文献类型：期刊文献

英文题名：The development of RPA-CRISPR/Cas12a assay for the detection of Pseudomonas plecoglossicida

作者：Li, Yiming[1];Du, Haoran[1];Chen, Jiaxi[1];Xiao, Jingfan[1,2];Ma, Yue[1,2];Zhang, Yibei[1,2];Wang, Qiyao[1,2]

机构：[1]East China Univ Sci & Technol, State Key Lab Bioreactor Engn, Lab Aquat Anim Dis, Shanghai 200237, Peoples R China;[2]Shanghai Engn Res Ctr Maricultured Anim Vaccines, Shanghai 200237, Peoples R China

年份：2026

卷号：373

外文期刊名：FEMS MICROBIOLOGY LETTERS

收录：;WOS:【SCI-EXPANDED(收录号:WOS:001803695000001)】；

基金：This research was funded by the Shanghai Agricultural Science and Technology Innovation Project (T2023328) and China Agriculture Research System of MOF and MARA (CARS-46).

语种：英文

外文关键词：Pseudomonas plecoglossicida; recombinase polymerase amplification (RPA); CRISPR/Cas12a; rapid detection; aquaculture

摘要：Pseudomonas plecoglossicida is the etiological agent of visceral white spot disease, which induces significant mortality in economically important fish such as the large yellow croaker. In this study, we integrated recombinase polymerase amplification (RPA) with CRISPR/Cas12a-mediated detection to establish a fluorescence-based assay for rapid identification of P. plecoglossicida. The complete single-tube, two-stage RPA-CRISPR/Cas12a workflow can be performed within similar to 45 min. Using purified genomic DNA, the assay achieved an analytical detection limit of 1.65 copies mu l(-1) and showed no cross-reactivity with several other common fish pathogens. Its applicability was further evaluated using crude DNA extracts from spleen, liver, and kidney tissues of experimentally infected large yellow croakers. Overall, with its rapid turnaround, minimal equipment requirement, and high sensitivity, the RPA-CRISPR/Cas12a assay represents a promising diagnostic tool for rapid detection of P. plecoglossicida, thereby helping to control the spread of infection.