Isothermal Amplification and CRISPR/Cas12a-System-Based Assay for Rapid, Sensitive and Visual Detection of Staphylococcus aureus
文献类型： 外文期刊
作者： Xu, Danhong 1 ; Zeng, Haijuan 2 ; Wu, Wenhui 1 ; Liu, Hua 2 ; Wang, Jinbin 2 ;
作者机构： 1.Shanghai Ocean Univ, Sch Food Sci & Technol, Shanghai 201499, Peoples R China
2.Shanghai Acad Agr Sci, Biotechnol Res Inst, Key Lab Agr Genet & Breeding, Shanghai 201106, Peoples R China
3.Minist Agr & Rural Affairs, Crops Ecol Environm Secur Inspect & Supervis Ctr, Shanghai 201106, Peoples R China
关键词： S. aureus; LAMP; RPA; immunochromatographic test strip; fluorescence detection
期刊名称：FOODS （ 影响因子：5.2； 五年影响因子：5.5 ）
ISSN：
年卷期： 2023 年 12 卷 24 期
页码：
收录情况： SCI
摘要： Staphylococcus aureus exists widely in the natural environment and is one of the main food-borne pathogenic microorganisms causing human bacteremia. For safe food management, a rapid, high-specificity, sensitive method for the detection of S. aureus should be developed. In this study, a platform for detecting S. aureus (nuc gene) based on isothermal amplification (loop-mediated isothermal amplification-LAMP, recombinase polymerase amplification-RPA) and the clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR-associated (Cas12a) proteins system (LAMP, RPA-CRISPR/Cas12a) was proposed. In this study, the LAMP, RPA-CRISPR/Cas12a detection platform and immunochromatographic test strip (ICS) were combined to achieve a low-cost, simple and visualized detection of S. aureus. The limit of visual detection was 57.8 fg/mu L of nuc DNA and 6.7 x 10(2) CFU/mL of bacteria. Moreover, the platform could be combined with fluorescence detection, namely LAMP, RPA-CRISPR/Cas12a-flu, to establish a rapid and highly sensitive method for the detection of S. aureus. The limit of fluorescence detection was 5.78 fg/mu L of genomic DNA and 67 CFU/mL of S. aureus. In addition, this detection platform can detect S. aureus in dairy products, and the detection time was similar to 40 min. Consequently, the isothermal amplification CRISPR/Cas12a platform is a useful tool for the rapid and sensitive detection of S. aureus in food.
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