DESCRIPTION (provided by applicant): The key objective of this proposed product development effort is to optimize the treatment of infections and reduce adverse events associated with inappropriate antibiotic use according to each NICU's locally established antibiogram with a fully automated, evidence-based rapid PID/AST system. The ultimate goal of the proposed neonatal sepsis CentriCapillary system after successful completion of this SBIR fast track is to initially address analysis needs in NICU settings then in the entire blood culture market (currently estimated to total more than $3 billion in the United States alone). Neonates are not small adults. It is not practical to design a diagnostics system using 10 mL blood culture bottles for adults and only load 1 mL of blood sample for neonates. Undoubtedly, there is a risk of missing the target pathogen for extremely low-level bacteremia (e.g., below 1 CFU/mL) using small blood sample volume, but it is not practical or even possible to obtain more than 1 mL of blood routinely from neonates in the NICU for PID and AST lab tests. Currently, there is no commercial system or product development effort from major diagnostics companies to address neonatal sepsis screening specifically using smaller blood volume. We developed, demonstrated and published an innovative molecular-based genotypic-phenotypic-hybrid approach for multiplexed bacterial PID and AST profiling with 100% clinical sensitivity, 96% clinical specificity, 98% minimum inhibitory concentration (MIC) and 97% categorical agreement in our most recent ongoing clinical feasibility study on 73 raw clinical urine samples. In this fas track SBIR project, we will leverage our expertise in microfluidics, electrochemical biosensors, and clinical microbiology to develop an integrated CentriCapillary system. While our molecular analysis technology has been validated with multidrugresistant pathogens (MDR) that cause UTI, which is the most common bacterial infection, it is anticipated that the incorporation of the lysis-centrifugation and dynamic hybridization will have applicability for rapid neonatal sepsis PID/AST and other emerging infectious diseases. PHASE I STUDY (YEAR 1) Specific Aim 1: Transition the current PID/AST platform technology from urinary tract infection (UTI) to neonatal sepsis- Hypothesis: Lysis centrifugation can address the change of matrix effect from raw urine to whole blood samples and the issue of low abundant pathogen for neonatal sepsis by using a CentriCapillary cartridge for blood- pelleting assisted rapid blood culture on various low blood volumes from neonates (10 μL - 2 mL). Specific Aim 2: Demonstrate the feasibility of the electrochemical-based molecular analysis CentriCapillary cartridge - Hypothesis: Microfluidic cartridge-based blood pelleting can capture and recover all common pathogens on the YNHH NICU's antibiogram for rapid neonatal sepsis PID and AST. PHASE II STUDY (YEARS 2 AND 3). Specific Aim 3: Develop a dual-mode electrochemical-based dynamic hybridization analysis algorithm to expand the PID/AST panel - Hypothesis: Dynamic hybridization analysis can be utilized to expand the species-specific identification of common and emerging pathogens for neonatal sepsis. Specific Aim 4: Prototype, validate and manufacture the CentriCapillary cartridge (COGS<$15) and system (COGS<$25k) - Hypothesis: Ultracentrifugation (up to 50,000 g, gravitational force) can be incorporated into a multiplexed fluidic cartridge for a fully automated neonatal sepsis PID/AST from whole blood samples in 6 hours. Specific Aim 5: Clinically validate the rapid neonatal PID/AST CentriCapillary system according to CLSI guidelines - Hypothesis: Blood samples spiked with ATCC strain bacteria used in the analytical validation studies represent critical matrix characteristics of fresh whole blood samples from neonates The development and validation of neonatal sepsis will adhere to the ISO 13485 standards, FDA/CDC/CLSI guidelines on AST and federal regulation 21 CFR 866.1645. The ultimate goal of this study is to reduce adverse events associated with inappropriate antibiotic use in the NICU settings.

PUBLIC HEALTH RELEVANCE: There are 4.1M births annually in the United States with 1 in 8 births classified as premature. Premature infants are at 30% greater risk of major medical issues, and the financial impact is $26.2B annually, or $52K per infant. There are more than 20,000 NICU beds in the United States, but there are no technologies or products available specifically to address the low blood volume drawn from neonates. The ultimate goal of the proposed rapid neonatal sepsis screening system after successful completion of this clinical feasibility study is to initially address analysis needs in NICU settings then in the entire blood culture market (currently estimated to total more than $3 billion in the United States alone) with GeneFluidics' current molecular analysis pathogen identification (PID) and antimicrobial susceptibility testing (AST) platform.