(382e) A Conic Nanopore Array for Point-of-Care Microrna Profiling

We report a low-cost and turn-key Point-of-Care (POC) membrane-nanopore hybrid platform that can identify and enumerate a panel of 20-nucleotide microRNAs (miRNA) in a heterogeneous sample, with minimum pretreatment and with a dynamic range comparable to rtPCR. The approach is to integrate in a multi-stage platform existing membrane microfluidic pretreatment technologies with new solid-state conic nanopore array technologies. New nanofluidic technologies have been used to offer ultra-fine molecular isolation and enumeration capabilities: nanoparticle assembly for probe loading and non-target gating, pore surface modification to prolong translocation of short miRNA for intrapore hybridization capture and pore level ion-current Ohmic heating for precision melting of hybridization complexes to release a small number of targets. The throughput is one-thousand to one-million times higher than a single nanopore and yet the translocation time of individual miRNA is sufficiently long for hybridization. This integrated multi-stage design allows us to isolate and interrogate a minority subset of short miRNAs from a large population of molecules in a heterogeneous sample. It complements our membrane sensor design to offer a limit of detection of 100 copies and 7 decades of dynamic range.