FilmArray(Biofire)Syndromic Panel

The fastest way to better results

Dr.Kavitha.M.P.
HOD & Consultant
Microbiologist,
Anand Diagnostic Laboratory,
Bangalore

1
 Order of Presentation
• Introduction of FilmArray.
• What are the panels available?
• Its clinical impact.
• How does FilmArray work?
• What are panels done at Anand Diagnostic Laboratory ?
• Conclusion

2
 Introduction
• The FilmArray - multiplex PCR system that integrates sample preparation,
amplification, detection and analysis.

• It is designed to be used with comprehensive panels that each offer testing for
sets of pathogens associated with some of today's MOST pressing healthcare
challenges

3
 One system. Many applications.
Comprehensive – Panels cover a wide range of targets involved in causing
respiratory, bloodstream, gastrointestinal and central nervous infections

Blood Culture Meningitis

Respiratory Gastrointestinal
Identification Encephalitis
Panel Panel
Panel Panel

20 27 22 15
targets targets targets targets
• 19 bacteria
• 13 bacteria • 6 bacteria
• 3 bacteria • 5 yeast
• 5 viruses • 8 viruses
• 17 viruses • 3 antibiotic
• 4 parasites • 1 fungi
resistance genes
 One system. Fully integrated

27/01/2022
5
 THE PANELS

FilmArray
 Respiratory Panel
20 pathogens

Viral
Parainfluenza 1
Adenovirus
Parainfluenza 2
Coronavirus 229E Parainfluenza 3
Coronavirus HKU1 Parainfluenza 4
Coronavirus OC43 RSV
Coronavirus NL63
Human Metapneumovirus
Human Rhinovirus/ Enterovirus Bacterial
Influenza A Bordetella pertussis
Influenza A/H1 Chlamydophila pneumoniae
Influenza A/H1-2009 Mycoplasma pneumoniae
Influenza A/H3
Influenza B

Sample : Nasopharyngeal Swabs

7
 Specimen Collection and Preparation

Nasopharyngeal swab (NPS) in Viral Transport Medium (VTM)

• 300µL required for testing

Specimen transport and storage

• Room temperature (18-30ºC) for up to 4 hours
• Refrigerated (2-8ºC) for up to 3 days
• Frozen (≤-15ºC) for up to 30 days
 Blood Culture ID Panel
27 pathogens

Gram + Bacteria: Gram - Bacteria : Fungi:

Enterococcus spp. A. baumannii C. albicans
L. monocytogenes Enterobacteriaceae C. glabrata
Staphylococcus Enterobacter cloacae C. krusei
S. aureus Complex C. parapsiolosis
Streptococcus spp. E. coli C. tropicalis
S. agalactiae (Group B)H. influenzae
S. pyogenes (Group A) K. oxytoca
S. pneumoniae K. pneumoniae
N. meningitidis
Antibiotic Resistance: P. aeruginosa
mecA Proteus
Van A/B S. marcescens
KPC
Sample : Positive Blood culture
 Lab workflow

Used on blood culture bottles

– Positively labelled by incubation systems
 GI Panel
22 pathogens
Bacteria: Protozoa:
Aeromonas Cryptosporidium
Campylobacter Cyclospora cayetanensis
Clostridium difficile (Toxin A/B) Entamoeba histolytica
Plesiomonas shigelloides Giardia lamblia
Salmonella
Vibrio Viruses:
Vibrio cholerae Adenovirus F 40/41
Yersinia enterocolitica Astrovirus
Norovirus GI/GII
Diarrheagenic E. coli / Shigella Rotavirus A
E. coli O157 Sapovirus
Enteroaggregative E. coli (EAEC)
Enteropathogenic E. coli (EPEC)
Enterotoxigenic E. coli (ETEC)
Shiga-like toxin-producing E. coli (STEC)
Shigella/Enteroinvasive E. coli (EIEC)
Sample : Stool resuspended in Cary Blair
 Specimen Collection and Preparation

