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A Dual-mode platform for the rapid detection of Escherichia coli O157:H7 based on CRISPR/Cas12a and RPA

文献类型： 外文期刊

作者： Luo, Jiawei 1 ; Xu, Danhong 2 ; Wang, Jinbin 2 ; Liu, Hua 2 ; Li, You 2 ; Zhang, Yan 1 ; Zeng, Haijuan 2 ; Deng, Bo 6 ; Liu, Xiaofeng 1 ;

作者机构： 1.Lanzhou Univ Technol, Sch Life Sci & Engn, Lanzhou, Peoples R China

2.Shanghai Acad Agr Sci, Biotechnol Res Inst, Key Lab Agr Genet & Breeding, Shanghai, Peoples R China

3.Minist Agr & Rural Affairs, Crops Ecol Environm Secur Inspect & Supervis Ctr S, Shanghai, Peoples R China

4.Shanghai Coelite Agr Scitech Grp Co Ltd, Shanghai, Peoples R China

5.Changsha Med Univ, Sch Publ Hlth, Academician Workstat, Changsha, Peoples R China

6.Shanghai Ctr Agri Prod Qual & Safety, Shanghai, Peoples R China

关键词： Escherichia coli O157:H7; Recombinant polymerase amplification; CRISPR/Cas12a; Lateral flow chromatography; Fluorescence detection

期刊名称：ANALYTICAL AND BIOANALYTICAL CHEMISTRY （ 影响因子：4.3； 五年影响因子：4.0 ）

ISSN： 1618-2642

年卷期： 2024 年

页码：

收录情况： SCI

摘要： Escherichia coli O157:H7 (E. coli O157:H7) is a foodborne pathogenic microorganism that is commonly found in the environment and poses a significant threat to human health, public safety, and economic stability worldwide. Thus, early detection is essential for E. coli O157:H7 control. In recent years, a series of E. coli O157:H7 detection methods have been developed, but the sensitivity and portability of the methods still need improvement. Therefore, in this study, a rapid and efficient testing platform based on the CRISPR/Cas12a cleavage reaction was constructed. Through the integration of recombinant polymerase amplification and lateral flow chromatography, we established a dual-interpretation-mode detection platform based on CRISPR/Cas12a-derived fluorescence and lateral flow chromatography for the detection of E. coli O157:H7. For the fluorescence detection method, the limits of detection (LODs) of genomic DNA and E. coli O157:H7 were 1.8 fg/mu L and 2.4 CFU/mL, respectively, within 40 min. Conversely, for the lateral flow detection method, LODs of 1.8 fg/mu L and 2.4 x 10(2) CFU/mL were achieved for genomic DNA and E. coli O157:H7, respectively, within 45 min. This detection strategy offered higher sensitivity and lower equipment requirements than industry standards. In conclusion, the established platform showed excellent specificity and strong universality. Modifying the target gene and its primers can broaden the platform's applicability to detect various other foodborne pathogens.

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