Picoinjection-Enabled Multitarget Loop-Mediated Isothermal Amplification for Detection of Foodborne Pathogens

Anal Chem. 2018 Nov 20;90(22):13173-13177. doi: 10.1021/acs.analchem.8b03673. Epub 2018 Oct 24.

Abstract

In this study, we develop a method to detect multiple DNAs of foodborne pathogens by encapsulating emulsion droplets for loop-mediated isothermal amplification (LAMP). In contrast to the traditional bulk-phase LAMP, which involves a labor-intensive mixing process, with our method, different primers are automatically mixed with DNA samples and LAMP buffers after picoinjection. By directly observing and analyzing the fluorescence intensity of the resultant droplets, one can detect DNA from different pathogens, with a detection limit 500 times lower than that obtained by bulk-phase LAMP. We further demonstrate the ability to quantify bacteria concentration by detecting bacterial DNA in practical samples, showing great potential in monitoring water resources and their contamination by pathogenic bacteria.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bacteria / genetics
  • Bacteria / isolation & purification*
  • DNA, Bacterial / analysis*
  • Food Contamination / analysis*
  • Foodborne Diseases / prevention & control
  • Lab-On-A-Chip Devices
  • Limit of Detection
  • Microfluidic Analytical Techniques / methods*
  • Nucleic Acid Amplification Techniques / methods
  • Sensitivity and Specificity
  • Wastewater / analysis

Substances

  • DNA, Bacterial
  • Waste Water