• Stool in Cary Blair Transport Media

• Provides a liquid sample

• Add stool and mix

• Do not overfill
• Sample Volume – 200 µL

• Sample Storage
• Up to 4 days

• Ambient or refrigerator temperature

 Meningitis / Encephalitis
15 pathogens
Bacteria: Viruses:
E. coli Cytomegalovirus (CMV)
H. influenzae Enterovirus
L. monocytogenes Epstein-Barr virus (EBV)
N. meningitidis Herpes simplex type 1 (HSV-1)
S. agalactiae Herpes simplex type 2 (HSV-2)
S. pneumoniae Human herpesvirus 6 (HHV-6)
Parechovirus
Fungi: Varicella zoster virus (VZV)
Cryptococcus
neoformans / gattii

Sample : Cerebral Spinal Fluid

 Specimen Collection

Sample CSF

Volume 200 µL

Transport Ambient temperature

 Clinical Impact

 Targeted therapy with

adequate
antiviral/antifungal  Fast and targeted patient
Economic impact
and/or antibiotics isolation
 Less secondary testing
 Faster therapeutic  Preventing spread of
pathogens  Less secondary
adjustment interventions
 Improved patient  Reduction in length of stay
 Length of stay reduction
management  Rapid outbreak identification
 Appropriate antibiotic tool
use in critical care
 HOW DOES FILMARRAY WORK

FilmArray
Integrated Sample Preparation, Amplification, and Detection
 Setting Up the FilmArray Is Easy

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2 minutes of
hands-on time

6.
5.4. 2.
The
The
The
Hydration
sample/buffer
lid
sample
of theissolution
sample
added
mixture
injection
to
is injected
theis sample
injected
vial
intois injection
closed
the
intopouch
the
and
vial
pouch
the
through
using
vial
through
the
isthe
inverted
transfer
blue
the red
3
3. The
1.
Sample
The
FilmArray
pouch
bufferinstrument
is inserted
added tois into
the
now the
sample
ready
loading
injection
to set-up
blockvial
times toinlet
inlet
pipette
mixport
port
the sample
Sample Extraction, Amplification, and Detection: It’s All in the Pouch

The FilmArray pouch is loaded into the FilmArray instrument

 Sample extraction, amplification, and
detection: It’s all in the pouch

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65 minutes
run-time

6.4.2.
5. 1. Nucleic
Nucleic
The acids
8.3.First-stage
Products
Sample acids
An FilmArray
elution
from
moves move
bound
PCRbuffer
the
products
into to
performs the
by
lysis afirst-stage
magnetic
first-stage
removes
are
chamber.
added
PCR
purified
melt are
to to PCR
beads
Cells chamber.
move
diluted
nucleic
fresh
confirmand from
master
to Reverse
pathogens
acids
the removetheand
from
mix
presence lysis
any
are
the transcriptase
or arechamber
remaining
lysed
magnetic to
aliquoted
absence by bead
ofPCR
converts
the
primers
into
beating,
beads
each target
purification
well RNA
releasing
assay-specific to DNA,
chamber.
oftemperature
the nucleic
array A followed
wash
acids
signatures by
buffer athe
high-order
ofremoves
second multiplex
cellular
stageand PCR
PCRpathogen
product
7. Each well is pre-spotted with a single pair of second-stage PCR primers,
debris
for each well in the array
resulting in specific amplification of target DNA only. A fluorescent double-
stranded DNA binding dye monitors each reaction
 2 internal controls

• RNA Process Control

Freeze-dried Schizosaccharomyces pombe organism
Re-suspended with specimen
• PCR II Control
DNA template spotted on the array

PCR II
RNA Control
Process
Control
 Automated Results Analysis
• All targets tested in triplicate
• Two out of three wells must be positive
• Melting peaks must fall in their specific range
• Melting peaks must be significantly similar to each other
• Both internal controls must pass (RNA process and PCR 2)

