SBIR-STTR Award

iFyber LLC proposes to develop a coliform assay with differential detection of E. coli that will circumvent the current challenges with detecting these organisms at a level of 1 CFU/mL in less than 16 hours. The proposed assay targets a result in less than 6 hours by making use of novel microsphere reagents for capture and concentration of the target cells, and coupling these reagents with loop-mediated amplification (LAMP) of coliform and E. coli genetic markers with fluorescence detection. Microspheres will be used to concentrate the target cells by at least 1000 fold, where they will then be further enriched with growth medium, and their respective genetic reporters will be subsequently amplified through LAMP. This PhaseI effort will lay the foundation for this concept with a breadboard device, which will be refined in Phase II for commercial development.

iFyberLLC proposes to develop acoliform assay with differential detection of E.coliand total coliform that will circumvent thecurrent challenges with detectingthese organismsatalevel of1 CFU per100mLin less than 8 hours. The proposed assay targetsaresult in less than 8 hours by making use of novel microspherereagents forcaptureand concentration of thetargetcells,and couplingthesereagents with loopmediated amplification (LAMP) ofcoliform and E.coligeneticmarkers with fluorescence detection.Microspheres will be used to concentratethe targetcells byat least1000fold,wherethey will then befurtherenriched with growth medium,and their respectivegenetictargets will be subsequentlyamplified through LAMP. This PhaseIIeffort is building offasuccessful PhaseI project that showed feasibility for theconcept. PhaseII will befocused on assay optimization,kit development,and development ofaalpha prototypesystem that will bevalidated next to currentlyapproved EPA ATP methods.