Droplet microfluidics for amplification-free genetic detection of single cells

Lab Chip. 2012 Sep 21;12(18):3341-7. doi: 10.1039/c2lc40537g. Epub 2012 Jul 30.

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.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Escherichia coli / isolation & purification*
  • Escherichia coli / metabolism
  • Fluorescence Resonance Energy Transfer
  • Microfluidics / instrumentation
  • Microfluidics / methods*
  • Nucleic Acid Hybridization
  • Peptide Nucleic Acids / chemistry
  • Peptide Nucleic Acids / metabolism
  • RNA, Ribosomal, 16S / metabolism

Substances

  • Peptide Nucleic Acids
  • RNA, Ribosomal, 16S