Next Generation Panel Oriented Diagnostics The FilmArray Pathogen Identification System

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Next Generation Panel Oriented Diagnostics The
FilmArray Pathogen Identification System

• Idaho Technology

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The FilmArray
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The FilmArray is
User-Friendly Multiplex PCR
Automated Protocol
Integrated Sample Prep
Automated Results Analysis
Multiplex PCR
Speed
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Run Set-up
Load Pouch
Inject pouch Hydration Solution
Add sample to Sample Buffer
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Run Set-up
Inject sample
Load pouch in FilmArray
Enter pouch and sample ID, user info, then press
go!
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Automated Protocol
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Automated Protocol
5 min of hands on time
Start the run walk away
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Integrated Sample Prep
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Multiplex PCR
Simultaneous detection of dozens of targets
from one patient sample
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Automated Results Analysis
Positive or negative call for each target
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Automated Results Analysis
Eliminate Subjectivity
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Speed. Its really fast.
Sample prep
1 hour
RT-PCR
Multiplex PCR
Analysis
Results
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The FilmArray How does it work?
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The FilmArray Pouch
Reagent Storage
Chemical Circuit Board
Sample extraction purification
2nd stage PCR
1st stage multiplex PCR
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The FilmArray How Does it Work?
FilmArray Animation
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Automated Protocol

• Bladders inflate over blisters to push liquid

Air Channels

• Pistons close channels

Pneumatic Bladders
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The FilmArray Video analysis
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Integrated Sample Prep



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Nested MultiPlex PCR
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Automated Results Analysis
102 individual 2nd stage PCR wells Each well
contains one reaction Fluorescent data is
generated for each well
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Automated Results Analysis
All targets tested in triplicate
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Automated Results Analysis
For some organisms we target multiple genes
each tested in triplicate Software generates an
automated call for each organism
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The FilmArray Respiratory Panel
Bacterial Bordetella pertussis Chlamydophila
pneumoniae Mycoplasma pneumoniae

• Viral
• Adenovirus
• Bocavirus
• Coronavirus 229E
• Coronavirus HKU1
• Coronavirus OC43
• Coronavirus NL63
• Entero Virus
• Influenza A
• Influenza A H1
• Influenza A H1 2009
• Influenza A H3

Influenza B Metapneumovirus Parainfluenza 1
Parainfluenza 2 Parainfluenza 3 Parainfluenza
4 Respiratory Syncytial Virus Rhinovirus
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Nested Multiplex PCR
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Comparison of Multiplex PCR
Blue Graph - 5000 x LOD C. burnetti Red Graph -
0.1x LOD B. anthracis (100 ul of a 100 CFU/ml BA
sample)
LOD 1000 cfu/ml
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Robust Multiplex PCR
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Triple Infection
Bocavirus black PIV2 blue Human Rhinovirus -
green

• Bocavirus black
• PIV2 blue
• Human Rhinovirus - green

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Simultaneous Detection of 12 Organisms
Flu A H1 Flu A H3 Flu B RSV Para Flu 1 Para Flu 2
Para Flu 3 Adeno Human Meta Pneumo Rhino Boca
S. pneumoniae
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System Controls
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Internal pouch Controls

• Controls currently in the pouch
• RNA Process mRNA from freeze-dried S. pombe
• Pouch pass/fail is set on this control
• 1st Stage PCR synthetic oligos in PCR1
• 2nd Stage PCR synthetic oligos in PCR2
• Useful for understanding failure modes
• Human mRNA assay for human mRNA in sample
• We are collecting data

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Location of the controls
PCR2 control (spotted with PCR2 primers)
PCR1 control Freeze-dried with Primer Pellet
Yeast control Freeze dried into sample well
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Comparative Data
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The FilmArray Data
FilmArray vs DFA in 387 Pediatric NPAs
Pediatric NPA samples were tested by DFA (blue)
and by FilmArray (maroon).
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82 of 387 pediatric NPA samples tested positive
by FilmArray analysis
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32 tested positive for 2 or more pathogens by
FilmArray analysis
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Future Developments Product Development
(Beyond RP)

• Sepsis/Meningitis Panel
• Enteric Pathogens Panel
• STD Panel
• Biothreat Panel

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Questions