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FilmArray ™

FilmArray ™. Idaho Technology’s 4 th Generation Pathogen Detection System. FilmArray Beta Prototype. Idaho Technology Kirk Ririe Mark Poritz Steve Dobrowolski Team FilmArray U. Of Utah: Dr. Carrie Byington Dr. Judy Daly. FilmArray System Capabilities.

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FilmArray ™

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  1. FilmArray™ Idaho Technology’s 4th Generation Pathogen Detection System

  2. FilmArray Beta Prototype • Idaho Technology • Kirk Ririe • Mark Poritz • Steve Dobrowolski • Team FilmArray • U. Of Utah: • Dr. Carrie Byington • Dr. Judy Daly

  3. FilmArray System Capabilities • Detection of > 20 viral or bacterial agents • Clinical or environmental samples • Fully integrated automated sample processing and analysis in a closed system • Reduced self-contamination risk • Time to identification < 1 hour • Machine weight < 25lbs

  4. How Does it Work? • Nested Multiplex PCR (nmPCR) • High-sensitivity assays for large panels of biological agents. • Chemical Circuits in a Laminated Plastic Pouch • The closed system mitigates the contamination risks • inherent in nmPCR • Instrument Automation • Integrated electro-pneumatic systems • Control the movement of fluid through the pouch

  5. PCR1 Wash Dilute Cell Lysis Mag Bead Capture PCR2 FilmArray Pouch Flash

  6. PathogenGenome Outer primers, Reverse transcription 1st PCR 200bp Inner primers, 2nd PCR 90bp Nested PCR

  7. Primary RT-PCR Secondary PCR LCGreen Detection 1F 1R 2F 2R 3F 3R 4F 4R 5F 5R 6F 6R Dilute 100 fold Split between multiple singleplex reactions Nested Multiplex PCR

  8. Respiratory Virus Grant • NIH/NIAID 4 year U01 grant has funded development • A portable instrument able to perform: • Nucleic acid extraction from NPA samples • Reverse transcription • Nested multiplex PCR • High resolution DNA melting curve analysis • Automatic identification from amplification/melts • A freeze-dried reagent pouch to detect: • A panel of common respiratory viruses • SARS Coronavirus and Avian flu (H5, N1)

  9. Virus Assay Panel Many assays are required to capture diversity of viral pathogens *with automation of PCR array manufacture up to 120 second stage PCR reactions can be run

  10. Pediatric NPA Samples • Inject sample into a virus panel FilmArray pouch • 1) PCMC: FluA NPA • 2) PCMC: BocaV NPA + RSV NPA • 3) Inactivated SARS virus spiked into NPA • Hit go!

  11. FluA NPA FluA H3 FluA MA FluA N2 NL63 Pol*

  12. BocaV + RSV NPAs BoV NS-1 BoV NP-1 RSV MA FluA MA* * This was most likely miss-spotting of BoV NS-1 Primers into a neighboring FluA-MA well

  13. Inactivated SARS CoV SARS Pol SARS NG

  14. Mis-spotting and Misamplification • Manually spotting primers into the PCR array is error prone • We have automated using a Nanoplotter piezoelectric microarrayer (GeSiM) • Our current HRV and Adenovirus assays are prone to nonspecific amplification • Assay redesign and a true instrument hot start will lower this

  15. Run Summary

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