Ultra-sensitive detection of ecologically rare fish from eDNA samples based on the RPA-CRISPR/Cas12a technology

作者全名："Wei, Xing-Yi; Liu, Li; Hu, Huan; Jia, Huang- Jie; Bu, Ling-Kang; Pei, De-Sheng"

作者地址："[Wei, Xing-Yi; Hu, Huan] Chongqing Jiaotong Univ, Sch River & Ocean Engn, Key Lab Hydraul Waterway Engn, Minist Educ, Chongqing 400074, Peoples R China; [Wei, Xing-Yi; Liu, Li; Hu, Huan; Jia, Huang- Jie; Bu, Ling-Kang] Univ Chinese Acad Sci, Chinese Acad Sci, Chongqing Inst Green & Intelligent Technol, Chongqing Sch, Chongqing 400714, Peoples R China; [Pei, De-Sheng] Chongqing Med Univ, Sch Publ Hlth, Chongqing 400016, Peoples R China"

通信作者："Pei, DS (通讯作者)，Chongqing Med Univ, Sch Publ Hlth, Chongqing 400016, Peoples R China."

来源：ISCIENCE

ESI学科分类：　

WOS号：WOS:001062639500001

JCR分区：Q1

影响因子：4.6

年份：2023

卷号：26

期号：9

开始页：　

结束页：　

文献类型：Article

关键词：　

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."

基金机构："China Sri Lanka Joint Center for Education and Research, Chinese Academy of Sciences, China"

基金资助正文："We thank the support from the Chongqing Medical University Talent Project (No. R4014 to D.S.P.), Chongqing Postdoctoral Innovation Mentor Studio (X7928 D.S.P.), China-Sri Lanka Joint Research and & nbsp;Demonstration Center for Water Technology, and China Sri Lanka Joint Center for Education and Research, Chinese Academy of Sciences, China.r Demonstration Center for Water Technology, and China Sri Lanka Joint Center for Education and Research, Chinese Academy of Sciences, China."