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

110 / 152

S106 Abstracts / Journal of Clinical Virology 82S (2016) S1–S142
dried and calibrated where possible to the appropriate Interna-
tional Standard.
Second, NIBSC have developed a web based Results Reporting
System (RRS) to data monitor both serology and PCR reagents. The
system allows for real-time comparison of both intra- and inter-
laboratory results by provision of Levey–Jennings plots and indi-
cates when an assay is falling out of specification. By application of
Westgard Rules to the data, any deviations from the norm can be
quickly recognised and addressed.
Medical Diagnostic Laboratories who are audited against ISO
15189 are required to use “suitable reference material” and where
samples are tested at different laboratory sites there should be a
mechanism to “verify the comparability of results”. Use of the ref-
erence materials provided by NIBSC together with the RRS allow
a medical diagnostic laboratory to meet the requirements of ISO
15189.
Abstract no: 206
Presentation at ESCV 2016: Poster 172
Evolutionary studies of herpes simplex viruses
(HSV) genomes provide evidences of
HSV-2/HSV-1 interspecies recombination
S. Burrel ∗ . Boutolleau Ryu Agut
K. Merkel , F. Leendertz , S. Calvignac-Spencer
1 Sorbonne Universités, UPMC Univ Paris 06, CR7,
CIMI, INSERM U1135 and AP-HP, Hôpitaux
Universitaires Pitié-Salpêtrière – Charles Foix,
Service de Virologie, Paris, France
2 Robert Koch Institute, Epidemiology of Highly
Pathogenic Microorganisms Unit, Berlin, Germany
Herpes simplex virus 2 (HSV-2) is a prevalent sexually transmit-
ted infection responsible for recurrent genital lesions and may also
cause neonatal morbidity and mortality. The average prevalence is
around 11% but important regional variations exist, with the high-
est prevalence observed in sub-Saharan Africa (31.5%) HSV-2
generally exhibits low genomic variability. The maximum overall
divergence is only 0.4% andmost open reading frames (ORF) exhibit
little, if any, variability ecently, we described an HSV-2 vari-
ant mainly found in sub-Saharan African individuals characterized
by highly divergence among UL30 gene (maximum divergence of
2.4%) n order to clarify the evolutionary history of this variant,
sequences of nearly complete genomes were obtained from 18 iso-
lates of HSV-2 variant recovered from distinct patients originating
from Africa.
Sequencing libraries were prepared using extracted DNA from
supernatants of infected cell cultures, then were subjected to
in-solution hybridization capture and sequenced on a MiSeq®
platform (Illumina) with a resulting average coverage of the
genome sequences about 75%. Whole genome sequence com-
parisons revealed unexpected diversity, with many sequences
exhibiting more than 0.7% pairwise divergence. Phylogenetic anal-
yses identified two main lineages: a previously unrecognized
African lineage, mostly comprising sequences originating fromsub-
SaharanAfrica, and aworldwide-spread lineage, even distributed in
sub-Saharan Africa. Recombination analyses performed thereafter
notably evidenced that members of both lineages could recom-
bine among themselves. Moreover, those analyses also showed that
interspecific recombination might have occurred between HSV-2
and HSV-1 ORF fragments, as evidenced for UL29 and UL30, and,
to a lesser extent, for UL15 and UL39. The recombination status at
these loci was used to investigate the relative timing of the recom-
bination events. While the recombination events in UL15 and UL39
appeared after the African and worldwide-spread lineages started
their diversification, the recombination events in UL29 and UL30
likely constituted the milestone of the worldwide-spread lineage
origin.
Those results highlight the potential African origin of HSV-2,
which is coherent with human species evolutionary history, and
assess, unlike to common belief, the occurrence of interspecies
HSV-2/HSV-1 recombination under natural conditions The
extended gene flow from HSV-1 into HSV-2 genomes may have
contributed to the rise of the worldwide-spread lineage. Further
investigations are now required in order to determinewhether HSV
interspecific recombination is still an ongoing process and has any
clinical implications.
Reference
[1] K.J. Looker, A.S. Magaret, K.M. Turner, P. Vickerman, S.L. Gottlieb, L.M. Newman,
Global estimates of prevalent and incident herpes simplex virus type 2
infections in 2012, PLOS ONE 10 (2015) e114989.
[2] R.M. Newman, S.L. Lamers, B. Weiner, et al., Genome sequencing and analysis
of geographically diverse clinical isolates of herpes simplex virus 2, J. Virol. 89
(2015) 8219–8232.
[3] S. Burrel, N. Desire, J. Marlet, et al., Genetic diversity within
alphaherpesviruses: characterization of a novel variant of herpes simplex virus
2, J. Virol. 89 (2015) 12273–12283.
[4] E. Thiry, F. Meurens, B. Muylkens, et al., Recombination in alphaherpesviruses,
Rev. Med. Virol. 15 (2) (2005) 89–103.
Abstract no: 242
Presentation at ESCV 2016: Poster 173
Commutability and agreement of international
and secondary standards
M.K. van Genne∗, R. de Boer, R. Poelman,
H.G.M. Niesters
University Medical Centre Groningen, The
Netherlands
Background: Quantitative testing of viral loads has become
an integral part of care for immunocompromised patients. Infor-
mation on the commutability of quantitative assays is currently
not available due to differences in methodology, chemistry and
equipment. The WHO international standards and secondary stan-
dards have been developed for only a few viruses associated with
these immunocompromised patients, aiming to improve informa-
tion on commutability and to standardise diagnostics between
laboratories. This study aims to investigate the commutability of
international standards, as well as to evaluate several secondary
standards.
Material/methods: WHO international standards as well as
commercially available secondary standards were compared using
real-time PCR as well as digital PCR technology. We investigated
the commutability of international standards and the agree-
ment between several secondary standards for the same targets.
Cytomegalovirus, Epstein–Barr virus, BK virus, Varicella Zoster
Virus, Hepatitis A virus, Herpes Simplex virus type 1 and 2 were
included.
Results: The international as well as secondary standards
indicated a high correlation and suitability to improve diag-
nostics. WHO international standards for Cytomegalovirus and
Epstein–Barr virus showed significantly. However, when no inter-
national standard is present for a target, the agreement between
secondary standards was significantly lower. The data collected