title: Droplet microfluidics for amplification-free genetic detection of single cells.
abstract: In this article we present a novel droplet microfluidic chip enabling amplification-free detection of single pathogenic cells. The device streamlines multiple functionalities to carry out sample digitization, cell lysis, probe-target hybridization for subsequent fluorescent detection. A peptide nucleic acid fluorescence resonance energy transfer probe (PNA beacon) is used to detect 16S rRNA present in pathogenic cells. Initially the sensitivity and quantification abilities of the platform are tested using a synthetic target mimicking the actual expression level of 16S rRNA in single cells. The capability of the device to perform "sample-to-answer" pathogen detection of single cells is demonstrated using E. coli as a model pathogen.
authors: Department of Biomedical Engineering, Johns Hopkins University, Baltimore, USA.
authors: Tushar D
authors: TD
authors: Rane
authors: Helena C
authors: HC
authors: Zec
authors: Chris
authors: C
authors: Puleo
authors: Abraham P
authors: AP
authors: Lee
authors: Tza-Huei
authors: TH
authors: Wang
mesh_terms: D004926:Escherichia coli / Q000302:isolation & purification* / Q000378:metabolism; D031541:Fluorescence Resonance Energy Transfer; D044085:Microfluidics / Q000295:instrumentation / Q000379:methods*; D009693:Nucleic Acid Hybridization; D020135:Peptide Nucleic Acids / Q000737:chemistry / Q000378:metabolism; D012336:RNA, Ribosomal, 16S / Q000378:metabolism
chemical_list: D020135:Peptide Nucleic Acids; D012336:RNA, Ribosomal, 16S