A novel biosensor for detecting V. parahaemolyticus based on cascade signal amplification of CRISPR/Cas14a and Exo III

文献类型： 外文期刊

第一作者： Wang, Jiahong

作者： Wang, Jiahong;Gao, Yuhan;Zhang, Wenhui;Wang, Jiahong;Sun, He;Gao, Yuhan;Bu, Shengjun;Zhang, Zebin;Wang, Chen;Wan, Jiayu;Zhang, Hongyi

作者机构：

关键词： Vibrio parahaemolyticus; Exonuclease III; CRISPR/Cas14a; Aptamer; Cyclic signal amplification

期刊名称：FOOD CONTROL （ 影响因子：5.6； 五年影响因子：5.4 ）

ISSN： 0956-7135

年卷期： 2025 年 167 卷

页码：

收录情况： SCI

摘要： Vibrio parahaemolyticus(V. parahaemolyticus)is a widespread food-borne pathogen in seafood, which poses a serious threat to human health and public safety. However, sensor platform projects that do not require complex sample processing are rare. Hence, we based on CRISPR/Cas14a and Exonuclease III (Exo III) to build a novel fluorescent biosensor that did not require nucleic acid extraction and amplification. A locked single-stranded DNA probe to a target-pathogen specific aptamer was designed to be opened as free single-stranded DNA (ssDNA) upon aptamer binding to the target pathogen. Exo III then cleaves hairpin B (HP B) from the 3 '-terminus, releasing ssDNA probe A and ssDNA C. Then, ssDNA probe A hybridizes with another HP B to start a new cycle, and ssDNA C acts as a substrate to activate Cas14a/sgRNA trans-cleavage activity. By exploiting this novel design principle, we detected V. parahaemolyticus over a wide linear range with a limit of detection (LOD) of 6 CFU/mL.

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