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

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S88 Abstracts / Journal of Clinical Virology 82S (2016) S1–S142
Conclusions: HIV-1 molecular epidemiology studies in the
HBV ± HDV and HCV co-infected patients are important tools for
tracking transmission patterns and the spread of CRF and monitor-
ing of CRF subtypes of HIV-1 in globally scale may be important
in vaccine development against HIV. However, the high prevalence
of HIV-1 ART resistance mutations in such as patients suggested
that the resistance testing must be an integral part of the manage-
ment of HIV-1 infection and the choice of first – line therapy regime
should be guided by the results of genotypic resistance in Turkey
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Abstract no: 20
Presentation at ESCV 2016: Poster 135
Genotyping of HPV DNA positive and HPV E6/E7
mRNA negative cervical samples with abnormal
cytology
A. Altay ∗, I. Tuney , K. Ergunay , A. Usubutun ,
K. Yuce , A. Pinar , I. Gorzer ,
E. Puchhammer-Stockl , G. Bozdayi
1 Gazi University, Faculty of Medicine, Department of
Medical Microbiology, Ankara, Turkey
2 Hacettepe University, Faculty of Medicine,
Department of Medical Microbiology, Virology Unit,
Ankara, Turkey
3 Hacettepe University, Faculty of Medicine,
Department of Pathology, Ankara, Turkey
4 Hacettepe University, Faculty of Medicine,
Department of Obstetrics and Gynecology, Ankara,
Turkey
5 Medical University Vienna, Department of Virology,
Vienna, Austria
Background: Human papillomaviruses have been established
as a risk factor for invasive carcinoma of the uterine cervix. HPV
DNA detection, provides an efficient method of screening. Detec-
tion of the HPV E6/E7 oncogene expression emerged as a promising
biomarker to determine the risk for the progression to high-grade
cervical lesions. In the present study, we aimed to determine the
genotypes of HPV DNA positive and mRNA negative samples from
our previous study.
Material and methods: HPV mRNA and DNA detection in
samples with abnormal cytology were evaluated in our previous
study. Cervical specimens were obtained at Hacettepe University
Hospital, Department of Obstetrics and Gynaecology via cervi-
cal brushes during January–October 2011. Real-time PCR (Heliosis
Human Papilloma Virus LC PCR Kit, Metis Biotechnology, Turkey)
and NASBA assay (NucliSENS EasyQ HPV v1.1, bioMerieux, France)
were performed to detect HPV DNA and E6/E7 mRNA, respec-
tively. In house PCR method was used for genotyping of HPV
type 18, 31, 33, 35, 39, 45 and detection of E6/7 gene with
PCR master mix (Promega Corporation, Madison, WI, USA), PGMY
(Metis Biotechnology, Turkey) and E6/E7 consensus primers (Heliks
Biotechnology, Turkey). The primer sequences were controlled in
BioEdit 3.0 programme comparingwithHPV reference strains. Con-
firmation of the samples was done by ‘Line Probe Assay’ (INNO-LiPA
HPV Genotyping Extra Amp, Innogenetics, Belgium).
Results: Totally 81 women with abnormal cytology result con-
stituted the previous study group. HPV DNA was identified in 73
samples (90.1%) that comprise HPV-16 in 46 samples (63.1%), HPV
other than 16 in 15 samples (20.5%) andmixed HPV infections in 12
samples (16.4%). HPV E6/E7 mRNA expression was observed in 45
samples (55.6%). Towards these results, we tried to determine the
genotypes of 28 HPVmRNA negative-type 16 DNA positive samples
and 15 HPV DNA other than type 16 positive samples; totally 43
samples were tested. Among the 43 samples, 5(12%) samples were
detected positive with E6/E7 consensus primers. Four of themwere
HPV-16 and 1 was HPV genotype other than 16. PCR with positive
controls which belongs to genotypes 18, 31, 33, 39 and 45 were
carried out. All positive controls worked except HPV-33. There-
fore, a new primer for HPV-33 (TIB MOLBIOL, Berlin, Germany)
was designed and used. However, it did not work again, although
it was tested in several PCR conditions. Beside this, all 43 samples
were negative for genotypes 18, 31, 35, 39 and 45. Therefore, con-
firmation of the samples was further done by using LiPA. Totally 12
samples were tested by LiPA. Genotypes detected by LiPA were in 8
cases different from that investigated by PCR (6, 11, 16, 43, 53, 62,
81, 89). Two samples were identified as HPV-33 and two others as
HPV-39 by LIPA. However, in none of these 4 samples, these HPV
genotypes could be detected by PCR. Because of the limited amount
of sample material we could not test all samples by LiPA.
Conclusion: As a result, depending on LiPA findings, it seems
that a number of samples investigated were detected negative by
PCR because they had genotypes other than genotypes 18, 31, 35, 39
and 45. Beside this, LiPA amplifies a 65-bp region, and its sensitivity
thus might be higher than our PCR method which amplifies 238-
bp and 455-bp long regions. Also the E6/E7 gene region is not as
conserved as the L1 gene region therefore it is possible to obtain
more negative results with E6/E7 PCR.
Abstract no: 205
Presentation at ESCV 2016: Poster 136
Performance of the LIAISON ® XL murex
recHTLV-I/II assay
T.D. Ly∗, C. Coignard
Laboratoire Biomnis, France
Background: There are two types of HTLV: HTLV-I and HTLV-II.
HTLV-I is associated with adult T-cell lymphoblastic leukemia and
B-cell chronic lymphocytic leukemia and a demyelinating disease
called HTLV-I associated myelopathy/Tropical spastic paraparesis
(HAM/TSP). It is estimated that 15–20 million people are currently
infected with human T-cell lymphotropic virus type 1 (HTLV-1)
worldwide. HTLV-II is less common and is associated with neo-
plasias of the CD8T lymphocytes. Transmission of both HTLV I and
II occurs through sexual contact, exposure to blood, transfusion of
infected cellular blood components and perinatally, probably by
breast feeding.
Aim: In this study, the performance of LIAISON® XL murex
recHTLV-I/II assay (DiaSorin, Saluggia, Italy) was compared to that
of ARCHITECT rHTLV-I/II (Abbott, Wiesbaden, Germany), used rou-
tinely in our laboratory.
Methods: During a 2 week period, all unselected serum sam-
ples (N = 663) submitted to the laboratory for HTLV testing were
examined by LIAISON® XL murex recHTLV-I/II assay. Samples that
were discordant were tested by INNO-LIA HTLV I/II Score (Fujirebio