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

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Abstracts / Journal of Clinical Virology 82S (2016) S1–S142 S45
comparatively short turnaround time of two days the workflow
offers relevant improvements in HIV DRM detection.
Abstract no: 351
Presentation at ESCV 2016: Poster 48
Mass spectrometry of influenza virus using
clinically available MALDI-TOF platform
Andrei Musaji ∗, Richard Fahlman,
Carmen Charlton
University of Alberta, Canada
Viral respiratory disease caused by influenza viruses has an
importantmedical, epidemiological and economic impact on global
population. Ideal screening assays for influenza viruses directly
from clinical specimens must be not only of high sensitivity and
specify, but also must have short turn-around times (less than
two hours). In many laboratories, screening for influenza virus
directly from clinical specimens is based on express Direct Fluores-
cent Antibody Assay, which is quite specific, but not very sensitive,
when compared to RT-PCR (reverse transcriptase PCR). On the other
hand, nucleic acid amplification assays, such as RT-PCR, are not
characterized by quick turn-around times. Little is known if mass
spectrometry technology may be used as an alternative screening
approach for influenza virus identification directly from clinical
samples.
The main objective of our study was to analyse mass spectra
for influenza virus identification using a mass spectrometer, which
is routinely used in our Clinical Microbiology Diagnostic Labora-
tory for bacterial or fungal identification. In this preliminary study,
we used cell culture or egg-amplified influenza viruses as well as
their corresponding recombinant neuraminidase (NA) and hemag-
glutinin (HA) proteins. Mass spectrawere generated using clinically
available Biomerieux Vitek® MS MALDI-TOF mass spectrometer.
All proteins and whole viruses were pre-treated by lysis solution,
sonication, boiling and microwaving. Influenza viruses and their
corresponding HA and NA proteins were either directly applied on
MALDI-TOF target or a minimal pre-treatment with either formic
acid extraction and/or short trypsin digestion was used prior to
target application of samples. Overall, the turn-around time for
specimens was from less than an hour (without trypsin digestion)
to three hours (with trypsin digestion). Mass spectra for H1N1,
H3N2 and B influenza viruses as well as their corresponding HA
and NA were analysed. We used VENN diagrams to manually ana-
lyze spectra of HA and NA and the corresponding whole virus in
order to identify potential peak (m/z) candidates for the influenza
virus identification by MS MALDI-TOF. On average, three to five
peak candidates were identified for each influenza virus based on
mass spectra analysis of HA, NA and entire influenza virus mass
spectrometry.
Engineering and implementation costs for a mass spectrome-
ter, which may be used in a Clinical Microbiology setting and yet
possess a resolution capacity comparable to that of research use
only platforms, is quite lengthy and financially demanding proce-
dure. In this preliminary study we investigated whether already
clinically available mass spectrometer may be used for influenza
virus identification. We obtained a number of peak candidates,
whichmay be used for a peak database creation in the future exper-
iments. In the future, we are planning to identify influenza viruses
directly from virus-spiked clinical specimens as well as from clini-
cal specimens derived from symptomatic patients.
Abstract no: 41
Presentation at ESCV 2016: Poster 49
Increasing blood safety by diagnosing Zika,
Chikungunya and Dengue in times of massive
outbreaks
A. Latz , M.E. Pereira , R. Berlie
1 NovaTec Immundiagnostica GmbH, Dietzenbach,
Germany
2 Siemens Healthcare GmbH, Marburg, Germany
3 NovaTec Immundiagnostica GmbH, Germany
The Aedes trio (Dengue, Chikungunya and Zika) are arthropod-
borne viruses that are transmitted by mosquitos of different Aedes
species (Aedes aegypti, Aedes albopictus). They have been reported
in Africa, the Americas, Asia and the Pacific Islands. Dengue virus is
a flavivirus, closely related to Zikawhereas Chikungunya belongs to
the alphaviruses. Dengue shares some clinical signs with Chikun-
gunya and Zika and they can be misdiagnosed in areas where these
arboviruses are common. As Dengue infectionmay cause a rash that
could be confused with other diseases such as Chikungunya, Zika
and measles, these other diseases do need to be ruled out. Diagno-
sis of Zika will first and foremost be by exclusion of other diseases
such as Chikungunya and Dengue, based on symptoms and travel
history. It is known that these diseases can also be transmitted by
blood transfusion. Since a great proportion of infected persons are
asymptomatic special care has to be taken in respect to blood safety.
Surveillance and testing algorithm for these three co-circulating
arboviruses are needed since they show high impact on the
socio economic burden in endemic countries. WHO proposed very
recently a testing guidance for laboratory detection and diagnosis
of these diseases. On 1 February 2016 the WHO declared a Pub-
lic Health Emergency of International Concern (PHEIC) regarding a
recent cluster of microcephaly cases and other neurological dis-
orders and the possible association of these illnesses with Zika
virus infections. TheWHO recommended efforts towards improved
surveillance of Zika virus which is only possible with an accurate
diagnostic system for Dengue and Chikungunya as well.
Here we showhow the BEP® III and BEP 2000 Advance ® systems
of Siemens Healthcare GmbH in combination with the Novagnost®
ELISA assays can help in the management of outbreaks, proper
diagnosis of individuals and surveillance of populations at risk.
The combination of highly sensitive and specific ELISA
assays (Dengue IgG sensitivity >95%/specificity >95%; Dengue
IgM sensitivity 82,3%/specificity >95%; Chikungunya IgG sen-
sitivity >98.6%/specificity 100%; Chikungunya IgM sensitivity
>98.8%/specificity 100%/Zika IgM sensitivity >100%/specificity
98.2%) fulfills the criteria of the WHO testing guidance for high
throughput screening of these diseases and therefore seems to be
an excellent tool for surveillance of blood products.