Project Summary/Abstract:Invasive aspergillosis (IA) is a leading cause of morbidity and death in immunocompromised patients, with over 30million individuals at risk for IA and at least 200,000 IA cases developing worldwide each year. Despite thedevelopment of potent triazole antifungal drugs for the treatment of IA, IA-associated mortality is still 22-40%.Timely and accurate diagnosis of IA, in conjunction with prompt initiation of appropriate antifungal therapy,improves patient outcomes, but is challenging due to the shortcomings of existing diagnostics. Due to thelimitations of IA diagnostics and the potentially severe clinical consequences of not treating IA promptly, cliniciansroutinely prescribe empiric antifungal therapy to patients with suspected IA, exposing many patients who ultimatelydo not have IA to unnecessary antifungal drugs, with their toxicities and costs. A major unmet need in the care ofimmunocompromised patients is a diagnostic that can identify IA rapidly and accurately, without requiring invasiveprocedures. To address this unmet need, we propose further development of a novel microfluidic gaschromatography-differential mobility spectrometry (GC-DMS) breath analysis device for the rapid,noninvasive diagnosis of IA, based on the scientific premise that patients with IA have a unique Aspergillusvolatile sesquiterpene secondary metabolite signature in their breath. We have demonstrated this premise in aninitial proof-of-concept study and subsequent validation study in patients with suspected invasive fungal disease.Combining our expertise in engineering, diagnostic product development, and breath analysis, we have developedan integrated, fully functioning GC-DMS Microanalyzer prototype for rapid bedside breath analysis,successfully detecting these breath sesquiterpene metabolites in patients with IA using this device. The goal ofthis Fast Track SBIR proposal is further optimization and development of the GC-DMS Microanalyzer breathanalysis platform for the rapid diagnosis of IA. The objective of phase I is to optimize the preconcentration ofsesquiterpene metabolites in the Microanalyzer device, lowering its limit of detection for these analytes andevaluating its sensitivity in a set of patients with suspected invasive fungal disease. In phase II, aims are to: (1)complete Microanalyzer diagnostic device development for IA under design control, incorporating general andspecial control measures, (2) develop and validate GC-DMS peak-finding algorithms for the automatedidentification of the IA breath signature, and (3) perform comprehensive analytical validation studies for thisGC-DMS Microanalyzer breath test for IA. Upon successful completion of these objectives, the Microanalyzerbreath test for IA will be ready for evaluation in a multicenter clinical validation study for FDA 510(k) clearanceand CE mark certification. This diagnostic device will transform the care of immunocompromised patients atrisk for IA, reducing diagnostic delays, facilitating early, appropriate antifungal treatment, and improvingclinical outcomes in these patients.