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Rapid Bacterial Detection System Fully Automated 4 minute CFU/ml response system

Rapid Bacterial Detection System Fully Automated 4 minute CFU/ml response system. SUBC Inc. Rochester MN. Principle and Sequence of Rapid Detection. 1. Platelets and cellular debris removed from platelet concentrate sample

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Rapid Bacterial Detection System Fully Automated 4 minute CFU/ml response system

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  1. Rapid Bacterial Detection SystemFully Automated 4 minute CFU/ml response system SUBC Inc. Rochester MN

  2. Principle and Sequence of Rapid Detection 1. Platelets and cellular debris removed from platelet concentrate sample 2. Present Bacteria are isolated and immobilized in isolation chamber 3. Isolation chamber rinsed with buffer, leaving isolated bacteria 4. Lyse solution in combination with localized heat provide rapid bacterial membrane lyse 5. Bacterial ATP containing lysate is mixed with bioluminescent firefly extract 6. Generated light reaction is detected by photon counter. Burst of light is correlated to CFU/ml

  3. Fluidic Analysis for Detection of Bacteria Sampling Sample Processing Detection Dilution Mixing Enrichment Chemistry Carrier Stream Waste

  4. Detector Holding coil Reactor 7 8 6 Carrier 9 5 10 4 Selection valve 1 3 2 Sample Sequential Injection Analysis for Bacterial Detection SIA Reagent Pump

  5. Platelets Platelet removal System Rinse Lyse + Heat Bacteria Isolation Chamber waste Reactor CFU Rapid Rapid Fluidic Engine Sequence Bioluminescent Reagent 3 Minute Result

  6. Firefly Luciferace Reaction ATP +D-luciferin + O2 AMP+pyrophosphate+oxyluciferin+CO2 + Light • Thequantum efficiency is very high resulting in almost one photon per ATP molecules consumed in the reaction • Intensity of the emitted light is proportional to the ATP concentration • If the luciferase level is low the intensity will be essentially constant • If the luciferase level is high the light will decay rapidly since ATP is consumed in the reaction (the initial peak light is proportional to the decay rate) • If decay rate t 1/2 = 139 minutes sensitivity range 10-15 - 10-9 • If decay rate t 1/2 = 235 minutes sensitivity range 10-14 - 10-18

  7. Prototype Device #1

  8. BPAC December 2002 Bacterial Required for Detection

  9. Bacteria verified and validated for bead isolation

  10. GloBac™

  11. Fully Automated Touch Screen Display Microprocessor Driven Internal Reagent cartridge Top loading sample port GloBac™

  12. University of Minnesota Field Evaluation Testing • Dr. McCullough • Dr. Bowman • Dr. Gundu Rao • Mary Clay: Project Manager • Shelley Pulkrabek: Blood Bank Coordinator • Debbie Cocking Johnson: Related lab support • Nancy Ward: Technical Supervisor

  13. India Study • Bangalore Rotary Blood Bank Study • Expected contamination rate of over 6% • Study size 2000 units • Expected positives greater than180 • Discussion with FDA on accepting data

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