A rapid multiplex PCR assay for presumptive species identification of rhinoceros horns and its implementation in Vietnam

PLoS One. 2018 Jun 14;13(6):e0198565. doi: 10.1371/journal.pone.0198565. eCollection 2018.

Abstract

Rhinoceros (rhinos) have suffered a dramatic increase in poaching over the past decade due to the growing demand for rhino horn products in Asia. One way to reverse this trend is to enhance enforcement and intelligence gathering tools used for species identification of horns, in particular making them fast, inexpensive and accurate. Traditionally, species identification tests are based on DNA sequence data, which, depending on laboratory resources, can be either time or cost prohibitive. This study presents a rapid rhino species identification test, utilizing species-specific primers within the cytochrome b gene multiplexed in a single reaction, with a presumptive species identification based on the length of the resultant amplicon. This multiplex PCR assay can provide a presumptive species identification result in less than 24 hours. Sequence-based definitive testing can be conducted if/when required (e.g. court purposes). This work also presents an actual casework scenario in which the presumptive test was successfully utlitised, in concert with sequence-based definitive testing. The test was carried out on seized suspected rhino horns tested at the Institute of Ecology and Biological Resources, the CITES mandated laboratory in Vietnam, a country that is known to be a major source of demand for rhino horns. This test represents the basis for which future 'rapid species identification tests' can be trialed.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Buffaloes
  • Conservation of Natural Resources
  • Cytochromes b / genetics
  • Horns*
  • Humans
  • Molecular Typing / methods*
  • Perissodactyla / genetics*
  • Polymerase Chain Reaction / methods*
  • Sequence Analysis, DNA / methods
  • Species Specificity
  • Vietnam

Substances

  • Cytochromes b

Grants and funding

The authors would like to thank the Australian Museum Foundation (https://australianmuseum.net.au/foundation) (RNJ), particularly Chris Grubb and Peter Warne, the University of Sydney (https://sydney.edu.au), TRACE Wildlife Forensic Network (https://www.tracenetwork.org) (RM), and the U.S. Agency for International Development, under the Wildlife Trafficking, Response, Assessment, and Priority Setting (Wildlife TRAPS) Project, award number AID-AID-EGEE-IO-13-00002 (see Acknowledgements section) for funding.