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Needle in a Haystack: A Droplet Digital Polymerase Chain Reaction Assay to Detect Rare Helminth Parasites Infecting
Chloe A Fouilloux1, Eric Neeno-Eckwall1, Ipsita Srinivas1
1Department of Pathobiological Sciences, University of Wisconsin-Madison, Madison, Wisconsin, USA.
Molecular Ecology Resources
|June 11, 2025
Summary
A new droplet digital PCR (ddPCR) assay accurately quantifies helminth parasite loads in wildlife and environmental samples. This sensitive method improves tracking of parasite transmission, aiding disease management in public health and agriculture.
Area of Science:
- Environmental Science
- Parasitology
- Molecular Biology
Background:
- Helminths significantly impact public health and agriculture but are understudied, especially regarding transmission dynamics in wildlife.
- Accurate quantification of helminth parasite loads in hosts and environments is crucial for understanding transmission but is challenging with traditional methods.
- Existing molecular techniques like PCR and qPCR may lack the sensitivity to detect low parasite burdens, potentially underestimating infection prevalence.
Purpose of the Study:
- To develop a sensitive and precise method for quantifying helminth parasite loads in aquatic habitats.
- To establish a tool for monitoring helminth infection dynamics across various hosts and environments.
- To address limitations in current methods for detecting and quantifying helminths in wildlife and environmental samples.
Main Methods:
- Development of a multiplexed droplet digital PCR (ddPCR) assay targeting 18S rRNA genes.
- Validation of the ddPCR assay using laboratory standards and field samples, including environmental DNA (eDNA).
- Case study utilizing the assay to quantify helminth loads in copepod intermediate hosts (Schistocephalus solidus).
Main Results:
- The ddPCR assay demonstrated high sensitivity and precision in quantifying helminth loads.
- The assay reliably detected target genes at DNA concentrations as low as 1 pg.
- The method proved effective for analyzing multi-species samples and environmental DNA (eDNA).
Conclusions:
- The developed multiplexed ddPCR assay offers a sensitive and reproducible tool for quantifying helminth parasite loads.
- This assay can enhance the monitoring of helminth infection dynamics in intermediate hosts and across spatio-temporal scales.
- The toolkit supports public health, agriculture, and conservation biology by improving the understanding and management of helminth-borne diseases.
