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

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Abstracts / Journal of Clinical Virology 82S (2016) S1–S142 S21
Abstract no: 87
Presentation at ESCV 2016: Oral 38
Zika virus infections in travellers and contacts
in Lombardy, Northern Italy 2016
Francesca Rovida ∗, Giulia Campanini ,
Elena Percivalle , Maurizio Zavattoni ,
Antonella Sarasini , Alessia Arossa ,
Pierangelo Clerici , Paolo Antonio Grossi ,
Paolo Bonfanti , Fausto Baldanti
1 Molecular Virology Unit, Microbiology and Virology
Department, Fondazione IRCCS Policlinico San
Matteo, 27100 Pavia, Italy
2 Obstetrics and Gynecology Department, Fondazione
IRCCS Policlinico San Matteo, 27100 Pavia, Italy
3 Microbiology Unit, Hospital of Legnano, Milan, Italy
4 Department of Surgical and Morphological Sciences
of Clinical Medicine, Section of Infectious Diseases,
University of Insubria, Varese, Italy
5 Unit of Infectious Diseases, Manzoni Hospital,
Lecco, Italy
6 Department of Clinical, Surgical, Diagnostic and
Pediatric Sciences, University of Pavia, Pavia, Italy
Objectives: Zika virus infections in patients returning to Lom-
bardy Region (Northern Italy) and contacts were investigated.
Methods: serum samples of patients with potential Zika virus
(ZIKV) infections were tested for the presence of specific IgM and
IgG antibodies (Anti-Zika virus ELISA (IgM) and Anti-Zika virus
ELISA (IgG) by Euroimmun, Lübeck, Germany). Furthermore, the
presence of specific ZIKV antibodies was confirmed by plaque
reduction neutralization test (PRNT). Serum, saliva, urine and
semen samples, collected during the acute phase, were examined
for the presence of ZIKV RNAwith twomethods: a real-time reverse
transcriptase-polymerase chain reaction (RT-PCR) targeting a con-
served region of ZIKV and a pan-Flavivirus heminested RT-PCR
targeting a conserved region of the NS5 gene followed by sequenc-
ing of amplicons.
Results: in the period 18 February–20 April 2016, 5 confirmed
cases of Zika virus infection were diagnosed in Lombardy Region
(10 million inhabitants) in Northern Italy. Four (1 female and 3
male) patients had an history of recent travel, 1 arrived from the
Dominican Republic, 1 from El Salvador and 2 from Brazil, while
1 patient, the wife of the patient returning from the Dominican
Republic, had not travelled. Thus, a sexual transmission was doc-
umented. The patient returning from El Salvador was a 41 years
old pregnancy woman, 8 weeks gestation. During the symptomatic
phase of the infectionwere collected and analyzed for each patients
serum/plasma, saliva, urine and semen in males. Zika virus-RNA
was detected in 5/5 (100%) urine samples of the five patients, in
4/5 (80%) saliva samples while the viral genome was identified in
2/5 (40%) plasma samples. Furthermore, in 3/3 (100%) semen sam-
ples was detected Zika virus-RNA. In the symptomatic period Zika
virus specific IgM were detected in 3/5 (60%) patients while 0/5
(0%) specific IgG were detected. Zika virus-RNA has been detected
in serum up to 54 days, in urine up to 40 days, in saliva up to 11
days and in semen up to 19 days after onset of symptoms.
Conclusions: Zika virus infection shows a prolonged persistence
in peripheral blood with the potential of autochthonous spread to
competent mosquitoes. In addition, the presence of virus in semen
is an additional factor for autochthonous infections. Thus, sexual
partners of travelers must be included in surveillance protocols.
Abstract no: 81
Presentation at ESCV 2016: Oral 39
Zika virus infections imported to Portugal, the
National Reference Laboratory experience: The
importance of sample collection time lapse in
diagnosis
Líbia Maria Marques Zé-Zé∗, Maria João Alves
Centro de Estudos de Vectores e Doenc¸ as Infecciosas,
Instituto Nacional de Saúde Dr. Ricardo Jorge, Águas
de Moura, Portugal
Zika virus (ZIKV) belongs to the genus Flavivirus and was first
isolated from the blood of a febrile sentinel rhesus monkey dur-
ing a study of yellow fever in 1947, in Zika Forest, Uganda. ZIKV
is transmitted by Aedes sp. Mosquitoes, as Dengue, Yellow fever
and Chikungunya viruses, and was until recently considered a
mild pathogenic mosquito-borne flavivirus with very few reported
human cases of self-limiting acute febrile illnesses most often with
maculopapular rash, headache, malaise and conjunctivitis, all fol-
lowed by full recovery without sequel. During 60 years, cases of
ZIKV infections were detected only sporadically in Africa, and South
and Southeast Asia.
In 2007, the epidemiological history of ZIKV started to change
with a substantial outbreak in Yap Island (Federated States of
Micronesia), followed, since 2013, by outbreaks in French Poly-
nesia and other Pacific Islands. In French Polynesia a link relating
ZIKV infections with the increased incidence of Guillan-Barré syn-
drome and other neurological complications was assumed mainly
in regions with previous dengue epidemics. Nonetheless, at this
time, no onewas prepared for the dramatic outcomes of ZIKVarrival
in Brazil and, the explosive spread in the country and the huge
increase of several congenital malformations, microcephaly, and
other neurological disorders, including Guillain-Barré syndrome
and acute disseminated encephalomyelitis (ADEM) described since
2015. In February 1, 2016, the World Health Organization (WHO)
declared ZIKV infection a Public Health Emergency of International
Concern.
Genetic studies enable the identification of three distinct
genotypes: West African (Nigerian cluster), East African (MR766
prototype cluster) and Asian. All recent reported ZIKV outbreaks
have been associated to the Asian genotype. As most pathogenic
flavivirus, only a small percentage of ZIKV cases (estimated in ca.
25%) are symptomatic, and transmission via transfusion of infected
blood or organs donations, or sexual transmission, remains a risk.
The presence of other flaviviruses endemic in the same geographic
range is also a fact to take in consideration, as the incoming proves
that the eventual existence of antibodies to another flavivirusmight
facilitate a worsen development in ZIKV infection. On the other
hand, the clinical similarity of Zika and dengue virus infections and
the cross-reactivity of Zika antibodies with dengue viruses (DENV)
might have enabled the incorrect association of several Zika infec-
tions to DENV and certainly difficult serologic diagnosis.
Here, we discuss the clinical and laboratory aspects related to
some of the imported human cases of Zika virus in Portugal mainly
from Brazil, discuss the importance of time lapse in the choice of
sample and diagnostic analysis to achieve a confirmed ZIKV diag-
nosis result. Particularities of the diagnosis of secondary infections
by ZIKV after a probable primary DENV infection and the prob-
able ZIKV sexual transmitted infection in Madeira Island will be
presented.