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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

作者： Wang, Jiahong 1 ; Sun, He 2 ; Gao, Yuhan 1 ; Bu, Shengjun 2 ; Zhang, Zebin 2 ; Wang, Chen 2 ; Zhang, Hongyi 3 ; Zhang, Wenhui 1 ; Wan, Jiayu 2 ;

作者机构： 1.Jilin Agr Univ, Coll Life Sci, Changchun 130118, Peoples R China

2.Chinese Acad Agr Sci, Changchun Vet Res Inst, Changchun 130122, Peoples R China

3.Jilin Prov FAW Gen Hosp, Changchun 130011, Peoples R China

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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