TY - JOUR
T1 - Improving reliability in environmental DNA detection surveys through enhanced quality control
AU - FURLAN, Elise
AU - GLEESON, Dianne
PY - 2017
Y1 - 2017
N2 - Species-specific environmentalDNA(eDNA) surveys are increasingly being used to infer species presence in an environment. Current inadequacies in quality control increase concern for false negatives, which can have serious ramifications for both the management of invasive species and the conservation of native species. eDNA surveys involve a multi-step process to sample, capture, extract and amplify target DNA from the environment. We outline various positive control options and show that many of the commonly used controls are capable of detecting false negatives arising during the amplification stage only. We suggest a secondary, generic primer, designed to co-amplify endogenous DNA sampled during species-specific eDNA surveys, constitutes a superior positive control to monitor method success throughout all stages ofeDNAanalysis.Wedevelop a species-specific European carp (Cyprinus carpio) assay and a generic fish assay for use as an endogenous control for eDNA surveys in Australian freshwater systems where fish are known to be abundant.We use these assays in a multiplex on eDNA samples that are simultaneously sampled, captured, extracted and amplified. This positive control allows us to distinguish method error from informative non-amplification results, improving reliability in eDNA surveys, which will ultimately lead to better informed conservation management decisions. Journal compilation.
AB - Species-specific environmentalDNA(eDNA) surveys are increasingly being used to infer species presence in an environment. Current inadequacies in quality control increase concern for false negatives, which can have serious ramifications for both the management of invasive species and the conservation of native species. eDNA surveys involve a multi-step process to sample, capture, extract and amplify target DNA from the environment. We outline various positive control options and show that many of the commonly used controls are capable of detecting false negatives arising during the amplification stage only. We suggest a secondary, generic primer, designed to co-amplify endogenous DNA sampled during species-specific eDNA surveys, constitutes a superior positive control to monitor method success throughout all stages ofeDNAanalysis.Wedevelop a species-specific European carp (Cyprinus carpio) assay and a generic fish assay for use as an endogenous control for eDNA surveys in Australian freshwater systems where fish are known to be abundant.We use these assays in a multiplex on eDNA samples that are simultaneously sampled, captured, extracted and amplified. This positive control allows us to distinguish method error from informative non-amplification results, improving reliability in eDNA surveys, which will ultimately lead to better informed conservation management decisions. Journal compilation.
KW - Detection
KW - False negative
KW - Positive control
KW - Sensitivity
KW - Type II error
UR - http://www.scopus.com/inward/record.url?scp=85011310526&partnerID=8YFLogxK
UR - http://www.mendeley.com/research/improving-reliability-environmental-dna-detection-surveys-through-enhanced-quality-control
U2 - 10.1071/MF15349
DO - 10.1071/MF15349
M3 - Article
SN - 1323-1650
VL - 68
SP - 388
EP - 395
JO - Marine and Freshwater Research
JF - Marine and Freshwater Research
IS - 2
ER -