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Related Experiment Video

Updated: Jan 13, 2026

Development and Testing of Species-specific Quantitative PCR Assays for Environmental DNA Applications
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Development and Testing of Species-specific Quantitative PCR Assays for Environmental DNA Applications

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Identifying the end user requirements for a novel, field-based environmental DNA (eDNA) forensic collection method.

Matthew Lewis1, Katie Lainé2, Kirstie Scott3

  • 1School of Pharmacy and Biomolecular Sciences, Liverpool John Moores University, Byrom Street, Liverpool, UK; Forensic Research Institute (FORRI), Liverpool John Moores University, Byrom Street, Liverpool, UK.

Science & Justice : Journal of the Forensic Science Society
|January 8, 2026
PubMed
Summary

This study evaluated new environmental DNA (eDNA) sampling methods for freshwater pearl mussel conservation. Basic and intermediate eDNA collection methods were preferred for ease of use, speed, and cost-effectiveness in forensic investigations.

Keywords:
Environmental DNA (eDNA)Forensic method developmentFreshwater pearl musselWildlife crime investigation

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Area of Science:

  • Environmental DNA (eDNA) analysis
  • Forensic science applications
  • Conservation genetics

Background:

  • Traditional surveys for freshwater pearl mussel (Margaritifera margaritifera) at crime scenes are costly, invasive, and prone to errors.
  • Environmental DNA (eDNA) offers a non-invasive alternative but requires optimized collection methods for forensic use.
  • Existing eDNA methods are not routinely implemented in wildlife crime investigations.

Purpose of the Study:

  • To identify end-user requirements for a novel eDNA sampling method for Margaritifera margaritifera.
  • To develop and compare three distinct eDNA collection approaches (basic, intermediate, advanced).
  • To assess the usability, cost-effectiveness, and data accuracy of different eDNA sampling methods for forensic applications.

Main Methods:

  • Stakeholder discussions and questionnaires identified key criteria for method development.
  • Three eDNA collection methods were developed: basic (bottles), intermediate (bags), and advanced (filtration).
  • Questionnaires assessed ease of use and risk; time and cost analyses were performed; qPCR assays evaluated data accuracy.

Main Results:

  • End-users preferred basic and intermediate eDNA collection methods due to ease of use and lower risk.
  • Basic and intermediate methods were quicker and cheaper, while the advanced method offered potential preservation benefits.
  • The advanced filtration method showed surprisingly poor eDNA detection rates, impacting data accuracy.

Conclusions:

  • Co-developed, user-friendly eDNA sampling methods are beneficial for first responders in wildlife crime investigations.
  • Basic and intermediate eDNA collection methods show promise for Margaritifera margaritifera forensic investigations.
  • Further research is needed to refine qPCR assays and validate these methods on wild populations.