next generation panel oriented diagnostics the filmarray pathogen identification system idaho...

NEXT GENERATION PANEL ORIENTED DIAGNOSTICS THE FILMARRAY PATHOGEN IDENTIFICATION SYSTEM

Idaho Technology

The FilmArrayThe FilmArray

The FilmArray is…The FilmArray is…

User-Friendly Multiplex PCRUser-Friendly Multiplex PCR

Automated ProtocolAutomated Protocol

Integrated Sample PrepIntegrated Sample Prep

Multiplex PCRMultiplex PCR

Automated Results AnalysisAutomated Results Analysis

SpeedSpeed

Load Pouch

Run Set-upRun Set-up

Inject pouch Hydration Solution

Add sample to Sample

Buffer

Run Set-upRun Set-up

Inject sample

Enter pouch and sample ID, user info, then

press go!

Load pouch in FilmArray

Automated ProtocolAutomated Protocol

Automated ProtocolAutomated Protocol

5 min of hands on time

Start the run &Start the run &

walk awaywalk away

Integrated Sample PrepIntegrated Sample Prep

Multiplex PCR

Simultaneous Simultaneous detection of detection of dozensdozens of targets of targets from one from one patient samplepatient sample

Automated Results AnalysisAutomated Results Analysis

Positive or negative Positive or negative call for each targetcall for each target

Automated Results AnalysisAutomated Results Analysis

Eliminate SubjectivityEliminate Subjectivity

Speed…. It’s really fast.Speed…. It’s really fast.

Sample prep

Results

Multiplex PCR

Analysis

RT-PCR 1 hour1 hour

The FilmArray

How does it work?

The FilmArray PouchThe FilmArray Pouch

Reagent Reagent StorageStorage

Chemical Chemical Circuit Circuit BoardBoard

Sample Sample extraction & extraction & purificationpurification

11stst stage stage multiplex multiplex

PCRPCR

22ndnd stage stage PCRPCR

The FilmArray – The FilmArray – How Does it Work?How Does it Work?

FilmArray Animation

Automated Protocol

Pneumatic Bladders

Bladders inflate over blisters to push liquidAir Channels

Pistons close channels

The FilmArray: Video analysisThe FilmArray: Video analysis

Integrated Sample PrepIntegrated Sample Prep

+ + =

Nested MultiPlex PCRNested MultiPlex PCR

Automated Results AnalysisAutomated Results Analysis

102 individual 2nd stage PCR wells

Each well contains one reaction

Fluorescent data is generated for each well

Automated Results AnalysisAutomated Results Analysis

All targets tested in triplicate

Automated Results AnalysisAutomated Results Analysis

For some organisms we target multiple genes – each tested in triplicate

Software generates an automated call for each organism

23

The FilmArray Respiratory Panel

ViralAdenovirus Bocavirus Coronavirus 229E Coronavirus HKU1 Coronavirus OC43 Coronavirus NL63 Entero VirusInfluenza A Influenza A H1Influenza A H1 2009Influenza A H3

Bacterial Bordetella pertussis Chlamydophila pneumoniae Mycoplasma pneumoniae

Influenza B Metapneumovirus Parainfluenza 1 Parainfluenza 2 Parainfluenza 3 Parainfluenza 4 Respiratory Syncytial Virus Rhinovirus

Nested 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)

Comparison of Multiplex PCR

*LOD = 1000 cfu/ml

Robust Multiplex PCR

Triple Infection

Bocavirus – black

PIV2 – blue

Human Rhinovirus - green

Bocavirus – black

PIV2 – blue

Human Rhinovirus - green

Simultaneous Detection of 12 OrganismsSimultaneous Detection of 12 Organisms

Flu A H1Flu A H3Flu BRSVPara Flu 1Para Flu 2

Para Flu 3AdenoHuman Meta PneumoRhino Boca S. pneumoniae

System 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

Internal pouch Controls

Location of the controls

PCR2 control (spotted with PCR2 primers)

PCR1 controlFreeze-dried with Primer Pellet

Yeast controlFreeze dried into sample well

Comparative Data

33

The FilmArray – Data

FilmArray vs DFA in 387 Pediatric NPAs

Pediatric NPA samples were tested by DFA (blue) and by FilmArray (maroon).

82% of 387 pediatric NPA samples tested 82% of 387 pediatric NPA samples tested positive by FilmArray analysispositive by FilmArray analysis

32% tested positive for 2 or more 32% tested positive for 2 or more pathogens by FilmArray analysispathogens by FilmArray analysis

Future Developments

Product Development (Beyond RP)

Sepsis/Meningitis Panel

Enteric Pathogens Panel

STD Panel

Biothreat Panel

Questions