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

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S56 Abstracts / Journal of Clinical Virology 82S (2016) S1–S142
Funding: Funding from the Hungarian National Development
Agency (TÁMOP-4.2.2.A-11/1/KONV-2012-0035).
Abstract no: 173
Presentation at ESCV 2016: Poster 70
RIDA® GENE Zika Virus: A new commercial
real-time RT-PCR assay for sensitive and
reliable detection of zika virus in urine and
serum samples
Lena Kastl ∗, Jessica Brückner
R-Biopharm A.G, Germany
Background: Besides endemic areas such as Africa and South
East Asia, high numbers of zika virus infections were recently
reported in South America, particularly in Brazil. In February 2016,
the World Health Organisation (WHO) issued a Public Health
Emergency of International Concern since microcephaly and other
neurological disorders were increasingly reported in newborns of
pregnant women with Zika virus infections. Zika virus belongs to
the genus of Flavivirus and similarly to other members of the Fla-
vivirus genus, transmission of zika virus occurs via mosquitos, in
particular mosquitos of the genus Aedes. Cross reactions with other
Flaviviruses, such as dengue virus or chickungunya virus, are often
observedupon antibody-specific diagnostic testing so that confirm-
atory testing is required, in particular in areas where there have
been possible co-infections.
Real-time PCR is a suitable method to specifically detect zika
virus RNA within the first week after onset of symptoms in serum
samples and in urine samples within 14 days after onset of symp-
toms.
This study aimed to evaluate a new real-time PCR assay for the
detection of zika virus in urine and serum samples.
Materials and methods: The RIDA® GENE Zika virusreal-time
RT-PCR assay detects zika virus-specific RNA by targeting the NS2A
gene. An internal control RNA detects PCR inhibition, monitors
reagent integrity and confirms successful nucleic acid extrac-
tion. The analytical reactivity and analytical specificity of the
RIDA®GENE Zika Virus was tested using known quality control
standards and reference materials. Spike experiments were carried
out to determine the analytical sensitivity. A clinical evaluation of
known positive and negative urine and serum samples was carried
out on the LightCycler® 480II (Roche) and compared to two other
commercially available test systems.
Results: Clinical evaluation of the RIDA®GENE Zika Virus with
known positive and known negative serum and urine samples
showed concurrent results when compared to two other com-
mercially available test systems. Commercially available zika virus
strains were used for evaluation of the analytical reactivity of
the RIDA® GENE Zika Virus assay. No cross-reactivity to other Fla-
viviruses including dengue virus, chickungunya virus and west nile
virus was detected with the assay. An analytical sensitivity of 50
copies/reaction was achieved with the LightCycler® 480II/LC2.0,
Mx3005P, Rotor-Gene Q, ABI7500, CFX96 and SmartCycler II real-
time PCR instruments.
Conclusions: The RIDA® GENE ZikaVirus real-time RT-PCR assay
is a sensitive and reliable assay for the detection of zika virus,
including both African and Asian variants. The assay is highly
specific for zika virus without known cross-reactivity to other fla-
viviruses such as dengue virus or chickungunya virus. The validation
of different common real-time PCR instruments provides broad
flexibility for use in the routine diagnostics laboratory.
Abstract no: 213
Presentation at ESCV 2016: Poster 71
Toscana virus meningitis in Southwestern
France
Camille Chagneau ∗, Jean-Michel Mansuy ,
Carole Barthe , Kévin Oliveira-Mendes ,
Jacques Izopet i Charrel
Catherine Mengelle
1 Department of Virology, Toulouse University
Hospital, Toulouse, France
2 Department of Physiopathology, Toulouse Purpan,
Unité Inserm U563, Toulouse, France
3 Department of Virology, Marseille University
Hospital, Marseille, France
Aims: Arboviruses are gaining more attention due to the
increased number of cases in human host. Toscana virus (TOSV) is a
member of Bunyaviridae andwas first identified in 1971 froma Phle-
botomus in central Italy. TOSV has a tropism for the central nervous
system and thus is recognized as an etiologic agent of meningitis
in the areas where it is present. The virus circulates in the Mediter-
ranean basin and during the warm seasons it might represent one
of the main causes of acute viral meningitis.
In France, the first cases were described among German travel-
lers returning from South East area in 1997 and the virus seems to
be present on a large Mediterranean coastal zone.
Also, in Southwestern France the vector Phlebotomus is circulat-
ing as evidenced by the presence of numerous canine Leishmaniosis
cases. But the incidence of aseptic meningitis linked to a TOSV
infection has never been studied in this area.
Material and methods: Patients suffering from aseptic menin-
gitis without any documented etiology were matched on sex and
age with patients suffering from documented enteroviral meningi-
tis (n = 58). All patients were attending, depending on age, children
or adults Toulouse University Hospital emergency unit.
We looked for Toscana virus, Herpes simplex virus 1 and 2
(HSV1-2) and Human herpes virus 6 (HHV6). Nucleic acids (NA)
were extracted from cerebro-spinal fluid (CSF) (samples stored at
−20 ◦ C pending batch analysis) with the MagNA Pure 96 DNA and
Viral NA Small Volume kit® on the MagNA Pure 96TM instrument
(Roche Molecular Diagnostics, Meylan, France) (input volume:
200 l, output volume: 100 l) according to the manufacturer’s
instructions.
Extracted NA were tested employing a monoplex in-house RT-
PCR for HSV1-2 and HHV6 on the Light Cycler (Roche Molecular
Diagnostics, Meylan, France). TOSV was tested using the CFX96TM
Real-Time System (Biorad diagnostics).
Results: We tested 66 CSF sampled from mid-april to mid-
october in 2014 and frommid-april to mid-october in 2015 among
patients (sex ratioM/F: 1.3) suffering from aseptic acute meningitis
negative for enterovirus. 21 patients were aged 5–10 year-old, 16
were 11–20, 16 were 21–30, 10 were 31–41 and 3 were older than
41 year-old.
No sample was positive for HSV1-2 and HHV6.
We detected however one sample positive for TOSV in a 36 year-
old woman’s CSF. She was suffering from a classical form of aseptic
meningitis (fever, headache and vomiting) associated to myalgia
but without photophobia and was discharged after three days of
hospitalization with favorable outcome.