Device for rapid, on-chip, sample-to-answer bacterial identification and drug susceptibility testing

PROJECT SUMMARY ABSTRACTSepsis is the leading cause of death in U Shospitals and is a medical emergency that requires immediate treatmentEvery hour that treatment is delayedsepsis related mortality increases byandof sepsis deaths can be prevented with rapid diagnosis and treatmentHenceproviding clinicians with a tool for rapid bacterial strain identification and drug susceptibility testing will enable timely and effective treatmentreduce the non specific use of broad spectrum antimicrobialsand improve clinical outcomesConventional assays for diagnosis of bloodstream infections rely on blood cultureswhich are timeconsuming and delay rapid and targeted treatment decisionsMolecular technologies that use bacterial nucleic acid amplification are faster than culture based methodsbut often require pre analytic processing which adds time and complexity to the assayAlthough some commercial assays are even more rapidtechnologies that can further expedite bacterial identification and drug screening will impact current sepsis survival ratesThe goal of this SBIR Phase I project is to demonstrate feasibility of rapid and sensitive bacterial capturestrain identification and drug susceptibility testing from whole bloodThe system is based on our novel actuating surface attached postsASAPtechnologyASAPs are an array of magnetic micro posts that are flexible in the presence of a magnetic fieldIn this projectASAP motion will expedite bacterial binding to an immobilized antibody microarray on the cartridge floor that consists of antibodies specific to several bacterial strainsThe final device will have antibodies specific to themost common bacterial strains that infect bloodwith one antibody per bacterial strainAntibodies will be arranged inrows andcolumnsEach column will contain allantibodies whose positions in thecolumns will be scrambledThe locations of all antibody spots on the microarray will be pre determinedFor this Phase I proof of concept studyantibodies to five bacterial strains will be printed on the cartridge floorFollowing bacteria capturethe chamber will be washed and the captured bacterial strain swill be immediatelyinminidentified using a fluorescent mannose lectin bindingMBLantibody that binds to MBL found on the surfaces of pathogensCaptured bacterial strain swill then be cultured in cartridgewith ASAP motion circulating the culture media to optimize bacterial growth conditions and transport newly formed bacteria to adjacent antibody spots on the microarray floorDrug solutionseach in three different concentrationsi esusceptibleintermediate and highwill be added to the bacteria and time lapse imaging will evaluate bacterial growth to determine drug susceptibilityThe significant advantage of our device is bacterial strain identification in whole blood inmin and drug screening inhourswhich arex and at leastx fasterrespectivelythan existing assays NARRATIVE A novel sample to answer microfluidic device for rapid in cartridge bacterial strain identification and drug susceptibility testing from whole blood will be developed to expedite sepsis diagnosis and treatmentand improve clinical outcomes