Generation and application of a novel high-throughput detection based on RPA-CRISPR technique to sensitively monitor pathogenic microorganisms in the environment
文献类型：期刊论文
| 作者 | Liu, Li1,2; Duan, Jin-Jing3; Wei, Xing-Yi1,4; Hu, Huan1,4; Wang, Yuan-Bo1,4; Jia, Pan-Pan3; Pei, De-Sheng3
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| 刊名 | SCIENCE OF THE TOTAL ENVIRONMENT
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| 出版日期 | 2022-09-10 |
| 卷号 | 838页码:9 |
| ISSN号 | 0048-9697 |
| 关键词 | Nucleic acid detection CRISPR Cas12a Staphylococcus aureus RPA-CRISPR Cas12a-FQ Cas12a-LFA |
| DOI | 10.1016/j.scitotenv.2022.156048 |
| 通讯作者 | Pei, De-Sheng(peids@cqmu.edu.cn) |
| 英文摘要 | Staphylococcus aureus (S. aureus) is an important opportunistic human and animal pathogen that can cause a wide diversity of infections. Due to its environmental health risks, it is crucial to establish a time-saving, high-throughput, and highly sensitive technique for water quality surveillance. In this study, we developed a novel method to detect S. aureus in the water environment based on recombinase polymerase amplification (RPA) and CRISPR/Cas12a. This method utilizes isothermal amplification of nucleic acids and the trans-cleavage activity of the CRISPR/Cas12a system to generate fluorescence signals with a single-stranded DNA-fluorophore-quencher (ssDNA-FQ) reporter and a naked-eye detected lateral flow assay (LFA). Our RPA-CRISPR/Cas12a detection system can reduce the detection time to 35 min and enhance the high-throughput detection threshold to >= 5 copies of pathogen DNA, which is more sensitive than that of reported. Moreover, in the lower reaches of the Jialing River in Chongqing, China, 10 water samples from the mainstream and 7 ones from tributaries were successfully monitored S. aureus for less than 35 min using RPA-CRISPR/Cas12a detection system. Taken together, a novel high-throughput RPA-CRISPR detection was established and firstly applied for sensitively monitoring S. aureus in the natural water environment. |
| 资助项目 | High-level Talents Project of Chong-qing Medical University[R4014] ; CAS Team Project of the Belt and Road ; Research Program of Chongqing Science and Technology Commission[cstc2019jcyj-zdxmX0035] ; Research Program of Chongqing Science and Technology Commission[CSTCCXLJRC201714] ; Program of China-Sri Lanka Joint Center for Water Technology Research and Demonstration by Chinese Academy of Sciences (CAS) /China-Sri Lanka Joint Center for Education and Research by CAS ; International Partnership Program of CAS[121311kysb20190071] |
| WOS研究方向 | Environmental Sciences & Ecology |
| 语种 | 英语 |
| 出版者 | ELSEVIER |
| WOS记录号 | WOS:000806111000005 |
| 源URL | [http://119.78.100.138/handle/2HOD01W0/16305]  |
| 专题 | 中国科学院重庆绿色智能技术研究院 |
| 通讯作者 | Pei, De-Sheng |
| 作者单位 | 1.Chinese Acad Sci, Univ Chinese Acad ofSciences, Chongqing Sch, Chongqing Inst Green & Intelligent Technol, Chongqing 400714, Peoples R China 2.Univ Chinese Acad Sci, Beijing 100049, Peoples R China 3.Chongqing Med Univ, Sch Publ Hlth & Management, Chongqing 400016, Peoples R China 4.Chongqing Jiaotong Univ, Chongqing 400074, Peoples R China |
| 推荐引用方式 GB/T 7714 | Liu, Li,Duan, Jin-Jing,Wei, Xing-Yi,et al. Generation and application of a novel high-throughput detection based on RPA-CRISPR technique to sensitively monitor pathogenic microorganisms in the environment[J]. SCIENCE OF THE TOTAL ENVIRONMENT,2022,838:9. |
| APA | Liu, Li.,Duan, Jin-Jing.,Wei, Xing-Yi.,Hu, Huan.,Wang, Yuan-Bo.,...&Pei, De-Sheng.(2022).Generation and application of a novel high-throughput detection based on RPA-CRISPR technique to sensitively monitor pathogenic microorganisms in the environment.SCIENCE OF THE TOTAL ENVIRONMENT,838,9. |
| MLA | Liu, Li,et al."Generation and application of a novel high-throughput detection based on RPA-CRISPR technique to sensitively monitor pathogenic microorganisms in the environment".SCIENCE OF THE TOTAL ENVIRONMENT 838(2022):9. |
入库方式： OAI收割
来源：重庆绿色智能技术研究院
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