PROJECT SUMMARY
Sepsis, commonly caused by bloodstream infections (BSI), is a rapidly progressive and life-threatening disease.
Unfortunately, prolonged delay in microbiological diagnosis increases patient mortality, promotes the misuse of
antibiotics, and consequently, the evolution of antibiotic-resistant pathogens. Herein, we aim to deliver an
amplification-free, microfluidic system for pathogen detection, identification (ID), and antimicrobial susceptibility
testing (AST) directly from whole blood. To achieve our goal, we propose a platform based on microfluidic-
assisted microscopy to sort, trap, detect, and monitor pathogens at single cell resolution. For pathogen ID, we
will adopt a multispectral barcoding scheme to differentially label molecular probes for direct multiplex ribosomal
RNA (rRNA) detection to classify and speciate pathogens, along with a nanotube assisted microwave
electroporation (NAME) technique to efficiently deliver the probes intracellularly for amplification-free single
microbe detection. Positive pathogen ID will guide quantitative multimodal phenotypic AST (mPhAST), in which
we will monitor early changes in microbial growth kinetics with cytological measures of viability in response to
relevant antibiotic conditions at the single cell level to determine susceptibility/resistance with improved speed
and reliability. Combined with upstream whole blood pre-processing for pathogen isolation and concentration
followed by ID then AST, we aim to deliver sample to answer within 90 min for BSI triage and as early as 30
minutes more for antibiotic minimum inhibitory concentration (MIC) determination. We have assembled a superb
team of multi-disciplinary investigators and industry-leading advisors with complementary expertise and a strong
track record of collaboration. We propose the following aims: 1) to develop a rapid BSI triage protocol for broad
pathogen detection, classification, and ID; 2) to develop a quantitative mPhAST; 3) to develop an integrated ID-
mPhAST platform; 4) to perform analytical and clinical validation of our ID-mPhAST platform. Our short-term
goal is to obtain the necessary preliminary data to plan for product development and commercialization, with the
long-term goal of translating our diagnostic platform to reduce sepsis-related morbidity and mortality.
