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

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Abstracts / Journal of Clinical Virology 82S (2016) S1–S142 S69
Abstract no: 177
Presentation at ESCV 2016: Poster 96
Characterization of NS3 region and frequency of
resistance mutations against Simeprevir of
hepatitis C virus genotype 1a and 1b in
Portuguese infected patients
R. Marcelino ∗, J. Cabanas , M.F. Goncalves ,
A.P. Carvalho , I. Costa , I. Diogo ,
J.M. Marcelino , P. Gomes
1 Global Health and Tropical Medicine (GHTM), Unit
of Teaching and Research of Medical Microbiology,
Instituto de Higiene e Medicina Tropical (IHMT),
Universidade NOVA de Lisboa (UNL), Portugal
2 Molecular Biology Lab. LMCBM, SPC, HEM – Centro
Hospitalar Lisboa Ocidental, Lisboa, Portugal
3 Centro de Investigac¸ ão Interdisciplinar Egas Moniz,
CiiEM, ISCSEM, Almada, Portugal
Hepatitis C virus (HCV) is a significant public health problemand
the leading cause of liver transplantation and hepatocellular carci-
noma. Globally, approximately 180million people are infectedwith
HCV. There are seven genotypes of HCV, with a variety of subtypes.
The Portuguese prevalence of HCV infection is about 1.0%, being the
genotype 1 the most prevalent and one of the most difficult to treat
with the previous therapies. However, the face of HCV therapy has
changed dramatically since 2011 with all-oral regimens that have
been emerging, characterized by high genetic barrier to viral resis-
tance and lowside effects. Anewhope to cureHCV infected patients
have arisen. Actually, in Portugal the first line treatment is Sofos-
buvir plus Ledipasvir and in fact we have around 95% of sustained
viral response from the treated patients. Those who still do not cure
infection are proposed to Sofosbuvir plus Simeprevir, being the last
one ineffective if in the genome of the virus are present somemuta-
tion such as Q80K, S122R/A/G, R155K/QD168E/V/H/A and I/V170T.
Thus, the characterization of NS3 mutations has a highly impact on
the prediction of the success of this direct antiviral agent. Q80K is
observed predominantly in HCV genotype 1a as a naturally occur-
ring polymorphism and seldom in other HCV genotypes. It ranges
from 5% to 47%, depending on geographic region.
The objective of this study was to characterize HCV NS3 of Por-
tuguese patients still naive to Simeprevir, once less is known in our
population about the occurrence of natural mutations to this drug
in that viral genomic region.
HCV NS3 region of 112 patients infected with 1a (89.3%) and 1b
(10.7%) viral genotypeswas sequencedwith an in house sequencing
method described previously by Harringan et al. and the analysis
of NS3 amino acids 1–181 was performed using geno2pheno HCV.
The prevalence of known pre-existing resistancemutationQ80K
was 3.36% in the whole sample. However, it was not found in geno-
type 1b and this turned the prevalence of Q80K in genotype 1a
around 3,0%. Q80R resistance mutation was found in one sample of
genotype 1a (1%) as other pre-existing polymorphisms at that same
position, Q80H (2%), Q80L (1%) which are not associated to resis-
tance but may act as intermediate mutations to Q80K. Additionally,
it was found R155K (1%) and R155G (1%) in genotype 1a, both con-
ferring resistance to Simeprevir. Notably, in genotype 1a there was
also a higher occurrence of the substitutions T40A (62%), N174S
and N174G (71%), S91A (98%) and L153I (100%) which have rarely
been reported in the literature. T40A and S91A involve changes in
the amino acid’s electrical charge and position 91 is in close prox-
imity to the residues in the catalytic triad. L153I does not lead to
this kind of amino acid’s changes, but based on its high frequency
in this population, it may be a regional genetic polymorphism and
surprisingly, there were found 71% of samples simultaneously pre-
senting L153I, S91A andN174S/Gwhichmay suggest an association
of substitutions.
In conclusion, there were not just detected 5% of preexisting
mutations that cause resistance to Simeprevir, but also some poly-
morphisms that should be further investigated not only by their
notably association, but also because the changes they promote
near the catalytic center of the enzyme, that can be a cause of some
still unknown protease inhibitor treatment failures too.
Abstract no: 178
Presentation at ESCV 2016: Poster 97
Hepatitis B virus infection and reactivation in
two patients with anti-HBs following
etanercept therapy and alemtuzumab and
rituximab therapies: Case presentation
Selda Erensoy ∗, Emine Emel Koc¸ man ,
Rüc¸ han Yazan Sertöz , Vedat ˙Inal , Zeki Karasu
1 Ege University Faculty of Medicine Department of
Medical Microbiology, Turkey
2 Bayburt State Hospital Medical Microbiology
Laboratory, Turkey
HBV reactivation is a well known phenomenon following cyto-
toxic or other immunosuppressive anticancer therapy. Biologic
therapies with monoclonal antibodies lead to hepatitis B virus
(HBV) reactivation. It is aimed to present and discuss two hep-
atitis B cases following therapies with monoclonal antibodies to
emphasize HBV assessment in patients receiving biologic therapy.
Materials and methods: HBV serological test results are eval-
uated with the previous markers of the cases. HBsAg, anti-HBc,
IgM anti-HBc and anti-HBs are tested with Architect system
(Abbott Diagnostics, Germany). HBV viral load is testedwith Abbott
m2000sp and m2000rt (Abbott Molecular Diagnostics, USA). Cases
with markers of HBV infection who were HBsAg negative and anti-
HBs positive are evaluated retrospectively andwith clinical history.
Case 1: A man with rheumatoid arthritis had HBV DNA 7.14
log10IU/ml, HBsAg positive, anti-HBc positive, anti-HBe positive,
anti-HBs negative, IgM anti-HBc negative. Liver function tests
were elevated (>2N). He received lamivudin and seroconverted
to anti-HBs positivity, HBsAg negative, but HBV DNA was still
1.34 log10IU/ml. Liver function tests turned to normal values. He
was HBsAg negative, anti-HBc positive and anti-HBs 13mIU/ml six
months ago. He had received etanercept therapy six months ago.
It was found that he had chronic HBV infection four years ago, but
seroconverted in three years. He had no HBV DNA data beforehand,
but found to be positive as 2.18 log10IU/ml when tested in archived
samples.
Case 2: 64 years old male with cirrhosis was found to be HBsAg
and HBV DNA positive (8.44 log10IU/ml), anti-HBc positive, anti-
HBs negative. He was HBsAg negative, anti-HBc positive, anti-HBs
232mIU/ml before. He had chronic lymphocytic leukaemia-small
lymphocytic lymphoma and received alemtuzumab four years ago.
He had graft versus host disease after allogeneic bone marrow
transplantation and received rituximab three years ago. At that
time he was HBsAg and anti-HBc negative, anti-HBs 159mIU/ml.
Conclusion: Hepatitis may be observed also in subjects with
anti-HBs positivity or with occult HBV infection with markers of
previous exposure to HBV. HBV assessment in patients receiving
biologic therapy (anti-TNF-alpha, anti-CD20, anti-CD52) is impor-
tant and should be always remembered. HBsAg, anti-HBc, anti-HBs,
ALT and AST should be tested. Quantitative HBV DNA test should
be applied if HBsAg and/or anti-HBc are positive. Antiviral ther-