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Electrochemical biosensor for detecting pathogenic bacteria based on a hybridization chain reaction and CRISPR-Cas12a

文献类型： 外文期刊

作者： Liu, Xiu 1 ; Bu, Shengjun 2 ; Feng, Jingqi 1 ; Wei, Hongguo 1 ; Wang, Ze 2 ; Li, Xue 2 ; Zhou, Hongyu 2 ; He, Xiuxia 1 ; Wan 1 ;

作者机构： 1.Changchun Univ Sci & Technol, Sch Life Sci & Technol, Changchun 130022, Jilin, Peoples R China

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

关键词： Detect; DNA hairpin; Hybridization chain reaction; Lba Cas12a; S; typhimurium

期刊名称：ANALYTICAL AND BIOANALYTICAL CHEMISTRY （ 影响因子：4.157； 五年影响因子：3.868 ）

ISSN： 1618-2642

年卷期：

页码：

收录情况： SCI

摘要： In this study, Lba Cas12a (Cpf1) as one of the CRISPR systems from Lachnospiraceae bacterium was coupled with a hybridization chain reaction (HCR) to develop an electrochemical biosensor for detecting the pathogenic bacterium, Salmonella typhimurium. Autonomous cross-opening of functional DNA hairpin structures of HCR yielded polymer double-stranded DNA wires consisting of numerous single-stranded DNAs, which initiated the trans-cleavage activity of CRISPR-Cas12a to indiscriminately cleave random single-stranded DNA labeling electrochemical tags on the surface of the electrode. It led to a variation in the electron transfer of electrochemical tags. The polymer double-stranded DNA of HCR was immobilized on dynabeads (DBs) via the S. typhimurium aptamer and released from DBs. The established method could selectively and sensitively quantify S. typhimurium in samples with detection limits of 20 CFU/mL. Our study provides a novel insight for exploring universal analytical methods for pathogenic bacteria based on CRISPR-Cas12a coupled with HCR.

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