Optical Imaging of Paramagnetic Bead-DNA Aggregation Inhibition Allows for Low Copy Number Detection of Infectious Pathogens

PLoS One. 2015 Jun 11;10(6):e0129830. doi: 10.1371/journal.pone.0129830. eCollection 2015.

Abstract

DNA-paramagnetic silica bead aggregation in a rotating magnetic field facilitates the quantification of DNA with femtogram sensitivity, but yields no sequence-specific information. Here we provide an original description of aggregation inhibition for the detection of DNA and RNA in a sequence-specific manner following loop-mediated isothermal amplification (LAMP). The fragments generated via LAMP fail to induce chaotrope-mediated bead aggregation; however, due to their ability to passivate the bead surface, they effectively inhibit bead aggregation by longer 'trigger' DNA. We demonstrate the utility of aggregation inhibition as a method for the detection of bacterial and viral pathogens with sensitivity that approaches single copies of the target. We successfully use this methodology for the detection of notable food-borne pathogens Escherichia coli O157:H7 and Salmonella enterica, as well as Rift Valley fever virus, a weaponizable virus of national security concern. We also show the concentration dependence of aggregation inhibition, suggesting the potential for quantification of target nucleic acid in clinical and environmental samples. Lastly, we demonstrate the ability to rapidly detect infectious pathogens by utilizing a cell phone and custom-written application (App), making this novel detection modality fully portable for point-of-care use.

MeSH terms

  • Animals
  • Cell Phone
  • DNA / blood*
  • DNA / chemistry*
  • DNA Primers / chemistry
  • Escherichia coli Infections / diagnosis*
  • Escherichia coli Infections / microbiology
  • Escherichia coli O157 / genetics
  • Escherichia coli O157 / isolation & purification
  • Humans
  • Magnetics
  • Nucleic Acid Amplification Techniques / methods*
  • Optical Imaging / methods*
  • Point-of-Care Systems
  • Polymerase Chain Reaction
  • Rift Valley Fever / diagnosis*
  • Rift Valley Fever / virology
  • Rift Valley fever virus / genetics
  • Rift Valley fever virus / isolation & purification
  • Salmonella enterica / genetics*
  • Salmonella enterica / isolation & purification
  • Silicon Dioxide / chemistry

Substances

  • DNA Primers
  • Silicon Dioxide
  • DNA

Grants and funding

The authors have no support or funding to report.