期刊
LETTERS IN APPLIED MICROBIOLOGY
卷 75, 期 4, 页码 899-907出版社
OXFORD UNIV PRESS
DOI: 10.1111/lam.13765
关键词
CRISPR; Cas12a; detection; optimized crRNA; RPA; Salmonella
资金
- Young Talent Development Plan of Changzhou Health Commission [CZQM2020113]
- Science and Technology Application Basic Plan of Changzhou (Medium Subsidized) [CJ20200105, CJ20210100, CJ20210073]
- Jiangsu Provincial Medical Youth Talent [QNRC2016309]
A simple, rapid, sensitive, and highly accurate detection technique for Salmonella based on CRISPR-Cas12a was developed, which can rapidly detect trace Salmonella through fluorescence intensity.
Salmonella is a crucial food-borne pathogen causing food poisoning, leading to severe public health events. Here, we developed a technique by integrating recombinase polymerase amplification with CRISPR-LbCas12a and employing two targets with engineered crRNA for detection of Salmonella (RPA-LbCas12a-TTECDS). Our findings revealed that this novel method rapidly detects trace Salmonella in food through fluorescence intensity and provides a template for other food-borne pathogen detection methods. Further, crRNA was optimized to increase detection sensitivity. Double targets were used to enhance the detection accuracy, reaching the level of qPCR, which was superior to fluorescent RPA. The RPA-LbCas12a-TTECDS system specifically detected Salmonella levels as low as 50 CFU per ml at 37 degrees C in 1 h. In summary, a simple, rapid, sensitive and high accuracy detection technique based on CRISPR-Cas12a was created for Salmonella detection without complicated equipment.
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