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

62 / 152

S58 Abstracts / Journal of Clinical Virology 82S (2016) S1–S142
Parvovirus B19, a well-characterized human pathogen can rarely
cause encephalitis, our findings did not confirm such an associa-
tion for BuV in this preliminary investigation. However, long-term
evaluation of individual cases with unknown etiology is required
and might reveal this virus to involved in certain settings.
Abstract no: 330
Presentation at ESCV 2016: Poster 74
Mumps outbreak among vaccinated students in
Trondheim, Norway in 2015
S.A. Nordbø ∗, S. Krokstad , A. Christensen ,
K.S. Borge agvik
1 Department of Medical Microbiology, St. Olav’s
Hospital and Department of Laboratory Medicine,
Children’s and Women’s Health, Faculty of Medicine,
Norwegian University of Science and Technology,
Trondheim, Norway
2 Department of Medical Microbiology, St. Olav’s
Hospital, Trondheim, Norway
3 Department of Virology, Norwegian Institute of
Public Health, Oslo, Norway
4 City of Trondheim, Department of Infectious
Disease Control, Municipality of Trondheim, Norway
Background: Mumps vaccination (genotype A) was introduced
in the Norwegian childhood vaccination schedule in 1983 with
vaccination coverage exceeding 90%. At the end of August 2015,
a foreign student was hospitalized at St. Olavs Hospital in Trond-
heim with parotitis and orchitis, and during the period September
2015 to January 2016, 176 suspected cases were registered within
the municipality of Trondheim.
Material and methods: Diagnosis was confirmed by an in-
house PCR, viral culture and/or serology. Specimens from the oral
cavity was collected by flocked swabs in UTM-medium (Copan),
and was the method of choice. Isolates were sent to Norwegian
Institute of Public Health in Oslo for genotyping.
Results: Of 148 confirmed cases, 127 were students, and the
vast majority had been vaccinated against mumps. Seven patients
were hospitalized, six had orchitis and one patient had meningitis.
The health authorities in Trondheim vaccinated close contacts and
unvaccinated students. 53 isolates were genotyped, and they were
all genotype G. Mumps PCR was positive until 11 days after onset
of symptoms. Furthermore, the virus could be cultured until 9 days
after symptom debut. The sensitivity of nasopharyngeal and urine
specimens was too low to be used for diagnostic purposes. EBV-
DNA was detected in 13 of 27 (47%) specimens testing positive for
mumps virus RNA, but in low concentration. Viral culture proved
to be important for confirmation of the first cases, and to adjust the
sensitivity of the in-house PCR. Only one (5%) out of 20 vaccinated
students with confirmed mumps infection had detectable serum
IgM using the LIAISON® Mumps IgM assay.
Conclusion: The outbreak of mumps among vaccinated stu-
dents suggests that the currentmump vaccinesmay not be effective
in preventing genotype G mumps outbreaks. Serological methods
often fail to detect mumps infections in immunized patients, and
PCR from specimens taken from the oral cavity is the test of choice
to diagnose these infections.
Abstract no: 334
Presentation at ESCV 2016: Poster 75
Analytical performance and method
comparison of the VERSANT Zika RNA 1.0 Assay
(kPCR)
T. Battersby ∗, A. Chmura, S. Hao, H. Huang,
G. Kritikos, A. Lal, R. Malhotra, A. Miller,
D. Monga, J. Mosner, A. Patel, C. Wagner
Siemens Molecular Diagnostics, United States
Background: Zika virus (ZIKV) is a mosquito-borne virus of
the family Flaviviridae first isolated in 1947 in Uganda. The first
ZIKV outbreak outside of Africa and Asia occurred in 2007 in Yap
Island (Federated States of Micronesia). The largest outbreak was
from October 2013 to March 2014 in French Polynesia (FP), Pacific.
The World Health organization (WHO) recently declared that the
ongoing recent cluster of microcephaly cases and other neurologi-
cal disorders reported in the Americas, constitutes a Public Health
Emergency of International Concern (PHEIC)
We present here analytical studies and a method comparison
with clinical samples of a qualitative diagnostic real-time PCR assay,
the VERSANT® Zika RNA 1.0 Assay (kPCR)
Method: The VERSANT Zika RNA 1.0 Assay (kPCR) qualitatively
detects ZIKV RNA. ZIKV RNA from plasma or serum is extracted
using theVERSANTMolecular Prep SPwithVERSANT Sample Prepa-
ration 1.0 Reagents Kit and then amplified on the Thermo Fisher
QuantStudio 5 thermal cycler, Bio-Rad CFX96 Real-Time PCRDetec-
tion System, or the Applied Biosystems 7500 Real-time PCR System.
Two amplification reactions, targeted to portions of the NS2 and
NS5 regions of ZIKV, comprise the assay.
Inclusivity of the assay was evaluated in silico comparing
primer/probe sequences against 35 unique Zika sequences from
14 countries. Assay specificity was tested with 50 individual ZIKV-
negative clinical plasma specimens. Cross-reactivity of the assay
was evaluated with high titer inactivated pathogens: Dengue
(strains 1–4), Yellow Fever 17D, Chikungunya, West Nile, Human
Parvovirus B19, and Mayaro viruses, as well as protozoan Plas-
modium falciparium . An additional 51 organisms were evaluated
in silico. Analytical sensitivity of the assay was evaluated using
a dilution series of ZIKV (Zeptometrix, strain PRVABC59) with
concentration determined by a TCID50 endpoint dilution assay. A
method comparison with a CDC assay using primers/probe from
Lanciotti et al (2007) was conducted on 90 clinical plasma or serum
specimens suspected by a physician or confirmed by home brew
assay to contain ZIKV.
Results: All 35 of the published ZIKV strain sequences showed
100% homologywith at least one of the two amplification reactions.
No amplificationwas observed in ZIKV-negative clinical specimens.
No cross-reactivity was observed with any of the tested pathogens
tested and no significant sequence homology was found for any of
the 51 organisms evaluated in silico. An assay limit of detection of at
least 0.05U/mL (TCID50) was established in both plasma and serum
on each of the three thermal cyclers. In the method comparison,
both assays detected ZIKV in each of 34 specimens and did not
detect ZIKV in each of 44 specimens. The VERSANT Zika RNA 1.0
Assay (kPCR) detected ZIKV in an additional 10 specimens that the
comparator CDC assay did not, while the comparator CDC assay
detected ZIKV in 2 specimens that VERSANT Zika RNA 1.0 Assay
(kPCR) did not.
Conclusion: The VERSANT Zika RNA 1.0 Assay (kPCR) qualita-
tively detects ZIKV RNA. This assay recognizes a broad spectrum
of published ZIKV RNAs in silico, has high analytical sensitivity,
is specific to ZIKV among the family Flaviviridae viruses, and has
excellent performance with clinical specimens.