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

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S120 Abstracts / Journal of Clinical Virology 82S (2016) S1–S142
NAA was performed using the MUNANA substrate obtained
from both Biosynth AG and Sigma-Aldrich companies. The NIA was
conducted for both MUNANA brands to assess the susceptibilities
of all IV to oseltamivir and zanamivir.
Additionally, NIA was performed for each IV strain with alter-
native sources of NAIs: oseltamivir – F. Hoffmann-La Roche Ltd vs.
oseltamivir – Sequoia Research Products Ltd (SRP); zanamivir –
GlaxoSmithKline vs. zanamivir – SRP; laninamivir – Daiichi Sankyo
vs. laninamivir – SRP and peramivir – BioCryst Pharmaceuticals.
Statistical analyses were performed by one-way analysis of vari-
ance (ANOVA) followed by Tukey’s Honestly Significant Difference
post-hoc test ( ˛ = 0.05) using R.
Results: No statistically significant difference was established
for the IC50 means of each pair of commercially available reagents.
MUNANA from both companies performed similarly in deter-
mining NA activity of the selected IV and exhibited the same profile
in determining the IC50mean values for oseltamivir and zanamivir.
IC50 values of the selected IV determined for oseltamivir,
zanamivir and laninamivir exhibited the same potency for the dif-
ferent sources of NAIs.
Conclusions: MUNANA substrate compound and NAIs previ-
ously available from a single supplier can now be purchased from
other chemical companies and at a significantly low price. Given
the limited resources of research and public health funding, prefer-
ence for alternative suppliers might be translated into cost savings
or low bureaucratic nuisance. This strategy may maximize funding
resources and allow researchers to divert more funds to targeted
research goals. Laboratories are encouraged to consider these cost-
efficient alternative suppliers as a reliable solution.
Abstract no: 278
Presentation at ESCV 2016: Poster 201
Evaluation of point of care testing platform
(ePLEX) for respiratory viral diagnosis
Daniel Guerendiain ∗, Laura MacKenzie,
K.E. Templeton
NHS Lothian, Royal Infirmary of Edinburgh, UK
Background: There is an increasing demand on laboratories to
deliver respiratory viral diagnosis by molecular methods. Different
strategies are explored which include – point of care testing which
is simple, requires minimal hands on time, is fast and can be done
in ward areas and not in centralised laboratories. Tests considered
need to be shown to have good performance.
GenMark Diagnostics Inc. (Carlsbad, USA) has developed a respi-
ratory panel assay (RP) for the ePlex system detecting 26 microbes,
including 22 virus and 4 (atypical) bacteria in 90min.
The RP cartridge contains all reagents required to run the RP
Panel assay. Lysis and nucleic acid extraction, PCR amplification
andhybridization-based electrochemical detection occur inside the
cartridge, reducing the hands-on-time to less than 1min per sam-
ple.
The objective of this study was to compare and study the per-
formance of the new GenMark ePlex assay against the in house
real-time PCR, a lab developed test (LDT).
Material and methods: 81 nasopharyngeal swabs samples
(NPS) in UTMwere previously tested by an in-house Real Time PCR.
Samples selected contained the following respiratory pathogens:
respiratory syncytial virus, influenza A, influenza B, rhinovirus,
enterovirus, bocavirus, coronaviruses, metapneumovirus, parain-
fluenza viruses, Bordetella pertussis and Mycoplasma pneumoniae
32.1% samples were co-infected, even with 4 different organisms.
Samples selected were less than 4 months old with only one
freeze/thaw cycle. Ct values ranging from 17.11 to 40.39 mean
24.74.
200 l of each NPS sample was added to the ePlex Sample Buffer
device, transferred to the RP cartridge and then inserted on the
ePlex device.
Agreement between the original LDT results and the results
obtained with the ePlex assay was assessed as detected or not
detected.
Additionally 5 successive 1:10 dilutions were performed for 7
different specimens: RSV, influenzaAH1N1, influenza B, rhinovirus,
bocavirus, Mycoplasma pneumoniae and Bordetella pertussis. Dilu-
tions were tested on both assays to identify and compare the lower
limit of detection to the LDT.
Results: Total concordance was observed in 91.73% of cases.
Only 10 discrepancies were identified. 7 organisms were detected
by the ePlex assay andmissed by the LDT and 3 organismswere pos-
itive detected by the LDT and negative by the ePlex. Discrepancies
were repeated in both assays showing same results.
Total concordance was observed in 80% of dilutions. 1 dilution
10−4 RSV sample was detected by the ePlex and resulted negative
for the LDT. As well 6 samples (10− 3–10 −5 dilutions) were positive
for the LDT and negative for the ePlex assay. B. pertussis did not
detect the 2 lower dilutions as a different target gene was in use in
ePlex assay.
Conclusion: The preliminary evaluation on a small sample set
show a very good agreement across a range of pathogens with the
GenMark compares in house real-time PCR. The assay was also
found to be very simple and easy to perform and would be suitable
for a hospital ward or outpatient environment.
Abstract no: 286
Presentation at ESCV 2016: Poster 202
Single genetic clades of EV-D68 strains in 2010,
2013, and 2015 in Osaka City, Japan
A. Kaida ∗ . Iritani . Yamamoto
D. Kanbayashi , Y. Hirai , U. Kohdera ,
M. Togawa Amo Shiomi ishigaki
T. Kageyama , H. Kubo
1 Osaka City Institute of Public Health and
Environmental Sciences, Japan
2 Nakano Children’s Hospital, Japan
3 Osaka City General Hospital, Japan
4 Aizenbashi Hospital, Japan
5 Osaka Police Hospital, Japan
6 Influenza Virus Research Center, National Institute
of Infectious Diseases, Japan
Background: Detection of Enterovirus D68 (EV-D68), a cause
of acute respiratory tract infection (ARTI), was rarely reported
before the early 2000s. Molecular analyses have demonstrated that
recently detected EV-D68 strains are of three major genetic clades.
We previously reported the emergence of EV-D68 in children with
ARTI in Osaka City, Japan in 2010.
Objectives: This study surveyed EV-D68 among children with
ARTI since its first endemic period in 2010 and conductedmolecular
analyses of the detected viral genome sequences.
Methods: During November 2010–December 2015, 2215 respi-
ratory clinical specimens were obtained from children (<10 years
old) with ARTI. Specimens from patients diagnosed with influenza
were excluded. Real-time RT-PCRwas used to detect enteroviruses.
Viral protein 4 (VP4) or VP1 genes were sequenced to identify EV-