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Advances in Early Detection and Monitoring of Sepsis: Cellular Diagnostics with Neutrophil CD64 Bruce H. Davis, M.D. Trillium Diagnostics, LLC Brewer, Maine U.S.A. www.trilliumdx.com

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Severe Sepsis: Comparison With Other Diseases Crit Care Med. 29:1303, 2001. National Center for Health Statistics, 2001. American Cancer Society, 2001. *American Heart Association. 2000. Angus DC et al. Crit Care Med. 29:1303, 2001. AIDS*Colon Breast Cancer CHF Severe Sepsis Cases/100,000 Incidence of Severe SepsisMortality of Severe Sepsis AIDS* Severe Sepsis AMI Breast Cancer Estimated annual 12 million cases with suspected Sepsis in North America, UK, and Europe 18 million cases annually worldwide with 1,400 deaths per day Healthcare cost of sepsis ~$16 billion/year

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Infection: Inflammatory response

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Laboratory Indicators of Clinical Acute Inflammation Response to Infection/Sepsis Standard of Care Diagnostic Assays of Infection/Sepsis Leukocyte Counts (neutrophilia) - CBC absolute counts band counts or left shift (immaturity index) Cultures for suspected infection Sedimentation Rate C-Reactive Protein (CRP) New Generation Assays of Infection/Sepsis Granulocyte or PMN CD64 (Leuko64, Trillium Diagnostics) Cytokine and receptor levels (intracellular or plasma, TNF-a, IL-6 (Septest), IL-12, CD14, CD16, etc.) Procalcitonin plasma levels (Brahms) Endotoxin Activity Assay (Spectral Diagnostics) TREM-1 (Triggering Receptor Expressed on Myeloid cells) LightCycler SeptiFast Test (Roche) Triage Sepsis Panel (CRP, MIP-3, NGal Biosite)

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Myeloid Antigen Maturation Sequence From Wood and Borowitz (2006), Henrys Laboratory Medicine

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Scientific Basis for Quantitative PMN CD64 as an Improved Diagnostic Test of Infection/Sepsis PMN CD64 expression is negligible in the healthy state (

PMN CD64 Index reported with values of >1.50 indicative of infection

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Leuko64 Software: Summary Report

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PMN CD64 as a Sepsis Marker in Neonates Fjaertoft et al Pediatr Res 45:871-76, 1999 Similar reports by: Layseca-Espinosa et al. Pediatr Allergy Immunol, 2002 13(5): 319-27, Ng et al. Pediatr Res, 2002. 51(3): 296-303, and Ng et al. Pediatr Res, 2004; 56(5): 796-803

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Neonatal Study: PMN CD64 Index correlates best with C- Reactive Protein and presence of sepsis Y=1.0404X 1.068 r = 0.6732

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Inflammatory Markers in Neonates

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Characteristics Healthy Controls (n=55) suspected DIC (n=97) P value IL-6 (pg/ml) 4.7 12.6 115.6 134.7 0.002 IL-10 (pg/ml) 7.8 10.5 29.0 55.4 29.0 55.4 0.002 Neutrophil CD64 1117 221 1117 221 4071 3707 4071 3707