-3,
HPIV-4, HEV, HRSVA, HRSV-B and HBoV) was performed with two
commercial multiplex PCR-basedmethod (AnyplexII RV16, Seegene,
South Korea and FTD Respiratory pathogens 21 plus, Luxemburg).
The RV-positive samples were sequenced in the VP4/VP2 regions.
Results: Of the 127 samples, 96 (75.5%) were positive (50 chil-
dren, 46 adults). The median age for children was 24 months
(6.19–72) and the mean age for adults was 57.63± 15.68. From
a total of 96 rhinovirus-positive samples, 65 (33 children and
32 adults) were sequenced in the VP4/VP2 regions. Twenty-eight
(43.07%) samples were identified as RV-A and 7 (10.76%) as RV-B,
and 28 (43.07%) samples belonging to the RV-C species. EV-D68
was detected in only one adult patient.
Conclusion: To our information, this is the first study about
RV genotyping in children and adult patients in Turkey. We have
detectedRV-A andRV-C being themost prevalent species, andHRV-
B trailing behindNo significant relationship between the RV species
and clinical symptoms was observed.
Abstract no: 277
Presentation at ESCV 2016: Poster 200
A cost-efficient solution: Reagent comparison
guide for neuraminidase inhibition assay
J. Louro ∗, V. Correia ,
H. Rebelo-de-Andrade
1 Host-Pathogen Interaction Unit, Research Institute
for Medicines (iMed.ULisboa), Faculdade de
Farmácia da Universidade de Lisboa, Portugal
2 Departamento de Doenc¸ as Infecciosas, Instituto
Nacional de Saúde Dr. Ricardo Jorge IP, Lisboa,
Portugal
Background:The neuraminidase (NA) inhibitors (NAIs) encom-
pass three FDA-approved compounds: oseltamivir (Tamiflu® ),
zanamivir (Relenza® ) and peramivir (Rapivab® ); along with lan-
inamivir (Inavir® ) approved in Japan [Takashita et al., 2015]. The
potential for emergence and spread of NAI-resistant viruses and
the limited therapeutic options available reinforce the importance
of NAI susceptibility surveillance [Okomo-Adhiambo et al., 2014].
Phenotypic profiling of influenza virus (IV) susceptibility to
NAIs tested by neuraminidase inhibition assay (NIA) has been the
recommendedmethodology to use, allowing to determine the con-
centration of NAI required to inhibit 50% of the virus NA activity
(IC50) [WHO, 2012]. The central reagents used in this assay, specif-
ically theMUNANA substrate and the antiviral drug can be obtained
frommultiples pharmaceutical companies at a different price. Con-
sequently, there is the need of practical guidance regarding the
chosen reagent suppliers, which may have an effective outcome
on the reported inter-laboratory results. In this context, the evalu-
ation of available reagents is essential to determine the efficacy of
the alternative sources for MUNANA and NAIs and possibly provide
a cost-efficient alternative solution.
This study aimed to compare available alternative reagents for
NA activity assay (NAA) and NIA and to assess the phenotypic sus-
ceptibility profiles of IV from different (sub)types to the new NAIs
laninamivir and peramivir.
Methods: Twelve IV were selected for study: 6 virus isolates
from a reference panel (isirv-Antiviral Group); and 6 clinical speci-
mens positive for influenza virus. Phenotypic assay was performed
using an in-house MUNANA-based IC50 fluorescence assay [HPA,
2006].