2023 Early Hearing Detection & Intervention Conference
March 5-7, 2023 • Cincinnati, OH
5/22/2018 | 9:15 AM - 9:30 AM | COMPARISON OF QUANTITATIVE PCR WITH DIGITAL DROPLET PCR FOR DETECTION AND QUANTIFICATION OF ENVIRONMENTAL DNA FROM INVASIVE ASIAN CARP | 420 B
COMPARISON OF QUANTITATIVE PCR WITH DIGITAL DROPLET PCR FOR DETECTION AND QUANTIFICATION OF ENVIRONMENTAL DNA FROM INVASIVE ASIAN CARP
Invasive Asian Carp include four species: Silver Carp, Bighead Carp, Grass Carp, and Black Carp. All four species have invaded the Mississippi River system and currently threaten to invade the Great Lakes. Environmental DNA (eDNA) is being used for early detection and monitoring of Asian Carp populations. Quantitative PCR (qPCR) assays are currently used for targeted analysis of Asian Carp eDNA because they provide both detection and an estimate of the concentration of eDNA of the target species. We tested digital droplet PCR to evaluate its potential as a replacement or complement to qPCR analysis. Potential advantages of ddPCR are less variation at low concentrations of the target sequence, absolute quantification independent of a standard curve, and resistance to PCR inhibitors in water samples. We compared the limit of detection (LOD) and limit of quantification (LOQ) of both techniques for known concentrations of synthetic DNA containing an Asian Carp target sequence. We further compared the performance of ddPCR and qPCR with environmental samples from areas invaded by Asian Carp, and finally compared their performance with environmental samples spiked with varying levels of algae, a common source of PCR inhibitors.
- Fish
- Invasive
- Monitoring
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Presenters/Authors
Catherine Richter
(), Columbia Environmental Research Center, USGS, CRichter@usgs.gov;
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