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NEXT GENERATION PANEL ORIENTED DIAGNOSTICS THE FILMARRAY PATHOGEN IDENTIFICATION SYSTEM Idaho Technology.

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Presentation on theme: "NEXT GENERATION PANEL ORIENTED DIAGNOSTICS THE FILMARRAY PATHOGEN IDENTIFICATION SYSTEM Idaho Technology."— Presentation transcript:

1 NEXT GENERATION PANEL ORIENTED DIAGNOSTICS THE FILMARRAY PATHOGEN IDENTIFICATION SYSTEM Idaho Technology

2 The FilmArray

3 The FilmArray is… User-Friendly Multiplex PCR Automated Protocol Integrated Sample Prep Multiplex PCR Automated Results Analysis Speed

4 Load Pouch Run Set-up Inject pouch Hydration Solution Add sample to Sample Buffer

5 Run Set-up Inject sample Enter pouch and sample ID, user info, then press go! Load pouch in FilmArray

6 Automated Protocol

7 5 min of hands on time Start the run & walk away

8 Integrated Sample Prep

9 Multiplex PCR Simultaneous detection of dozens of targets from one patient sample

10 Automated Results Analysis Positive or negative call for each target

11 Automated Results Analysis Eliminate Subjectivity

12 Speed…. Its really fast. Sample prep Results Multiplex PCR Analysis RT-PCR 1 hour 1 hour

13 The FilmArray How does it work?

14 The FilmArray Pouch Reagent Storage Chemical Circuit Board Sample extraction & purification 1 st stage multiplex PCR 2 nd stage PCR

15 The FilmArray – How Does it Work? FilmArray Animation

16 Automated Protocol Pneumatic Bladders Bladders inflate over blisters to push liquid Air Channels Pistons close channels

17 The FilmArray: Video analysis

18 Integrated Sample Prep ++ =

19 Nested MultiPlex PCR

20 Automated Results Analysis 102 individual 2 nd stage PCR wells Each well contains one reaction Fluorescent data is generated for each well

21 Automated Results Analysis All targets tested in triplicate

22 Automated Results Analysis For some organisms we target multiple genes – each tested in triplicate Software generates an automated call for each organism

23 23 The FilmArray Respiratory Panel Viral Adenovirus Bocavirus Coronavirus 229E Coronavirus HKU1 Coronavirus OC43 Coronavirus NL63 Entero Virus Influenza A Influenza A H1 Influenza A H Influenza A H3 Bacterial Bordetella pertussis Chlamydophila pneumoniae Mycoplasma pneumoniae Influenza B Metapneumovirus Parainfluenza 1 Parainfluenza 2 Parainfluenza 3 Parainfluenza 4 Respiratory Syncytial Virus Rhinovirus

24 Nested Multiplex PCR

25 Blue Graph 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

26 Robust Multiplex PCR

27 Triple Infection Bocavirus – black PIV2 – blue Human Rhinovirus - green Bocavirus – black PIV2 – blue Human Rhinovirus - green

28 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

29 System Controls

30 Controls currently in the pouch: RNA ProcessmRNA from freeze-dried S. pombe Pouch pass/fail is set on this control 1 st Stage PCRsynthetic oligos in PCR1 2 nd Stage PCRsynthetic oligos in PCR2 Useful for understanding failure modes Human mRNAassay for human mRNA in sample We are collecting data Internal pouch Controls

31 Location of the controls PCR2 control (spotted with PCR2 primers) PCR1 control Freeze-dried with Primer Pellet Yeast control Freeze dried into sample well

32 Comparative Data

33 33 The FilmArray – Data FilmArray vs DFA in 387 Pediatric NPAs Pediatric NPA samples were tested by DFA (blue) and by FilmArray (maroon).

34 82% of 387 pediatric NPA samples tested positive by FilmArray analysis

35 32% tested positive for 2 or more pathogens by FilmArray analysis

36 Future Developments Product Development (Beyond RP) Sepsis/Meningitis Panel Enteric Pathogens Panel STD Panel Biothreat Panel

37 Questions


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