Rapid One-Tube RPA-CRISPR/Cas12 Detection Platform for Methicillin-Resistant Staphylococcus aureus

doi:10.3390/diagnostics12040829

	Rapid One-Tube RPA-CRISPR/Cas12 Detection Platform for Methicillin-Resistant Staphylococcus aureus
	Li, Yanan 1,2; Shi, Zhonglin 1,3; Hu, Anzhong 1; Cui, Junsheng 1; Yang, Ke 1; Liu, Yong 1; Deng, Guoqing 1; Zhu, Cancan 1; Zhu, Ling 1
刊名	DIAGNOSTICS
	2022-04-01
卷号	12
关键词	methicillin-resistant Staphylococcus aureus mecA gene RPA-CRISPR/Cas12a one-tube POCT
DOI	10.3390/diagnostics12040829
通讯作者	Zhu, Cancan(zhucancan@aiofm.ac.cn) ; Zhu, Ling(zhul@aiofm.ac.cn)
英文摘要	Methicillin-resistant Staphylococcus aureus (MRSA) is a severe health threat causing highlevel morbidity and mortality in health care environments and in community settings. Though existing diagnostic methods, including PCR and culture-based methods, are routinely used in clinical practice, they are not appropriate for rapid point-of-care testing (POCT). Recently, since the development of the CRISPR/Cas technology, new possibilities for rapid point-of-care detection have emerged. In this study, we developed a rapid, accurate, and contamination-free platform for MRSA detection by integrating recombinase polymerase amplification (RPA) with the Cas12 system into one tube. Using this approach, visual MRSA detection could be achieved in 20 min. Based on the one-tube RPA-CRISPR/Cas12a platform, the assay results are visualized by lateral flow test strips (LFS) and fluorescent-based methods, including real-time and end-point fluorescence. This platform allows specific MRSA detection with a sensitivity of 10 copies for the fluorescence method and a range of 10-100 copies for the LFS. The results of 23 samples from clinical MRSA isolates showed that the coincidence rate was 100% and 95.7% of the fluorescence method and LFS, respectively, compared to qPCR. In conclusion, the one-tube RPA-CRISPR/Cas12a platform is an effective method for MRSA detection with significant potential in future practical POCT applications.
资助项目	National Natural Science Foundation of China[82002189] ; Natural Science Foundation of Anhui Province of China[2008085QF311] ; Key Research and Development Program of Anhui Province of China[202004d07020014] ; Youth Innovation Promotion Association of the Chinese Academy of Sciences[2018489] ; Youth Innovation Promotion Association of the Chinese Academy of Sciences[2021442] ; Dean's Fund of Hefei Institute of Physical Science, China[YZJJ2020QN35] ; Dean's Fund of Hefei Institute of Physical Science, China[YZJJ2021QN03]
WOS关键词	NUCLEIC-ACID DETECTION ; EPIDEMIOLOGY ; INFECTIONS
WOS研究方向	General & Internal Medicine
语种	英语
出版者	MDPI
WOS记录号	WOS:000786055700001
资助机构	National Natural Science Foundation of China ; Natural Science Foundation of Anhui Province of China ; Key Research and Development Program of Anhui Province of China ; Youth Innovation Promotion Association of the Chinese Academy of Sciences ; Dean's Fund of Hefei Institute of Physical Science, China
内容类型	期刊论文
源URL	[http://ir.hfcas.ac.cn:8080/handle/334002/128537]
专题	中国科学院合肥物质科学研究院
通讯作者	Zhu, Cancan; Zhu, Ling
作者单位	1.Chinese Acad Sci, Hefei Inst Phys Sci, Hefei 230031, Peoples R China 2.Univ Sci & Technol China, Sci Isl Branch, Grad Sch, Hefei 230026, Peoples R China 3.Anhui Univ, Inst Phys Sci & Informat Technol, Hefei 230601, Peoples R China
推荐引用方式 GB/T 7714	Li, Yanan,Shi, Zhonglin,Hu, Anzhong,et al. Rapid One-Tube RPA-CRISPR/Cas12 Detection Platform for Methicillin-Resistant Staphylococcus aureus[J]. DIAGNOSTICS,2022,12.
APA	Li, Yanan.,Shi, Zhonglin.,Hu, Anzhong.,Cui, Junsheng.,Yang, Ke.,...&Zhu, Ling.(2022).Rapid One-Tube RPA-CRISPR/Cas12 Detection Platform for Methicillin-Resistant Staphylococcus aureus.DIAGNOSTICS,12.
MLA	Li, Yanan,et al."Rapid One-Tube RPA-CRISPR/Cas12 Detection Platform for Methicillin-Resistant Staphylococcus aureus".DIAGNOSTICS 12(2022).