Xiao Yiran, Ren Honglin, Hu Pan, Wang Yang, Wang Han, Li Yansong, Feng Kai, Wang Cong, Cao Qi, Guo Yuxi, Liu Zengshan, Lu Shiying
State Key Laboratory for Zoonotic Diseases, Key Laboratory for Zoonosis Research of the Ministry of Education, Institute of Zoonosis, College of Veterinary Medicine, Jilin University, Changchun 130062, China.
Jilin Province Positioning Slaughter Management Office, Xi'an Road, Changchun 130062, China.
Foods. 2022 Jul 21;11(14):2160. doi: 10.3390/foods11142160.
is a dangerous foodborne human pathogen that mainly causes gastroenteritis. Ideal methods for the detection of pathogens in food should be rapid, sensitive, specific, and cost effective. To this end, novel in vitro nucleic acid identification methods based on clustered, regularly interspaced short palindromic repeats (CRISPR)-associated protein (Cas) endonuclease have received increasing attention. In this study, a simple, visual, and ultrasensitive method, based on CRISPR/Cas12a with recombinase polymerase amplification (RPA), was developed for the detection of . The results show that a specific attachment invasion locus gene () can be rapidly detected using a CRISPR/Cas12a-RPA-based system. Application of the method to raw pork, which was artificially infected with , achieved an estimated detection limit of 1.7 CFU/mL in less than 45 min, and this was 100 times lower compared with qPCR. The results indicated that the CRISPR/Cas12a-RPA system has good potential for monitoring pathogenic in the chilled meat supply chain.
是一种危险的食源性人类病原体，主要引起肠胃炎。食品中病原体检测的理想方法应快速、灵敏、特异且具有成本效益。为此，基于成簇规律间隔短回文重复序列（CRISPR）相关蛋白（Cas）核酸内切酶的新型体外核酸鉴定方法受到越来越多的关注。在本研究中，开发了一种基于CRISPR/Cas12a与重组酶聚合酶扩增（RPA）的简单、可视化且超灵敏的方法用于检测。结果表明，使用基于CRISPR/Cas12a-RPA的系统可以快速检测特定的附着侵袭位点基因（）。将该方法应用于人工感染的生猪肉，在不到45分钟内实现了估计1.7 CFU/mL的检测限，这比qPCR低100倍。结果表明，CRISPR/Cas12a-RPA系统在冷藏肉供应链中监测致病性方面具有良好的潜力。
