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

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Abstracts / Journal of Clinical Virology 82S (2016) S1–S142 S39
Abstract no: 267
Presentation at ESCV 2016: Poster 36
Comparison of respiratory and
Meningitis/Encephalitis viruses detected by
FilmArray® multiplex PCR versus real-time PCR
Roger Koller ∗, M.T. Barbani, A.U. Lüthi, S. Zürcher,
J.F. Steinlin-Schopfer, S.L. Leib,
M. Gorgievski-Hrisoho
Institute for Infectious Diseases, University of Bern,
Switzerland
Introduction: Fast and reliable pathogen detection is important
for adequate management of infections. Although real-time PCR
(rtPCR) is usually the most sensitive method for direct pathogen
detection, it requires experienced technicians, includes several
working steps and has a turnaround time of multiple hours. There-
fore this method is not ideal for emergency diagnostics. The FDA
cleared, fully automated sample to answer, FilmArray® (FA) mul-
tiplex PCR system (BioFire/bioMérieux) detects a broad spectrum
of pathogens in ∼70min. To optimize our diagnostic services
during weekends and off-peak times, we compared the FA Respira-
tory Panel (RP) and FA Meningitis/Encephalitis (ME) Panel to our
routinely used rtPCR assay. The FA panels detect 20 respiratory
pathogens (17 viruses, 3 bacteria) in nasopharyngeal swabs (NPS)
and 14M/E pathogens (7 viruses, 6 bacteria, Cryptococcus neofor-
mans/gattii) in cerebrospinal fluids (CSF).
Materials and methods: With FA we tested 84 retrospective
samples (23 NPS, 29 broncheoalveolar lavages [BALs], 32 CSF) and
60 prospectively collected NPS that required urgent testing during
the 2015/2016 flu season by FA and rtPCR. FA sample input volume
was 300ml for RP and 200ml forME. Commercial RP andME quality
control panels (MMQC Inc., Scarborough, USA), containing samples
positive and negative for each analyte detected by the FA panels,
were tested multiple times. For rtPCR, nucleic acids were extracted
from 220ml of sample and eluted in 55ml using NucliSENS easy-
MAG (bioMérieux). Respiratory viruses were analyzed by real-time
PCR using a combination of 7 duplex Respiratory Multi Well Sys-
tem r-geneTM (RG) assays (influenza A/B, RSV/hMPV, HRV&EV/cell
control, ADV/HBoV, HCoV/HPIV1-4) (Argene/bioMerieux), accord-
ing to manufacturer’s instructions. Additionally, we expanded FA
RP testing to include (BALs), by implementing one additional sam-
ple preparation step. CSF was analyzed for virus using laboratory
developed tests (LDTs) certified by the Swiss authorities.
Results: RP andME quality control panel results were 100% con-
cordant with expected results. For all NPS, both tests, FA RP and
RG, identified one or more viruses in 45/83 (54.2%) samples. FA
RP and RG results correlated for 42/48 viruses detected (87.5%).
FA RP detected an additional 3 HRV/EV and RG detected addition-
ally 1 FluA, 1 ADV and 1 HRV/EV. Positive percent agreement (PPA)
between RG (laboratory standard) and FA RP for NPS was 93.3%
and negative percent agreement (NPA) was 92.7%. Overall correla-
tion was 93.2%. Results from BALs yielded 92% PPA, 93.1% NPA and
overall correlation of 92.4%. For FA ME testing, 31/33 CSF samples
had identical FA ME and LDT results with an overall correlation of
94.4%. FA ME did not detect 2 parechovirus low level LDT positive
samples (Ct 36.3 and 37.0). Using LDTs as the laboratory standard,
FA ME PPA and NPA were 93.9% and 100%, respectively.
Conclusion: Results obtained with the FilmArray® RP and ME
panels were highly concordant with our currently used diagnos-
tic methods, demonstrating excellent performance. The simplicity
of the FilmArray® system, requiring less than 5min of hands-on
time, easy to read reports, and lowsample volume allows for testing
during off shifts and when urgent results are required. The compre-
hensiveness of the FilmArray® panels is ideal for diagnosing clinical
syndromes where there are many potential causes.
Abstract no: 273
Presentation at ESCV 2016: Poster 37
Cross-contamination and carry-over study
results obtained with ELITe InGenius, a new
sample-to-result solution for in vitro
diagnostics
C. Bittoto∗, S. Costa, M. Enrietto, S. Patanè,
F. Gorreta, A. Estampes, C. Olivo, G. Stefanuto,
N. Scarr, W. Mahoney
ELITechGroup Molecular Diagnostics, United
Kingdom
Background: ELITe InGeniusTM (ELITechGroup Molecular Diag-
nostics) is a fully automated sample-to-result solution, designed
for the in vitro molecular diagnostics and monitoring of infec-
tious diseases. The system combines on a single platform: sample
processing (extraction and purification), PCR set-up, real-time
amplification and detection of multiple parameters for qualita-
tive and quantitative analysis, and result interpretation. Absence
of cross-contamination and cross-over was evaluated within
robustness study.
Material/methods: ELITe InGenius features a universal extrac-
tion in a unitary cassette based format (ELITe InGenius SP200) and
multiple and independent Real-Time PCR with mixed parameters
including CE-IVD Real-Time PCR assays (ELITe MGB line) and Lab-
oratory Developed Tests. One to twelve patient samples can be
processed in 12 parallel trackswithin one run. Cross-contamination
and cross-over study protocols were designed in accordance with
FDA Draft Guidance for Industry and Food and Drug Administra-
tion Staff Establishing the Performance Characteristics of Nucleic
Acid-Based In vitro Diagnostic Devices. The tests to evaluate the
rate of false positive included: (1) 30 high positive MRSA sam-
ples (107 CFU/ml, prepared by dilution of MRSA BAA-1720 strain)
tested alternating to 30 negative samples within five 12-sample
runs with MRSA/SA ELITe MGB kit; (2) negative MRSA samples
(n = 50) tested within five runs along with 1 positive and 1 negative
MRSA/SA controls per run with MRSA/SA ELITe MGB kit, and (3) 30
high CMV positive samples (104 IU/ml, prepared by dilution of the
“1st WHO International Standard for Human Cytomegalovirus for
Nucleic Acid Amplification Techniques”) tested alternating to 30
negative samples within five 12-sample runs with CMV ELITe MGB
kit.
All samples were tested carrying out the whole analysis
procedure: extraction, amplification, detection and result interpre-
tation with ELITe InGeniusTM in combination with ELITechGroup
reagents.
Results: All negative and all positive MRSA and CMV samples
were correctly identified by the system. 100% of concordance with
the expected results was obtained for all the samples tested.
Conclusions: The results obtained demonstrated the total
absence of carry-over and cross-contamination of the system
even when high positive samples were tested. They confirm the
robustness and the reliability of ELITe InGenius for in vitro Molec-
ular Diagnostics testing.