2016
DOI: 10.1111/1755-0998.12619
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Abstract: A set of universal guidelines is needed to determine the limit of detection (LOD) in PCR-based analyses of low-concentration DNA. In particular, environmental DNA (eDNA) studies require sensitive and reliable methods to detect rare and cryptic species through shed genetic material in environmental samples. Current strategies for assessing detection limits of eDNA are either too stringent or subjective, possibly resulting in biased estimates of species' presence. Here, a conservative LOD analysis grounded in an… Show more

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Cited by 110 publications
(123 citation statements)
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“…Incorporating an experimentally derived threshold to remove false positives (as we did here) will also remove true detections, resulting in decreased detection probabilities. Statistical methods for estimating this threshold are also available and can be used to differentiate detections from nondetections (Hunter et al., ). Finally, model‐based methods to account for false positives also exist, and the best option to account for false positives will likely be context dependent.…”
Section: Discussionmentioning
confidence: 99%
“…Incorporating an experimentally derived threshold to remove false positives (as we did here) will also remove true detections, resulting in decreased detection probabilities. Statistical methods for estimating this threshold are also available and can be used to differentiate detections from nondetections (Hunter et al., ). Finally, model‐based methods to account for false positives also exist, and the best option to account for false positives will likely be context dependent.…”
Section: Discussionmentioning
confidence: 99%
“…Digital PCR provides sensitive and absolute DNA quantification of target DNA without requiring the use of a standard curve (Nathan, Simmons, Wegleitner, Jerde, & Mahon, ). Digital PCR has some advantages over real‐time qPCR, such as a higher tolerance to PCR inhibitors and a lower possibility of the smaller variations in estimates (Doi, Uchii, et al, ; Hunter et al, ). However, digital PCR requires expensive apparatus and has higher running costs compared with real‐time qPCR.…”
Section: Edna Detectionmentioning
confidence: 99%
“…) or optimization of low‐copy DNA detection (e.g., increased biological or technical replicates per sample or qPCR, utilization of more sensitive equipment or methods; Schultz and Lance ; Hunter et al. ) are some of the additional steps that could be tested. Thus, while the challenge of designing, testing, and validating eDNA assays may seem to be a simple list of steps to implement, it is clear that these minimum basic practices can be complex at times and worthy of careful planning and execution in every case.…”
Section: Discussionmentioning
confidence: 99%