The goal of this project is to develop a molecular triage tool with enhanced performance characteristicswhich will improve the diagnosis and management of bacterial sepsis in acute care settings. A quantitativePCR-based assay has been developed which permits broad-spectrum bacterial detection coupled withspecific pathogen identification. Prior work suggested favorable detection accuracy for biothreat (BT) or non-BT related organisms in inert sample matrices.
Specific aims for this 2-years proposal include 1) to furtheroptimize the assay and perform advanced analytical validation for detection of Eubacteriales using mock-uphuman blood specimens, 2) to evaluate the performance characteristics of the assay for the diagncsis ofbacterial sepsis and clinical outcome prediction in the ED setting. Significant inroads have been made inyear 1 which included) optimized sample preparation protocol with highly sensitive (1 cfu/ml) detection limitfor eubacteria in whole blood; 2) highly specific early Gram-type classification of detected pathoger; 3)alternative post-PCR amplicon analysis methods based on molecular beacon hybridization chemistry andhigh resolution melting curve analysis for high throughput pathogen identification; and 4) IRB approval forprocuring clinical samples from ICU (in addition to ED) patients with suspected sepsis. Formal coursework inyear 1 include GMP training, Clinical Development of Drugs and Biologies, Research in Ethics and Integrity.Prospective clinical validation study of the optimized assay protocol will be the main focus for year 2 withcontinued supplemental coursework for career development. Toward the end of this award, I will beeminently trained to undertake independent translational research and pursue a productive research careerin advancing diagnostic technologies for rapid detection of biothreat and emerging infectious diseases.
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