One-tube RPA-CRISPR Cas12a/Cas13a rapid detection of methicillin-resistant Staphylococcus aureus
文献类型： 外文期刊
作者： Liu, Yujie 1 ; Liu, Hui 2 ; Yu, Guanliu 1 ; Sun, Wenbo 3 ; Aizaz, Muhammad 1 ; Yang, Guiwen 1 ; Chen, Lei 1 ;
作者机构： 1.Shandong Normal Univ, Coll Life Sci, Shandong Prov Key Lab Anim Resistance Biol, Jinan, Shandong, Peoples R China
2.Jinan Ctr Dis Control & Prevent, Jinan, Shandong, Peoples R China
3.Shandong Acad Agr Sci, Inst Anim Sci & Vet Med, Shandong Key Lab Anim Dis Control & Breeding, Jinan, Shandong, Peoples R China
关键词： MRSA; Staphylococcus aureus; CRISPR Cas12a/Cas13a; RPA; Nucleic acid detection
期刊名称：ANALYTICA CHIMICA ACTA （ 影响因子：6.2； 五年影响因子：5.9 ）
ISSN： 0003-2670
年卷期： 2023 年 1278 卷
页码：
收录情况： SCI
摘要： At present, methicillin-resistant Staphylococcus aureus (MRSA) has caused a serious impact on a global scale. The infection and carrier rate of MRSA in the community is increasing year by year, but there is still no convenient detection system for on-site rapid detection. It is very important to select a rapid detection system to accurately and quickly detect patients infected with MRSA. We have developed a high-efficient single-tube detection platform based on RPA and CRISPR reaction system to detect the genes of mecA and clfA of MRSA. Using this detection platform, visual MRSA detection could be achieved in 30 min. It was observed that this detection platform was capable to successfully detect the target genomic as low as 5 copies & mu;L-1, and the reaction was completed in one step without opening the lid. This detection platform could only detect MRSA, but not other common clinical pathogenic bacteria, such as Salmonella, Pseudomonas aeruginosa, Staphylococcus xylosus, Aeromonas hydrophila, Escherichia coli and Staphylococcus warneri, indicated its satisfactory selectivity for MRSA without interference from other bacteria. The results of clinical samples show that the platform has outstanding advantages in sensitivity, specificity and identification of methicillin resistance. The entire reaction can be completed in one step in the handheld instrument without opening the cover, avoiding aerosol pollution during the reaction. The detection platform combined with handheld instruments will have great application potential in point-of-care testing.
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