FOOD SCIENCE ›› 2025, Vol. 46 ›› Issue (23): 13-20.doi: 10.7506/spkx1002-6630-20250523-152

• Rapid Food Safety Testing • Previous Articles    

Development of a CRISPR/Cas12a-Based Fluorescence Detection Method for Pseudomonas fluorescens in Dairy Products

XIE Xinna, QIU Manyan, ZHANG Xiru, ZHU Danqing, ZHANG Jingwen, YANG Xinyan, JIANG Yujun, MAN Chaoxin, ZHANG Xianlong   

  1. (1. Key Laboratory of Dairy Science, Ministry of Education, College of Food Science, Northeast Agricultural University, Harbin 150030, China; 2. Key Laboratory of Infant Formula Food, State Administration for Market Regulation, Harbin 150030, China; 3. Food Laboratory of Zhongyuan, Luohe 462300, China)
  • Published:2025-12-26

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Abstract: In this study, by designing specific polymerase chain reaction (PCR) primers and clustered regularly interspaced short palindromic repeats (CRISPR) RNA (crRNA) probes targeting the aprX gene of Pseudomonas fluorescens, a CRISPR/CRISPR-associated protein 12a (Cas12a)-based fluorescence assay was successfully established for the rapid and sensitive detection of P. fluorescens in dairy products. The results showed that the assay exhibited a linear range of 102–108 CFU/mL, with a limit of detection (LOD) of 101 CFU/mL. Moreover, the method demonstrated excellent specificity and no cross-reactivity with common pathogens (such as Salmonella). Furthermore, recoveries from pasteurized milk and ultra-high temperature-treated milk spiked with different concentrations of P. fluorescens ranged from 102.218% to 111.981%. Overall, the CRISPR/Cas12a fluorescence assay exhibited high sensitivity and specificity, providing a novel approach for the rapid detection of P. fluorescens in dairy products.

Key words: Pseudomonas fluorescens; clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 12a; polymerase chain reaction; dairy products; rapid detection

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