A round-robin evaluation of the repeatability and reproducibility of environmental DNA assays for dreissenid mussels
Resource managers may be hesitant to make decisions based on environmental (e)DNA results alone since eDNA is an indirect method of species detection. One way to reduce the uncertainty of eDNA is to identify laboratory‐based protocols that ensure repeatable and reproducible results. We conducted a double‐blind round‐robin analysis of probe‐based assays for DNA of dreissenid (Dreissena spp.) mussels, which are prolific aquatic invaders that can cause significant economic and ecological impacts. DNA extract from water samples spiked with known amounts of dreissenid DNA and from water samples collected from waters with and without dreissenids were analyzed by four independent research laboratories. We used results to calculate detection repeatability within laboratories and assays, detection reproducibility among laboratories and assays, and estimated dreissenid DNA copy number precision and accuracy. Laboratory and assay repeatability and reproducibility of detection results were high, 91% and 92%, respectively. The estimated copy numbers were neither precise nor accurate for samples spiked with <773 gene copies. These results suggest that eDNA surveillance of dreissenid mussels, using the protocols evaluated herein, can generate reliable detection data for decision‐making. However, managers should be cautious about using the quantitative information often associated with eDNA detections, especially when DNA is at lower abundance. Our results provide strong support that eDNA has the potential to provide repeatable and reproducible evidence under varying laboratory conditions and for different sample water chemistries. This is reassuring since the demand for eDNA surveillance is widespread and number of laboratories that process eDNA samples is growing steadily.
Citation Information
| Publication Year | 2020 |
|---|---|
| Title | A round-robin evaluation of the repeatability and reproducibility of environmental DNA assays for dreissenid mussels |
| DOI | 10.1002/edn3.68 |
| Authors | Adam J. Sepulveda, Patrick R. Hutchins, Craig Jackson, Carl Ostberg, Matthew Laramie, Jon Amberg, Timothy Counihan, Andrew B. Hoegh, David Pilliod |
| Publication Type | Article |
| Publication Subtype | Journal Article |
| Series Title | Environmental DNA |
| Index ID | 70208025 |
| Record Source | USGS Publications Warehouse |
| USGS Organization | Northern Rocky Mountain Science Center |
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Adam Sepulveda, Ph.D.
Research Zoologist
Patrick Hutchins
Biologist
Carl Ostberg, Ph.D.
Research Fish Biologist
Jon Amberg, PhD
Center Director
Timothy Counihan
Research Fish Biologist
David S Pilliod
Supervisory Research Ecologist
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PCR results from dreissenid mussel round robin assay analyses, 2018-2019
Real-time PCR results of a round robin evaluation of 5 assays that target dreissenid mussel DNA. Water samples collected from waters with and without dreissenid mussels were analyzed using these five assays in four USGS laboratories. Samples from waters without dreissenid mussels were spiked with known amounts of dreissend DNA. - Connect
Adam Sepulveda, Ph.D.
Research ZoologistEmailPhonePatrick Hutchins
BiologistEmailCarl Ostberg, Ph.D.
Research Fish BiologistEmailPhoneJon Amberg, PhD
Center DirectorEmailPhoneTimothy Counihan
Research Fish BiologistEmailPhoneDavid S Pilliod
Supervisory Research EcologistEmailPhone