Reporting the limits of detection and quantification for environmental DNA assays

dc.contributor.authorKlymus, Katy E.
dc.contributor.authorMerkes, Christopher M.
dc.contributor.authorAllison, Michael J.
dc.contributor.authorGoldberg, Caren S.
dc.contributor.authorHelbing, Caren C.
dc.contributor.authorHunter, Margaret E.
dc.contributor.authorJackson, Craig A.
dc.contributor.authorLance, Richard F.
dc.contributor.authorMangan, Anna M.
dc.contributor.authorMonroe, Emy M.
dc.contributor.authorPiaggio, Antoinette J.
dc.contributor.authorStokdyk, Joel P.
dc.contributor.authorWilson, Chris C.
dc.contributor.authorRichter, Catherine A.
dc.date.accessioned2020-07-22T20:23:27Z
dc.date.available2020-07-22T20:23:27Z
dc.date.copyright2019en_US
dc.date.issued2019
dc.description.abstractBackground Environmental DNA (eDNA) analysis is increasingly being used to detect the presence and relative abundance of rare species, especially invasive or imperiled aquatic species. The rapid progress in the eDNA field has resulted in numerous studies impacting conservation and management actions. However, standardization of eDNA methods and reporting across the field is yet to be fully established, with one area being the calculation and interpretation of assay limit of detection (LOD) and limit of quantification (LOQ). Aims Here, we propose establishing consistent methods for determining and reporting of LOD and LOQ for single‐species quantitative PCR (qPCR) eDNA studies. Materials & Methods/ Results We utilize datasets from multiple cooperating laboratories to demonstrate both a discrete threshold approach and a curve‐fitting modeling approach for determining LODs and LOQs for eDNA qPCR assays. We also provide details of an R script developed and applied for the modeling method. Discussion/Conclusions Ultimately, standardization of how LOD and LOQ are determined, interpreted, and reported for eDNA assays will allow for more informed interpretation of assay results, more meaningful interlaboratory comparisons of experiments, and enhanced capacity for assessing the relative technical quality and performance of different eDNA qPCR assays.en_US
dc.description.reviewstatusRevieweden_US
dc.description.scholarlevelFacultyen_US
dc.description.sponsorshipThis project was funded in part by USGS's Ecosystems Mission Area, Invasive Species Program, the intramural research program of the U.S. Department of Agriculture, Animal and Plant Health Inspection Service, Wildlife Services, National Wildlife Research Center and an Ignite Grant #IGN071717 from Innovate BC. We thank Nick Akins, Lauren Bergman, Leah Botkin, Erica Mize, Nikolas Grueneis, Xin Guan, Danyang (Sandy) Shen, Nathan Thompson, Kyle Von Ruden, Alvi Wadud, and Kristyne Wozney for technical support.en_US
dc.identifier.citationKlymus, K. E., Merkes, C. M., Allison, M. J., Goldberg, C. S., Helbing, C. C., Hunter, M. E., Jackson, C. A., Lance, R. F., Mangan, A. M., Monroe, E. M., Piaggio, A. J., Stokdyk, J. P., Wilson, C. C., & Richter, C. A. (2019). Reporting the limits of detection and quantification for environmental DNA assays. Environmental DNA, 2(3), 271-282. https://doi.org/10.1002/edn3.29.en_US
dc.identifier.urihttps://doi.org/10.1002/edn3.29
dc.identifier.urihttp://hdl.handle.net/1828/11956
dc.language.isoenen_US
dc.publisherEnvironmental DNAen_US
dc.subjectassay optimization
dc.subjecteDNA
dc.subjectqPCR
dc.subjectstandardization
dc.subject.departmentDepartment of Biochemistry and Microbiology
dc.titleReporting the limits of detection and quantification for environmental DNA assaysen_US
dc.typeArticleen_US

Files

Original bundle
Now showing 1 - 1 of 1
Loading...
Thumbnail Image
Name:
Klymus_KatyE_EnvironDNA_2019.pdf
Size:
1.73 MB
Format:
Adobe Portable Document Format
Description:
License bundle
Now showing 1 - 1 of 1
No Thumbnail Available
Name:
license.txt
Size:
1.71 KB
Format:
Item-specific license agreed upon to submission
Description: