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

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S50 Abstracts / Journal of Clinical Virology 82S (2016) S1–S142
Results: The data from2015was anonymised, collated, and out-
liers removed. The data were split into categories based on the
assay manufacturer who provided the amplification reagents. Out-
liers were assessed through the application of Grubbs’ analysis to
each assaymanufacturing group. Datasetswere considered suitable
for inclusion if more than one laboratory reported data using the
same assay manufacturer after removal of non-compliant data and
outliers. For CMV, 120 quantitative datasets across 5 assays were
returned, 106 for EBV, 132 for JCV and 144 datasets for BKV. Assay
manufacturing group datasets were compared against the group
consensus and the dPCR assigned values.
Conclusions: The comparison of inter-laboratory data for viral
load determination is limited in the absence of certified control
materials or international standards due to potential assay vari-
ation. The aim of this study was to compare the quantitative
performance of dPCR against the key commercial assays available
for the four viral targets. The results showed that the dPCR results
aligned closelywith the quantitative values determined using com-
mercial assays and the in-house qPCR assay independent of an
International Standard being available. Historical assay data (not
shown) support this data and show the close alignment between
qPCR and dPCR assays. Digital PCR allows quantitation without
the need for a calibrated standard and from the preliminary data
presented here indicates a method by which to calibrate controls
allowing comparison of results between laboratories.
Abstract no: 71
Presentation at ESCV 2016: Poster58
Comparison of two multiplexed PCR assays for
respiratory virus detection in ICU patients:
FilmArray® respiratory panel and AllplexTM
respiratory full panel
M. Gozalo-Margüello ∗, I. Angulo-López ,
A. Aguirre-Qui˜nonero , C. Ruiz de Alegría ,
J. Agüero-Balbín , L. Martínez-Martínez
1 University Hospital Marqués de Valdecilla-IDIVAL,
Spain
2 University Hospital Marqués de Valdecilla-IDIVAL.
Department of Molecular Biology. University of
Cantabria, Spain
Background and objectives: The FilmArray® Respiratory Panel
(BioFire Diagnostics, LLC, Salt Lake City, UT, USA, a bioMérieux
Company) is a multiplexed automated PCR assay that integrates
specimen processing, nucleic acid amplification, and detection into
a pouch, and detects 17 respiratory viruses plus three bacteria. We
compared it to another multiplexed PCR assay, AllplexTM Respira-
tory Full Panel (Seegene, Inc., Seoul, Korea) composed of 4 different
panels in a multiplex One-step Real-time RT-PCR assay to detect
and identify 16 respiratory viruses and 7 bacteria in patient’s spec-
imens. We conducted a study to evaluate the performance of the
FilmArray® compared to the AllplexTM for the detection of respira-
tory viruses from different respiratory specimens in ICU patients.
Bacteria were not taken into account in this study.
Methods: A prospective comparative study was carried out in
50 respiratory specimens (nasal swabs, nasopharyngeal lavage,
bronchoalveolar lavage, bronchoaspirate and sputum) collected
from ICU patients between March and May 2016. One aliquot
was processed with the Allplex TM and a second aliquot was
tested by the FilmArray® assay. For the AllplexTM assay, viral
nucleic acid was extracted using the BioRobot EZ1® (Qiagen).
Both assays detect influenza A (Flu A; seasonal H1, subtype 2009
Table 1
Discrepant results between FilmArray ® Respiratory Panel and AllplexTM Respiratory
Full Panel, and virus-specific RT-PCR results.
Sample FilmArray®
respiratory panel
AllplexTM respiratory
full panel
Specific RT-PCR
Nasal swab AdV Negative Negative
Nasal swab Flu A 2009 Negative Flu A 2009
Nasal swab AdV Negative Negative
Nasal swab Flu A 2009 Negative Flu A 2009
Sputum Negative Flu B Negative
Nasal swab Negative Flu B Negative
Nasal swab Negative AdV AdV
BAL Negative AdV AdV
H1 and H3) and influenza B (Flu B), respiratory syncytial virus
(RSV), adenovirus (AdV), human rhinovirus/enterovirus (HRV/E),
parainfluenza 1-4 (PIV 1-4), humanmetapneumovirus (HMPV) and
coronaviruses (CoV) NL63/229E/OC43. Additionally, FilmArray®
detects CoV HKU1 and AllplexTM, bocavirus (BoV). For discrepant
results, virus-specific RT-PCR was performed (RealStar® RT-PCR
Kits, Altona Diagnostics, Hamburg, Germany).
Results: Of the 50 specimens tested, both assays agreed on 21
negative and 21 positive respiratory specimens. Discrepant results
(8) agreed with specific PCR in 4 for FilmArray® and in 4 for
AllplexTM The FilmArray® showed a 92.6% sensitivity and
91.3% specificity compared to 92.3% sensitivity and 88.5% specificity
for AllplexTM.
Conclusions: The FilmArray® is a useful, easy-to-perform assay
for detecting respiratory viruses in ICU patients. The AllplexTM
requires additional RNA extraction time and more hands-on time,
making it longer to perform. However, as the FilmArray® processes
one single sample at a time, this increases the time of successive
results when several samples arrive to the laboratory with a short
difference in arrival time, but this point is solved by new platforms
available.
Abstract no: 76
Presentation at ESCV 2016: Poster 59
Evaluation of three different sample
populations on a new multiplex BioPlex®2200
assay for the detection of measles, Mumps, and
Varicella-Zoster virus IgM antibodies
D. King ∗, R. Del Rosario, C. Tatyosian, C. Todd,
J. Flanagan, J. Vogel
Bio-Rad Laboratories, Hercules, CA, United States
Background: Measles, mumps, and varicella are three of the
most highly infectious diseases. Worldwide outbreaks continue in
many countries despite aggressive vaccination campaigns. Early
diagnosis of these diseases improves patient management and
helps prevent outbreaks from spreading. To assist clinicians in
making quick and accurate diagnoses, Bio-Rad Laboratories is
developing a new assay used for the identification of IgM class
antibodies to measles, mumps, and varicella-zoster virus (VZV) in
human serum or plasma. The new BioPlex 2200 MMV IgM assay
produces three discrete results from a single multiplexed test reac-
tion and is being developed to accommodate a wide variety of
sample types to facilitate diverse testing situations.
Methods: Retrospective samples positive by Diasorin Liaison
assays (measles n = 104, mumps n = 183, and VZV n = 64), a test
ordered sample population comprised of samples from a Euro-
pean reference laboratory (measles n = 300 mumps n = 300, and
VZV = 300), and samples froma healthy populationmade up of sam-