Next Article in Journal
Decoding Flavonoid Metabolism for Nutritional Enhancement: A Transcriptome–Metabolome Integration Study of Biosynthesis in Edible Chrysanthemum indicum L.
Previous Article in Journal
Flavor Quality and Lipid-Lowering Function of Mixed Fermented Pu-erh Tea with Various Monascus Species
 
 
Font Type:
Arial Georgia Verdana
Font Size:
Aa Aa Aa
Line Spacing:
Column Width:
Background:
This is an early access version, the complete PDF, HTML, and XML versions will be available soon.
Article

CRISPR-Cas12a/Aurora Deoxyribozyme Cascade: A Label-Free Ultrasensitive Platform for Rapid Salmonella Detection

1
Key Laboratory of Environmental Pollution Monitoring and Disease Control, Ministry of Education, School of Public Health, Guizhou Medical University, No. 6 Ankang Road, Guian New Area, Guizhou 561113, China
2
Institute of Quality Standard and Testing Technology, Beijing Academy of Agriculture and Forestry Sciences, Beijing 100097, China
*
Authors to whom correspondence should be addressed.
These authors contributed equally to this work.
Foods 2025, 14(11), 1892; https://doi.org/10.3390/foods14111892
Submission received: 15 April 2025 / Revised: 18 May 2025 / Accepted: 24 May 2025 / Published: 26 May 2025
(This article belongs to the Section Food Nutrition)

Abstract

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 × 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.
Keywords: nucleic acid detection; fluorescence sensing; food safety; DNAzyme; pathogen identification nucleic acid detection; fluorescence sensing; food safety; DNAzyme; pathogen identification

Share and Cite

MDPI and ACS Style

Shi, C.; Tan, H.; Yu, Z.; Li, W.; Man, Y.; Zhang, Q. CRISPR-Cas12a/Aurora Deoxyribozyme Cascade: A Label-Free Ultrasensitive Platform for Rapid Salmonella Detection. Foods 2025, 14, 1892. https://doi.org/10.3390/foods14111892

AMA Style

Shi C, Tan H, Yu Z, Li W, Man Y, Zhang Q. CRISPR-Cas12a/Aurora Deoxyribozyme Cascade: A Label-Free Ultrasensitive Platform for Rapid Salmonella Detection. Foods. 2025; 14(11):1892. https://doi.org/10.3390/foods14111892

Chicago/Turabian Style

Shi, Cong, Huimin Tan, Zhou Yu, Weilin Li, Yan Man, and Qinghai Zhang. 2025. "CRISPR-Cas12a/Aurora Deoxyribozyme Cascade: A Label-Free Ultrasensitive Platform for Rapid Salmonella Detection" Foods 14, no. 11: 1892. https://doi.org/10.3390/foods14111892

APA Style

Shi, C., Tan, H., Yu, Z., Li, W., Man, Y., & Zhang, Q. (2025). CRISPR-Cas12a/Aurora Deoxyribozyme Cascade: A Label-Free Ultrasensitive Platform for Rapid Salmonella Detection. Foods, 14(11), 1892. https://doi.org/10.3390/foods14111892

Note that from the first issue of 2016, this journal uses article numbers instead of page numbers. See further details here.

Article Metrics

Back to TopTop