金黄色葡萄球菌
清脆的
耐甲氧西林金黄色葡萄球菌
管(容器)
微生物学
生物
细菌
材料科学
遗传学
基因
复合材料
作者
Yanan Li,Zhonglin Shi,Anzhong Hu,Jun-sheng Cui,Yang Ke,Yong Liu,Guoqing Deng,Cancan Zhu,Ling Zhu
出处
期刊:Diagnostics
[MDPI AG]
日期:2022-03-28
卷期号:12 (4): 829-829
被引量:29
标识
DOI:10.3390/diagnostics12040829
摘要
Methicillin-resistant Staphylococcus aureus (MRSA) is a severe health threat causing high-level 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.
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