沙门氏菌
清脆的
反式激活crRNA
检出限
基因组DNA
重组酶聚合酶扩增
聚合酶链反应
细菌
色谱法
化学
生物
计算生物学
重组DNA
DNA
基因
Cas9
遗传学
作者
Li Liu,Gang Zhao,Xiangmei Li,Zhen-Lin Xu,Hongtao Lei,Xing Shen
标识
DOI:10.1016/j.lwt.2022.113443
摘要
Salmonella species are common foodborne pathogenic bacteria. At present, most detection methods for Salmonella are unsuitable for on-site applications because they require large instruments or complicated procedures. This study developed a novel method of the on-site detection for Salmonella in food by combining the CRISPR/Cas12a system with recombinant polymerase amplification (RPA). The optimal concentration ratio of Cas12a enzyme, CRISPR RNA (crRNA) and FQ-probe is 1 : 1: 2.5. The detection limit of the RPA–CRISPR/Cas12a method was 1 × 10−4 ng/μL for genomic DNA (gDNA), and 102 CFU/mL for bacterial liquid. The method exhibited no cross-reactivity to 4 common pathogenic bacteria. A simple boiling method was used to pretreat chicken and egg samples to meet on-site testing requirements. The detection limits of chicken and egg samples were 103 CFU/mL initially and could reach 102 CFU/mL and 101 CFU/mL, respectively, after 3 h enrichment at 37 °C. The sensitivity of the RPA–CRISPR/Cas12a method was comparable to that of the quantitative polymerase chain reaction (qPCR) technique. In addition, the operation of this method is simpler and faster than qPCR. Thus, the method is applicable to the on-site detection for Salmonella in food.
科研通智能强力驱动
Strongly Powered by AbleSci AI