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

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S16 Abstracts / Journal of Clinical Virology 82S (2016) S1–S142
Results: We obtained 136 Megabases, with 94.5% of reads pass-
ing the error filtering algorithms (Nasu et al. PLoS ONE, 2011),
and 92% mapping to the regions of interest, with similar cover-
age (mean; range = 7660; 759–18,729). The overall error rate for
the procedure was 0.319%, which is similar to that obtained previ-
ously with the same HCV-NS3 protease clone in the 454 platform
(Salvatierra et al. Virologie, 2013). Using a majority rule consen-
sus sequence for each patient sample, several natural RAVs were
found at different prevalence. For NS3: A87V (2.3%), R117C/H (2.3%
each), S122N/T (2.3% each), V170I (25%) and S174A (2.3%). For
NS5A: R30Q (9.1%), L31I (3.3%), Y93H (4.5%). For NS5B: L159F
(22.7%), C316N (38.7%), C316H (2,3%), S368A (2.3%), Y448H (4.5%)
and S556G (15.91%). No RAVs were found in NS3-Q80, R155, A156,
D168, M175; NS5A-L28, P58; or NS5B-S282, S365, N411, M414,
G554 or D559.
Conclusions: The Illumina MiSeq platform allows for the simul-
taneous sequencing of HCV NS3, NS5A and NS5B regions, with
a high coverage and multiplexing capacity. Althoug clinically-
relevant variants may be represent frequencies >15% of the
quasispecies, after filtering the error rates obtained with the MiSeq
are similar to the Roche-454 platform, thus potentially allowing
for detecting minority variants down to 1% of the viral popula-
tion. While major resistance mutations to protease and nucleosidic
inhibitors are rare in our isolates, we detected the NS5A-R30Q and
Y93Hmajor RAVs, and the NS5B-L159F accessory and C316N RAVs.
Because the concept HCV treatment is being updated to finding
the right combination of DAAs in a particular patient, obtaining the
complete DAA susceptibility profile with this platform will be use-
ful to further treatment optimization and for the management of
DAA failures.
Abstract no: 107
Presentation at ESCV 2016: Oral 28
Hepatitis D virus infection in Slovenian patients
with chronic hepatitis B virus infection: A
national prevalence study
L. Hoˇsnjak ∗, M.M. Jelen , ˇS. ˇStunf , A. Zagoˇzen ,
K. Fujs Komloˇs arkoˇciˇc eme
M. Poljak
1 Institute of Microbiology and Immunology, Faculty
of Medicine, University of Ljubljana, Ljubljana,
Slovenia
2 Institute of Pathology, Faculty of Medicine,
University of Ljubljana, Ljubljana, Slovenia
3 Department of Ophthalmology, University Medical
Center Ljubljana, Ljubljana, Slovenia
Background and objectives: Of 350 million individuals
chronically infected with hepatitis B virus (HBV) worldwide,
approximately 15 million have been exposed to hepatitis D virus
(HDV) infection. As a result of vaccination against HBV, the preva-
lence of HDV has decreased in the last 20 years in the majority
of European countries, especially in Southern Europe. However,
it recently begun to rise again in some European countries, like
France, Germany and the United Kingdom, due to immigration from
endemic areas (mainly from Africa, Eastern Europe and Turkey).
Slovenia is a country with a population of approximately two mil-
lion and an estimated HBV prevalence of less than 5%. No reports on
HDV prevalence in Slovenia have been published to date in the peer
reviewed literature. The aim of our study was therefore to deter-
mine the HDV prevalence in Slovenian patients with chronic HBV
infection.
Materials and methods: Our study included 1305 HBsAg-
positive serum samples from the same number of patients
randomly selected from HBsAg-positive patients referred to the
Slovenian national reference laboratory for viral hepatitis between
February 1998 and December 2015. Considering the 95% confi-
dence interval and 2.5% margin of error, our sample size was
representative for all patients with chronic HBV infection in Slove-
nia. Serum samples were retrospectively tested for the presence
of total anti-HDV antibodies using commercially available ETI-AB-
DELTAK-2 (DiaSorin, Saluggia, Italy). Anti-HDV-positive samples
were further tested for the presence of anti-HDV IgM antibodies
and hepatitis D antigen (HDV-Ag) using commercially avail-
able ETI-DELTA-IGMK-2 (Diasorin) and ETI-DELTAK-2 (Diasorin),
respectively. Additionally, the in-house HDV reverse-transcription
real-time PCR, enabling amplification of a 71-bp fragment of
the conserved genomic region encoding HDV-Ag (J Clin Micro-
biol 2005;43:2363-9 ; J Clin Microbiol 2010;48:2022-9) and with the
analytical sensitivity of at least 300 viral copies/ml, was used to
determine the presence of HDV RNA in anti-HDV-positive samples.
Results: Total anti-HDV antibodies were detected in three out of
1,305 tested samples (0.23%; 95% confidence interval 0.08-0.67%).
Several consecutive serum samples were collected from all three
anti-HDV-positive patients. Apart from total anti-HDV antibodies,
no other HDV infection markers were detected in any of the tested
samples collected from a 48-year old male patient, indicating that
the patient has recovered from the past HDV infection. On the con-
trary, anti-HDV IgM antibodies and HDV RNA were detected in all
tested samples obtained from a 28-year old female patient and
a 60-year old male patient, suggesting the ongoing chronic HDV
infection.
Conclusions: In the first national prevalence study the observed
prevalence of HDV infection among HBsAg-positive patients in
Slovenia was surprisingly low, considering the fact that Italy,
with the HDV prevalence of 8.1%, is one of Slovenian neighboring
countries. Due to the observed low prevalence of HDV infection,
routine testing for HDV should not be considered in differential
diagnosis of exacerbation of liver disease in Slovenian patients with
chronic HBV infection.
Abstract no: 167
Presentation at ESCV 2016: Oral 29
Antiviral effect of interferons on BK virus
infection
E. Martin ∗, V. Descamps, V. Morel, F. Helle,
E. Brochot, G. Duverlie, S. Castelain, C. Franc¸ ois
Department of Virology, Amiens University Medical
Center and Virology Research Unit, EA4294, Jules
Verne University of Picardie, Amiens, France
The human polyomavirus BK (BKV) is a ubiquitous pathogen
that establishes an asymptomatic persistent infection in the
urinary tract of 80% of the human population. In immunocom-
promised patients, reactivation of the BKV infection is the cause
of nephropathy and hemorrhagic cystitis. Diseases associated
with BKV infections are increasing at the same time as potent
immunosuppressive therapies are developing. This highlights the
importance of components of the immune system in controlling
viral reactivation. However, the immune response to BKV, partic-
ularly the role of antiviral cytokines in infection control is poorly
documented.
Here, we investigated the antiviral effect of interferons (IFN)
on the BKV infection in renal cells. We tested IFN-alpha, lambda