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Ultra-sensitive detection of ecologically rare fish from eDNA samples based on the RPA-CRISPR/Cas12a technology
Wei, Xing-Yi1,2; Liu, Li2; Hu, Huan1,2; Jia, Huang- Jie2; Bu, Ling-Kang2; Pei, De-Sheng3
2023-09-15
摘要Environmental DNA (eDNA) research holds great promise for improving biodiversity science and conservation efforts by enabling worldwide species censuses in near real-time. Current eDNA methods face challenges in detecting low -abun-dance ecologically important species. In this study, we used isothermal recombinase polymerase amplification (RPA)-CRISPR/Cas detection to test Ctenophar-yngodon idella. RPA-CRISPR-Cas12a detected 6.0 eDNA copies/AL within 35 min. Ecologically rare species were identified in the Three Gorges Reservoir Area (TGRA) using functional distinctiveness and geographical restrictiveness, with seven fish species (9%) classified as potentially ecologically rare including three species in this investigation. RPA-CRISPR/Cas12a-FQ outperformed high-throughput sequencing (HTS) and qPCR in detecting low-abundance eDNA (AUC = 0.883**). A significant linear correlation (R-2 = 0.682**) between RPA-CRISPR/Cas12a-FQ and HTS quantification suggests its potential for predicting species abundance and enhancing eDNA-based fish biodiversity monitoring. This study highlights the value of RPA-CRISPR/Cas12a-FQ as a tool for advancing eDNA research and conservation efforts.
DOI10.1016/j.isci.2023.107519
发表期刊ISCIENCE
卷号26期号:9页码:18
通讯作者Pei, De-Sheng(peids@cqmu.edu.cn)
收录类别SCI
WOS记录号WOS:001062639500001
语种英语