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. 2022 Jan 25;8:819681. doi: 10.3389/fvets.2021.819681

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Copyright © 2022 Li, Lin, Feng, Lin, Guan, Chen, Liang, Wang, Li, You and Chen.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Detection of AHPND in field samples using the new LFS-based RPA-CRISPR/Cas12a assay compared to real-time qPCR. (A) Schematic diagram of the assay process using field samples. Specificity of the (B) fluorescence-based and (C) LFS-based RPA-CRISPR/Cas12a assays. No-crRNA means the reaction without crRNA. AHPND-crRNA means crRNA targeted to AHPND pathogen, EHP-crRNA and WSSV-crRNA targeted to EHP and WSSV pathogen, respectively. (D) Detection Ct value of qPCR in field samples. Field samples below the blue dashed line indicate the positive results. (E) Agreement between LFS-based RPA-CRISPR/Cas12a and qPCR assays for detecting AHPND in 60 field samples of shrimp. Error bars represent the standard deviation from three independent experiments.