A one-pot method based on rolling circle amplification and light-activated CRISPR/Cas12a reaction for simple and highly sensitive detection of Staphylococcus aureus

作者全名："Wu, Qian; Jiang, Song; Huang, Yong; Zhang, Lijun; Li, Zongying; Hou, Yongbin; Zhang, Juan; Wang, Yuwei; Zhu, Chuanlin; Zhou, Dandan; Chen, Weixian; Li, Dandan"

作者地址："[Wu, Qian; Jiang, Song; Huang, Yong; Zhang, Lijun; Li, Zongying; Hou, Yongbin; Chen, Weixian; Li, Dandan] Chongqing Med Univ, Affiliated Hosp 2, Dept Lab Med, Chongqing 400016, Peoples R China; [Zhang, Juan; Wang, Yuwei; Zhu, Chuanlin; Zhou, Dandan] Chongqing Hosp Tradit Chinese Med, Lab Diag & Treatment Infect Dis Integrated Tradit, Dept Lab Med, Chongqing 400016, Peoples R China"

通信作者："Chen, WX; Li, DD (通讯作者)，Chongqing Med Univ, Affiliated Hosp 2, Dept Lab Med, Chongqing 400016, Peoples R China."

来源：CHEMICAL ENGINEERING JOURNAL

ESI学科分类：ENGINEERING

WOS号：WOS:001105693800001

JCR分区：Q1

影响因子：13.3

年份：2023

卷号：477

期号：　

开始页：　

结束页：　

文献类型：Article

关键词：One-pot method; Light-activated CRISPR/Cas12a; Rolling circle amplification; Staphylococcus aureus

摘要："Developing robust methods that achieve simple and highly sensitive detection of Staphylococcus aureus (S. aureus) is still a challenge. Isothermal nucleic acid amplification reactions-triggered CRISPR/Cas12a strategies offer promising sensing tools for bacteria detection, but have several issues that need to be solved, such as incompatible reaction systems, compromised sensitivity, and multiple reaction steps. Here, we develop a light activated one-pot method based on rolling circle amplification-triggered CRISPR/Cas12a (RCA-Cas12a) for analyzing S. aureus directly. In the one-pot assay, the Cas12a reaction is started under light irradiation only after the RCA reaction is completed, thus avoiding interference between the two systems. The light-activated method is simple to operate, and its sensitivity is two orders of magnitude higher than that of the traditional one-pot assay. Accurately identifying contaminated samples from 120 real samples (serum, milk, and broth) prepared in a double-blind way demonstrates the good application capacity of the light-activated one-pot method. The method has a shorter analysis time, fewer operation steps, and higher accuracy in identifying contaminated samples compared with bacterial culture. Thus, the light-activated one-pot RCA-Cas12a method represents a simple tool for sensitive and specific detection of S. aureus in food and clinical samples."

基金机构：Natural Science Foundation Project of Chongqing [cstc2020jcyj-msxmX0190]; Chongqing Medical Scientific Research Project-Joint project of Chongqing Health Commission; Science and Technology Bureau [2023MSXM035]; Chongqing scientific research institutions performance incentive and guidance project [cstc2021jxjl130038]; Kuanren Talents Program of the second affiliated hospital of Chongqing Medical University [kryc-yq-2117]

基金资助正文："This work was financially supported by the Natural Science Foundation Project of Chongqing (cstc2020jcyj-msxmX0190) , Chongqing Medical Scientific Research Project-Joint project of Chongqing Health Commission and Science and Technology Bureau (2023MSXM035) , Chongqing scientific research institutions performance incentive and guidance project (cstc2021jxjl130038) , and Kuanren Talents Program of the second affiliated hospital of Chongqing Medical University (kryc-yq-2117)."