Bordetella
pertussis
 PANELS AVAILABLE AT ANAND DIAGNOSTIC LABORATORY

FilmArray
 Panels at Anand Diagnostic Laboratory

1.Blood panel
2.Respiratory panel
3.Gastrointestinal panel
4.Meningitis panel
 Identification
PANEL Detected

BCID 1.Staphylococcus aureus Mec A gene

Respiratory 1.Human
Rhinovirus/Enterovirus
2.Influenza A H3
3.Adeno virus
4.RSV
GI 1.Vibrio cholerae
2.ETEC lt/st
3.Salmonella/EAEC
4.Rota virus
5.C.difficile/EPEC/ETEC
6.Shigella/EIEC/
Cryptosporidium/Norvovirus
7.Giardia
ME 1.Cryptococcus
neoformans/gattii
2.Streptococcus pneumoniae
3.HSV 1
4.CMV
 Case Presentation-GI panel

A boy aged 11 years,diagnosed AML, Post bone marrow transplant was

admitted with history of 4 episodes of loose stools. To the working diagnosis of
infective gastro enteritis versus gut GVHD, he was started on IV fluids, antibiotics
(Metronidazole) after sending blood, urine and stool cultures.
Biofire diarrhea panel was sent and it showed Shigella /E.coli EIEC,
Cryptosporidium and Norvovirus .

The diarrhea was not suggestive of GVH gut with respect to  volume ( <500ml/
no colic/ blood) and consistency. In view of positive infective etiology accounting
for large bowel type of diarrhea, a flexible sigmoidoscopy and biopsy  to exclude
GVH gut was deferred.

He was started on Ceftriaxone, Metronidazole and Nitazoxamide in consultation 

with ID specialist. All cultures were sterile.  He remained afebrile during his
hospital stay. He responded well to  anti-infective therapy. Stools are currently
formed and there were no abdominal signs at discharge.
 Case Presentation ME panel
44 days old, term baby with neonatal seizures was referred to higher center.

MRI report showing subacute infarction in right frontal lobe with focal IVH,B/L
ventricles.
O/E:
No jaundice , Enlarged spleen and Liver
CSF-Grams stain and Culture-Negative
Blood culture-Negative
HSV 1 & 2 PCR-Negative
CRP-Negative

CSF sent for FilmArray : CMV detected

Started on Gancyclovir , baby responded well, shifted to ward and discharged by
end of 1 week.
 Case presentation Respiratory Panel
New born , Preterm/LBW with Pneumonia was admitted with C/O Cough,
Cold and Fever since 5 days.
Respiratory distress, refusal of feeds
O/E:
B/L crepts

CRP-Negative
TC-8,850
Blood culture-Negative
H1N1-Negative

Nasopharyngeal swab sent for FilmArray : RSV detected

 Advantages and Disadvantages

Advantages
1.Easy and minimal sample
volume required
2.Reduction in TAT
3.No cross contamination
4.Detects multiple targets
5.The test run in triplicates
Disadvantages
1.Can run only one sample at a
time
2.Direct whole blood cannot
be used for BCID panel
 Conclusion

Film Array performs the extraction, amplification and detection in a

closed system minimizing contamination.
It is a syndromic testing system for a variety of pathogens associated
with key healthcare concerns.
The rapid results help physicians attain actionable information
sooner so appropriate decisions can be made.
The system is particularly relevant in complex diagnostic situations
that require answers in a critically short period of time.
 Conclusion

The FilmArray decreases the duration of antibiotic use, the length of

inpatient stay, and the time in isolation.
Impacted viral positive patients allowing earlier discontinuation of
antibiotics and reducing nosocomial infections.
The ease of use of the assay will likely help laboratories to adopt and
offer it 24/7,thus improving patient care.
The GI panel has the potential to more quickly identify and further
reduce transmission in an outbreak of enteric pathogens
THANK YOU