CRISPR-Cas12a/Aurora Deoxyribozyme Cascade: A Label-Free Ultrasensitive Platform for Rapid Salmonella Detection
文献类型： 外文期刊
作者： Shi, Cong 1 ; Tan, Huimin 1 ; Yu, Zhou 1 ; Li, Weilin 1 ; Man, Yan 1 ; Zhang, Qinghai 1 ;
作者机构： 1.Guizhou Med Univ, Sch Publ Hlth, Minist Educ, Key Lab Environm Pollut Monitoring & Dis Control, 6 Ankang Rd Guian New Area, Guiyang 561113, Guizhou, Peoples R China
2.Beijing Acad Agr & Forestry Sci, Inst Qual Stand & Testing Technol, Beijing 100097, Peoples R China
关键词： nucleic acid detection; fluorescence sensing; food safety; DNAzyme; pathogen identification
期刊名称：FOODS （ 影响因子：5.1； 五年影响因子：5.6 ）
ISSN：
年卷期： 2025 年 14 卷 11 期
页码：
收录情况： SCI
摘要： The rapid and ultrasensitive detection of Salmonella holds strategic significance for food safety surveillance and public health protection systems. This study innovatively developed a label-free biosensing platform based on the synergistic integration of Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)-Cas12a and the fluorescent deoxyribozyme Aurora for the efficient detection of foodborne Salmonella. The detection mechanism operates through a molecular cascade reaction: target-activated Cas12a protein specifically degrades Aurora deoxyribozyme via its trans-cleavage activity, thereby abolishing the enzyme's catalytic capability to convert 4-methylumbelliferyl phosphate (4-MUP) into the highly fluorescent product 4-methylumbelliferone (4-MU). This cascade ultimately enables quantitative target analysis through fluorescence signal attenuation. Following systematic optimization of critical reaction parameters, the biosensing system demonstrated exceptional analytical performance: a detection limit of 1.29 CFU/mL with excellent linearity (R2 = 0.992) spanning six orders of magnitude (1.65 x 101-106 CFU/mL), along with high specificity against multiple interfering bacterial strains. Spike-and-recovery tests in complex food matrices (milk, chicken, and lettuce) yielded recoveries of 90.91-99.40% (RSD = 3.55-4.72%), confirming robust practical applicability. Notably, the platform design allows flexible detection of other pathogens through simple replacement of CRISPR guide sequences.
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