The spread of antibiotic-resistant bacteria poses a global threat to public health. Conventional bacterial detection and identification methods often require pre-enrichment and/or sample preprocessing and purification steps that can prolong diagnosis by days. We have developed methods to specifically capture and detect antibiotic-resistant bacteria directly from biological specimens with no prior culture and minimal processing steps using a microfluidic device and nanoparticle-mediated bacterial capture. Different approaches for bacterial capture are employed based on the origin of resistance in target strains. If extracellular markers confer resistance (e.g. membrane-bound protein penicillin binding protein 2A in methicillin-resistant S. aureus), magnetic nanoparticles functionalized with antibodies are used to recognize whole bacteria. If intracellular markers confer resistance (e.g. Klebsiella pneumoniae carbapenemase in cabapnem-resistant E. coli), magnetic nanoparticles functionalized with single stranded nucleic acid sequences complementary mRNAs that confer antibiotic resistance are employed to capture permeabilized bacteria; this approach can also be employed to detect mRNA coding for extracellular markers. Bacteria identification is then achieved via electrochemistry or fluorescence microscopy, allowing for sensitive detection of target bacteria in a few hours.

Detection of antibiotic-resistant bacteria using nanoparticle-mediated microfluidic capture

2019 Fall National Meeting

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