Loop-mediated isothermal amplification (LAMP) assay-A rapid detection tool for identifying red fox (Vulpes vulpes) DNA in the carcasses of harbour porpoises (Phocoena phocoena)

PLoS One. 2017 Sep 1;12(9):e0184349. doi: 10.1371/journal.pone.0184349. eCollection 2017.

Abstract

Carcasses of wild animals are often visited by different scavengers. However, determining which scavenger caused certain types of bite marks is particularly difficult and knowledge thereof is lacking. Therefore, a loop-mediated isothermal amplification (LAMP) assay (target sequence cytochrome b) was developed to detect red fox DNA in carcasses of harbour porpoises. The MSwab™ method for direct testing without prior DNA isolation was validated. As a detection device, the portable real-time fluorometer Genie® II was used, which yields rapid results and can be used in field studies without huge laboratory equipment. In addition to in vitro evaluation and validation, a stranded and scavenged harbour porpoise carcass was successfully examined for red fox DNA residues. The developed LAMP method is a valuable diagnostic tool for confirming presumable red fox bite wounds in harbour porpoises without further DNA isolation steps.

MeSH terms

  • Animals
  • Animals, Wild
  • Cytochromes b / genetics
  • DNA / analysis*
  • Fluorometry
  • Foxes / genetics*
  • Nucleic Acid Amplification Techniques / methods*
  • Porpoises / genetics*
  • Sensitivity and Specificity

Substances

  • DNA
  • Cytochromes b

Grants and funding

The investigations were funded by the Ministry of Energy, Agriculture, Environment and Rural Areas of Schleswig-Holstein, Germany (grant number 60079084). There was no additional external funding received for this study. The funder had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.