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Point-of-Care Pulmonary Colonization Detection Protocol based upon an Ultra-Fast Gas Chromatograph

Point-of-Care Pulmonary Colonization Detection Protocol based upon an Ultra-Fast Gas Chromatograph. Technology Description Screening for chemical biomarkers In-vitro Culture Results POC Analysis Summary . Edward J. Staples, Ph.D., Q&A Solutions, Inc. and

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Point-of-Care Pulmonary Colonization Detection Protocol based upon an Ultra-Fast Gas Chromatograph

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  1. Point-of-Care Pulmonary Colonization Detection Protocol based upon an Ultra-Fast Gas Chromatograph • Technology Description • Screening for chemical biomarkers • In-vitro Culture Results • POC Analysis • Summary Edward J. Staples, Ph.D., Q&A Solutions, Inc. and Chris Landon, M.D., Director of Pediatrics, Ventura County Medical Center

  2. Cystic Fibrosis • Cystic fibrosis is one of the most common fatal genetic disorders in the United States, affecting about 30,000 individuals. • As a result of a genetic disorder, there is the presence of a thick mucus secretion which clogs the bronchial tubes in the lungs. Mucus blocks the airway passages and results in a predisposition toward chronic bacterial infections. • The most common bacterium to infect the CF lung is Pseudomonas aeruginosa, a gram-negative microorganism with a propensity to live in warm, wet environments. • Objective : To improve diagnosis speed and more effective antibiotics to improve lung function in CF patients

  3. Approach: Odors associated with Bacteria • Some bacteria have a very distinctive odor. Pseudomonas aeurginosa has a "grape-like" fruity odor. • Many think that Proteus mirabilis smells like chocolate cake. • sulphur-like smell may well be Salmonella typhi. • Many anaerobes, bacteria that grow without oxygen, have very distinctive odors as well.

  4. Point-of-Care Breath Analysis Screening for Cystic Fibrosis in Children Breath Sample Collection Chemical Biomarker Analysis with GC/SAW (zNose®)

  5. Point-of-Care Analysis using GC/SAW ( Ultra-Fast and Portable Gas Chromatography ) Portable (5 hour battery life) Disposable He =>350 runs 10-20 Second Chromatograms Universal Specificity C4-C24 Electronically Variable Sensitivity Blue-tooth connectivity Disposable He tank And battery GC/SAW Sensor Head

  6. Chemical Odor Libraries Using Kovats Indices Index Odor Chemical Name 8 Seconds

  7. Protocol: Breath Sample Collection aided by External Thairapy® Vest

  8. POC Analysis Protocol Pre-concentrator (Tenax Absorber) 100:1 enrichment  femto-gram sensitivity

  9. Calibration (Tuning) with n-alkane standard(e.g. 125 ppm in ethanol) “magic perfume” …Allows electronic nose researchers to collaborate and compare their findings untuned C6 ….C22 20 Seconds Virtual Chemical Sensors tuned Indices 400-3000

  10. Biomarkers from P. Aeruginosa Zechman etal 1985 1 butanol Isopentanol Toluene 1-undecene 2-butanone 2-heptanone 2-nonanone 2-undecanone 2-aminoacetophenone Labows etal 1980 2-nonanone 2-undecanone 2-aminoacetophenone Dimethyldisulfide Dimethyltrisulfide Butanol 2-butanone 1-undecene Isopentanol

  11. 2-nonanone Standard Response10 sec sample 60 deg detector Amount Index Graphic regions define virtual chemical sensor arrays

  12. TSB Culture medium (TSB) Aroma Responsemcarthy pa no 3 30 sec 20 deg det 165 valve, 200 inlet, 10ps-2a1b

  13. Virtual Chemical Sensor Array based upon Indexed compounds in a PA Culture Aroma TSB

  14. A. Duarte No. 1 C. Lazenby No. 3 A. Duarte No. 2 A. Duarte No. 3 M Parez No. 3 C. lazenby No. 1 C. Lazenby No. 2 Cultures from CF patients show repeatable PA Characteristic pattern (Grape odor)

  15. Comparing direct breath sample (black) with sputum culture headspace(Patient: M. Perez) Hospital room Background (citronellal) Breath Ketones …sputum vs culture vs breath sample

  16. Virtual Chemical Sensor Arrays + Pattern Recognition Amino acetophenone nonanone

  17. Cyclohexane, 1-methyl-4-(1-methylethyl)-, trans- 980-997 (SE-30) Benzene, pentamethyl- 1290 (HP-5) Cyclohexane, 1-methyl-4-(1-methylethyl)-, cis- 990-1000 Naphthalene 1180-1205 (db5) Methylcyclododecane 1380 (squalane) 3-Heptanone, 5-methyl- 939-954 (dB5) 923 (HP-1) 3-Heptanone, 2-methyl- 920 (HP-1) Cyclohexane, 1,4-dimethyl-, trans- 776-785 (db5) 3-Heptanone, 4-methyl- Cyclohexane, 1,4-dimethyl-, cis- 805 3-Heptanone, 6-methyl- Heptane 700 Biomarkers from Tuberculosis

  18. Research in Progress….TB Biomarker Virtual Sensor Array TB Virtual Sensor Array

  19. Summary • Ultra-fast chromatography provides a portable and sensitive POC technology for odor analysis • Standards empower collaboration and comparison of researcher findings • Diagnosis of Pulmonary infections such as CF and TB can be detected and quantitatively monitored using UF/GC • Specific Virtual Chemical Sensor Arrays using biomarker chemicals enable more accurate recognition of pulmonary infections • Breath, Sputum, and Culture Analysis can lead to early detection and diagnosis of pulmonary infections

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