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

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S130 Abstracts / Journal of Clinical Virology 82S (2016) S1–S142
and Immunology, Faculty of Medicine, University of Ljubljana,
for the routine detection of respiratory viruses, including respi-
ratory syncytial virus (RSV), human rhinoviruses (hRV), human
metapneumovirus (hMPV), human coronaviruses (HCoVs), human
bocavirus (HBoV), adenoviruses (AdV), parainfluenza virus (PIV)
and influenza viruses A and B (Flu A-B) by real-time RT-PCR. HCoVs
positive samples with high viral load (low Ct value) and those
negative for all other respiratory viruses were include into fur-
ther testing by amplifying a 440-bp-long fragment of the highly
conserved polymerase gene.
Results: From December 2013 to February 2016, a total 16686
nasopharyngeal swabs from patients with acute respiratory tract
infections were enrolled in the study. From these 976 (5.8%) were
positive for HCoVs and 523 (58.6%) were negative for RSV, hRV,
hMPV, HCoVs, HBoV, AdV, PIV, Flu A and FluB by real-time RT-
PCR. From 523 HCoVs positive sample 129 were further tested
for all HCoVs species, including 47 HCoV-HKU1, 44 HCoV-OC43,
24 HCoV-NL63, 11 HCoV-229E, 1 HCoV-HKU1/HCoV-229E and
1 HCoV-NL63/HCoV-229E. Only HCoVs positive samples (HCoV-
HKU1 and HCoV-OC43) with high viral load (Ct-value less than
30) were include into further testing. To characterize the over-
all diversity of coronavirus sequences, 65 sequences have been
included in phylogenetic analysis; 31 sequences of HCoV-OC43 and
34 sequences of HCoV-HKU1.
Conclusions: Among four circulating HCoVs, HCoV-HKU1 and
HCoV-OC43 seem to show the highest prevalence and incidence in
hospitalized patients. The phylogenetic analysis shows that Slove-
nian human coronavirus strains from this study belong to the
four clusters, two grouping HCoV-OC43 and two HCoV-HKU1. The
present study draws genetically diversity of human coronaviruses
in Slovenian hospitalized patients.
Abstract no: 78
Presentation at ESCV 2016: Poster 221
Rapid diagnosis of respiratory viral infections in
primary health care
A.H.L. Bruning ∗ , W.B. de Kruijf ,
H.C.P.M. van Weert L.M. Willems
M.D. de Jong , D. Pajkrt , K.C. Wolthers
1 Department of Pediatric Infectious Diseases and
Immunology, Emma Children’s Hospital, AMC,
Amsterdam, The Netherlands
2 Department of General Practice, AMC, Amsterdam,
The Netherlands
3 Health Center Gein, GAZO, Amsterdam, The
Netherlands
4 Laboratory of Clinical Virology, Department of
Medical Microbiology, AMC, Amsterdam, The
Netherlands
Background: Respiratory tract infections (RTI) are the most
common acute problems in primary health care. RTIs are mainly
of viral origin. The epidemiology of respiratory viruses in primary
health care settings is scarcely reported, as diagnostic tests for RTIs
are sporadically used by general practitioners (GP). Rapid, sensi-
tive and specific identification of viral RTIs might assist diagnostic
interpretation and potentially prevent inappropriate use of antibi-
otics.
Aim: To increase our insight in the epidemiology of viral RTIs
in primary health care; to evaluate the feasibility and diagnos-
tic accuracy of a new rapid test for respiratory viruses (mariPOC®
test system, ArcDia International, Turku, Finland) in primary health
care.
Methods: Patients with RTI symptoms presenting to a primary
healthcare practice in the neighborhood of the Academic Medical
Center (AMC) Amsterdamwere asked to complete a small question-
naire about his/her symptoms and undergo nasopharyngeal swab
sampling. The swab was immediately tested at the point-of-care
with the automatedmariPOC® test. ThemariPOC ® test is a simple to
perform test for the detection of nine respiratory viruses (influenza
A and B, parainfluenza type 1, 2 and 3 viruses, respiratory syncy-
tial virus (RSV), human adenovirus, human bocavirus, and human
metapneumovirus) and Streptococcus pneumoniae, with prelimi-
nary results ready within 20min and final results within 2 h. The
remaining sample solution was transferred on the same day to the
Laboratory of Clinical Virology at the AMC for reference testingwith
multiplex PCR. Clinical and epidemiological data were collected
including age, gender, underlying illness, presenting symptoms,
time from onset of symptoms and detected viruses. The sensitivity
and specificity of themariPOC® as compared to PCRwas calculated.
The clinical feasibility of the mariPOC® test was evaluated using a
questionnaire for the study participants and GPs.
Results: From November 11 2015 till March 30 2016 a total of
371 patients (59.3% female, median age 45 years) were included.
One or more respiratory viruses were detected by PCR in 43.4%
(n = 161) of the collected nasopharyngeal swabs. Rhinovirus (RV)
was the most frequently detected virus with a prevalence of 11.9%.
When reporting sampleswithCt up to 40 as positive findings in PCR,
the sensitivity and specificity of the mariPOC® test were respec-
tively for influenza A virus ( n = 24), 54.2% and 98.9%; for influenza
B virus (n = 18), 72.2% and 99.5% and for RSV (n = 12), 50.0% and
100%. In samples with higher viral load (i.e. Ct-value < 30) sensi-
tivity for influenza A, influenza B and RSV was 85.7%, 78.6%, and
87.5%, respectively. The availability of a diagnostic test for respira-
tory viruses in primary healthcarewas appreciated by both patients
and GPs.
Conclusion: Respiratory viruses are frequent causes of RTIs in
primary health care. Acute infections with high viral loads were
accurately detected by the mariPOC test and for these infections a
rapid test would be a helpful tool for GPs. Both doctors and patients
were positive about the availability of a rapid test in primary health
care. The development of a rapid test for rhinovirus would be valu-
able as rhinovirus was the most frequently detected virus.
Abstract no: 82
Presentation at ESCV 2016: Poster 222
False-negative detection of respiratory syncytial
virus as an example that regular update of
RT-PCR is required for reliable molecular
detection of respiratory viruses
Els Wessels ∗, Roel Nijhuis, Jutte de Vries,
Eric Claas
Leiden University Medical Center, The Netherlands
Objectives: A respiratory sample that was RT-PCR adenovirus
positive and negative for other tested respiratory viruses was cul-
tured for adenovirus serotyping in December 2013. Surprisingly,
shell vial culture was positive for respiratory syncytial virus (RSV).
Methods: In June 2013 an update of the RT-PCR that was used
to detect respiratory viruses was started he update included
the following steps: updating alignments of every target with
sequences retrieved from GenBank, amplification and sequence