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

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S40 Abstracts / Journal of Clinical Virology 82S (2016) S1–S142
Abstract no: 279
Presentation at ESCV 2016: Poster 38
Validation of ELITe InGeniusTM, a flexible
sample-to-result solution, for viral meningitis
and encephalitis testing
C. Bittoto ∗, S. Costa, M. Enrietto, S. Patanè,
F. Gorreta, A. Estampes, C. Olivo, G. Stefanuto,
W. Mahoney
ELITechGroup Molecular Diagnostics, United
Kingdom
Background: ELITe InGeniusTM (ELITechGroup Molecular Diag-
nostics) is a new and fully automated cassette based sample-to-
result solution combining a universal extraction and independently
controlled real-time PCR thermal cyclers. The system was CE-
IVD validated for the rapid diagnostic and the monitoring of viral
meningitis based on the detection of Enterovirus, HSV1, HSV2 and
VZV with ELITe MGB assays in cerebrospinal fluid (CSF) patient
samples. This study describes the performance verification and
validation of these 4 assays on CSF samples and its advantages in
terms of workflow.
Material/methods: ENTEROVIRUS ELITe MGB® Kit, HSV1 ELITe
MGB® Kit, HSV2 ELITe MGB® Kit and VZV ELITe MGB® Kit (ELITech-
Group Molecular Diagnostics) are real-time PCR assays based on
MGB technology. ENTEROVIRUS ELITeMGB® Kit is a one-step assay:
reverse transcription of the viral RNA and Real-Time amplification
are performed in the same reaction. The four assays were validated
with ELITe InGenius TM using a universal extraction process in uni-
tary cassette based format (ELITe InGeniusTM SP 200) validated for
both viral RNA and DNA extraction. The CSF volume required was
200 L. The complete volume was processed by the system with
an internal control to check the integrity of the full process. The
extracted nucleic acids were eluted in 100 L in a specific collec-
tion tube. 20 L of extracted samples were then used for each DNA
based assays (HSV1, HSV2, VZV ELITe MGB assays) and 10 L for
the Enterovirus ELITe MGB assay. The validation study included:
(1) analytical studies to verify the PCR performances: efficiency,
linearity, accuracy, repeatability, reproducibility, sensitivity; and
(2) clinical study to assess the diagnostic sensitivity and speci-
ficity by testing with each assays: positive clinical and contrived
CSF samples (n = 20) and negative CSF samples (n = 22).
Results: The four real-time PCR assays passed the performance
acceptance criteria established for both analytical studies and
clinical studies on CSF samples. The global diagnostic sensitivity
obtained with the four ELITe MGB kits was 100% (80/80) and the
diagnostic specificity was 100% (88/88) on the CSF samples. In
terms of workflow, the storage of the extracted nucleic acids in
a dedicated tube and the independently controlled unitary thermal
cyclers enable the laboratory to perform multiple PCR in parallel
from one single CSF extracted sample, even with different thermal
profiles, or to perform the PCR reactions in separate runs with the
possibility to store the remaining eluate for retesting or archiving.
Conclusions: The results of the studies support the CE-IVD
marking of ELITe InGeniusTM in combination with Enterovirus,
HSV1, HSV2 and VZV ELITe MGB assays for viral meningitis and
encephalitis testing. The unique design of ELITe InGeniusTM enables
the laboratory to use single assay or to create custom panels com-
bining the four parameters according to their specific needs simply
on demand. The resulting flexibility associated to a short hands-
on time could contribute to improve the laboratory workflow for
viral meningitis testing. ELITe InGenius TM is the first sample-to-
result solutionwith a CE-IVDquantitativemenu for viral meningitis
testing.
Abstract no: 288
Presentation at ESCV 2016: Poster 39
Comparing performance of NxTAG RPP, xTAG
RVP Fast v2 and FilmArray RP for detecting
respiratory pathogens in nasopharyngeal
aspirates and swine/avian origin influenza A in
culture isolates
K.H. Chan ∗, P. Li, T.L. Wong, R. Chang, K.H. Chik
Department of Microbiology, The University of Hong
Kong, Hong Kong
Introduction: Rapid and accurate identification of the causative
pathogens of respiratory tract infections including influenza can
help guide treatment decisions with specific antiviral therapy,
implementation of infection control measure, possibly reducing the
length of hospital stay and associated healthcare costs.
Objective: This study is to compare the performance of NxTAG
RPP (CE-IVD) with xTAG RVP Fast v2 IVD and FilmArray for
detecting respiratory pathogens in nasopharyngeal aspirates, and
swine/avian origin influenza A in culture
Materials and methods: Seventy-one NPA samples collected
fromQueenMaryHospital patients suspectedwith respiratory viral
infections together with two swine and five avian origin influenza
A culture isolates: H1N1pdm09, H3N2 variant, H2N2, H5N1, H5N6,
H7N9 and H9N2 were recruited for this evaluation. Testing will be
performed according to the respective product inserts for NxTAG
RPP, xTAG RVP FASTv2 and FilmArray AP.
Results: The sensitivity, specificity, positive predictive value and
negative predictive value of NxTAG RPP, xTAG RVP and FilmArray
AP are 97.1%, 99.7%, 95.2% and 99.8%; 96.9%, 99.9%, 99.0% and 99.7%;
85.3%, 99.9%, 98.8% and 98.9% respectively. When NxTAG RPP was
compared with xTAG RVP and FilmArray RP, the concordance of
positive and negative results, Kappa is 0.95 (95% CI 0.91–0.98) and
0.92 (95% CI 0.87–0.98) respectively, while xTAG RVP compared
with FilmArray RP, Kappa is 0.90 (95% CI 0.85–0.95). All influenza
A subtype isolates were detected by matrix gene and the mean
of analytic sensitivity of NxTAG RPP, xTAG RVP and FilmArray RP
were 0.3, 12.3 and 0.5 of TCID50 respectively. The genotyping gene
remains negative except H1N1pdm2009 isolate. H3N2 variant was
subtyped as seasonal H3N2 by NxTAG RPP.
Conclusion: In this study, NxTAG RPP, xTAG RVP and FilmAr-
ray AP for detecting respiratory pathogens have high concordance
results between them. NxTAG RPPwas themost sensitive for detec-
tion of respiratory pathogens than NxTAG RVP and FilmArray RP.
Despite NxTAG RVP gives incorrect subtyping of H3N2 variant, it is
still the most sensitive assay for detection of avian or swine origin
Influenza A that threaten human life.