沙门氏菌
环介导等温扩增
肠沙门氏菌
检出限
生物
聚合酶链反应
血清型
污染
基因
DNA
微生物学
计算生物学
分子生物学
化学
色谱法
细菌
遗传学
生态学
作者
Yuting Shang,Qinghua Ye,Shuzhen Cai,Qingping Wu,Rui Pang,Shuanghong Yang,Xinran Xiang,Chu‐Fang Wang,Fei Zha,Yu Ding,Yinzhi Zhang,Jiasheng Wang,Xiulan Sun,Jumei Zhang
标识
DOI:10.1016/j.lwt.2021.110999
摘要
Various molecular techniques have been introduced for Salmonella detection. Loop-mediated isothermal amplification (LAMP) is a simple and easy-to-operate detection method compared with conventional PCR. However, the specificity and efficiency of DNA amplification relies on the selected target sequence. In this study, we aimed to identify new molecular targets and develop a LAMP-based method for the rapid detection of Salmonella in food samples. Using bioinformatics analysis and PCR verification, 3 and 10 molecular targets respectively unique to Salmonella genus and S. enterica were identified. The limit of detection (LOD) for these targets ranged from 2.1 × 102 to 2.1 × 103 CFU/mL. Of these, the ssaQ gene yielded 100% positive results upon testing with 23 most commonly reported Salmonella serovars and showed no cross-reaction with 22 non-Salmonella strains. Furthermore, its LOD was 2.1 × 101 CFU/mL, making it 10-fold more sensitive than the optimal PCR-based method. Therefore, the ssaQ gene was selected as the target gene to detect Salmonella using LAMP. The ability of the ssaQ-based LAMP method of detecting Salmonella in food samples was consistent with that of the standard culture method. The developed method has potential applications in identifying Salmonella contamination in food as well as for rapid and precise tracing of contamination sources in clinical diagnostics.
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