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

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S90 Abstracts / Journal of Clinical Virology 82S (2016) S1–S142
Quality trimming and data analysis were performed with the AVA
software.
Results: LMP1 genewas successfully amplified and sequenced in
131 samples from88 patients (i.e. LMP1 was detected in 35 (63.6%),
24 (51.1%) and 3 (21.4%) of the WB and 23 (51.1%), 35 (74.5%) and 6
(42.9%) of the OC for HIV+/HL+, HIV+/HL − and HIV−/HL+ patients,
respectively. The LMP1 gene from HIV+/HL+ patients could only be
amplified in 5 (26.3%) out of 19 tumor biopsies.
The frequency of WT-LMP1 and del30-LMP1 was not different
between the 3 groups of patients regardless the sample analysed.
The Del69-LMP1 variant was only detected in HIV+ patients. In
21.4% of cases (24 patients), the variant detected in the patient’s
WB and OC specimens was different.
In 4 out of 5 biopsies of HIV+/HL+ patients, only one LMP1 type
was detected (3 WT-LMP1 and 1 del30-LMP1). In the fifth biopsy
the del30-LMP1 variant was dominant, represented at 99% with
only 1% ofWT-LMP1. For only two of these patients, we successfully
sequenced LMP1 in all biological compartments (WB, OC and tumor
biopsy). One patient presented the same del30-LMP1 variant in the
3 compartments. The second patient harbored only the WT-LMP1
in the tumor biopsy and both WT-LMP1 and del30-LMP1 in the 2
other compartments (60/40% and 50/50%WT-LMP1/del30-LMP1 in
WB and OC, respectively).
In 17 out of 131 samples (13%), the NGS technology allowed
to detect LMP1 variants with a sensitivity below 15% and as low
as 0.5%. Furthermore, in the sequenced LMP1-region, 85 sporadic
mutations were observed, 74 of which were non-silent. Among
these substitutions, 24 are located in the CTAR2 domain of LMP1.
Conclusion: Further analysis of tumor biopsy are needed (work
in progress) but this preliminary study showed the heterogeneity of
the LMP1wild type or LMP1 variants detectedwith NGS technology
without clear differences in the frequency of LMP1wild type versus
LMP1 variant among HL+ or HL− patients.
Abstract no: 236
Presentation at ESCV 2016: Poster 139
Assessment of laboratory performance in the
molecular detection of HIV through
International EQA Scheme
N. Kivi ∗, E. Tuovinen , M. Lappalainen
1 Labquality Oy, Helsinki, Finland
2 Department of Virology and Immunology, Helsinki
University Hospital Laboratory Services (HUSLAB),
Helsinki, Finland
Various nucleic acid assays have been developed for diagnos-
tics and therapeutic monitoring of acute human immunodeficiency
virus (HIV) infection. Early detection of these acute viral infections
is vital and considering that these viruses have high replication
rates, early detection upon transmission is crucial. Viral monitor-
ing and early viral load may allow the infection to be halted before
clinical symptoms are apparent. Additionally changes in viral load
in the patient may indicate the need to modify treatment strategies
to prevent further disease progression.
The generation of reliable and reproducible test results is there-
fore of great importance in clinical settings. The development of
international standards has helped considerably in improving test
reproducibility within the laboratory, which in turn helps facil-
itate comparison of results across laboratories. External Quality
Assessment (EQA) schemes provide a valuable tool in support of
standardization. The design of these schemes allows comparison
across multiple laboratories using different molecular assays.
We report the first year’s results of recently launched interna-
tional EQA scheme focused on the nucleic acid detection of HIV.
Altogether 22 laboratories from9 different European countries par-
ticipated in five HIV scheme rounds organized during 2015-2016.
Eleven different commercial assays were reported and most com-
mon methods were RealTime HIV-1 assay (Abbott) and COBAS
Ampliprep/COBAS Taqman HIV-1 assay (Roche). Method used for
pre-testing value determination performed by expert laboratory
was Cobas Ampliprep/COBAS TaqMan HIV-1 v2.0 (Roche) in each
round. Reported results were statistically evaluated and showed
a trend of excellent qualitative performance. All assays used in
laboratories indicated a high degree of specificity. EQA scheme
is suitable for qualitative and quantitative analyses. Results are
scored based on qualitative results, but laboratories report also
quantitative results, when applicable, and many laboratories also
reported copy number results. Overall precision in quantitative
determination has been fairly good. Interestingly, in quantitative
results notable differences compared to the pretested values were
observed in every round. As high as ten-fold difference in copy
number count were observed between participants and as com-
pared to the pretesting result. The results from this first year of the
scheme strengthen the importance of external quality assurance
programs for HIV nucleic acid quantification to ensure the quality
of testing and diagnostics.
Abstract no: 244
Presentation at ESCV 2016: Poster 140
Analysis of the mRNA expression of DNA
damage response genes in HIV infected patients
D. Di Carlo ∗, A. Fantauzzi , T. Melengu ,
P. Maida , S. Serafino , N. Giustini , S. Tozzi ,
G. Antonelli , F. Falasca , O. Turriziani
1 Department of Molecular Medicine, Sapienza
University of Rome, Italy
2 Department of Clinical Medicine, Sapienza
University of Rome, Italy
3 Department of Public Health and Infectious
Diseases, Sapienza University of Rome, Italy
Though combined antiretroviral therapy (cART) in HIV-1 pos-
itive patients allows a massive suppression of viral replication,
many aspects of persistence and pathogenesis of infection are still
unknown. It has been reported that virus-induced cell killing is trig-
gered by viral integration. Infection by wild-type HIV-1, but not
an integrase-deficient mutant, induced the death of activated pri-
mary CD4 lymphocytes. Similarly, integrase inhibitors abolished
HIV-1-induced cell killing both in cell culture and in CD4 + T cells
from acutely infected subjects. The mechanism of killing during
viral integration involved the activation of DNA-dependent protein
kinase (DNA-PK), a central integrator of the DNA damage response,
which caused phosphorylation of p53 and histone H2AX.
The aim of the study was to evaluate if Inhibitor Integrase (INI)
containing regimen could affect mRNA expression profile of DNA
damage response genes involved, in HIV infected patients.
Forty PBMC samples from HIV+ patients (18 treatment naïve
and 22 treated with ART containing INI) and 10 sample from
healthy donors (HD) were collected; mRNA levels of FasR, XRCC1,
Lig III , Parp-1, DNA PkI, DNA PkII were evaluate using Syber
GreenReal time PCR (Agilent Technologies). All HIV treated patients
had undetectable viremia. Results were normalized using house-
keeping genes beta-actin ( CT). The fold-difference of expression
levels between three groups were measured comparing CT val-