Rapid field testing of field water samples for total coliform and E. coli using isothermal amplification and unique micron-sized particles for cell capture and concentration

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.