A novel visual loop-mediated isothermal amplification assay targeting gene62181533 for the detection of Salmonella spp. in foods
A novel visual loop-mediated isothermal amplification assay targeting gene62181533 for the detection of Salmonella spp. in foods
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DOI：
10.1016/j.foodcont.2015.07.036
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发表时间：
2016-02-01
期刊：
影响因子：
6
通讯作者：
Zhao, Haizhen
中科院分区：
文献类型：
--
作者：
Li, Junjie;Zhai, Ligong;Zhao, Haizhen
在这项研究中，我们建立了一个快速和敏感的环介导等温扩增（LAMP）检测沙门氏菌基因62181533为目标。向反应体系中加入锰离子和钙黄绿素允许在扩增反应期间观察到明显的颜色变化。使用六引物组合的LAMP的检测极限为1.057拷贝/μ l（标准质粒）、4.1CFU/ml（细胞）和7.68fg/μ l（基因组）。基于32种沙门氏菌菌株和25种非沙门氏菌细菌，LAMP测定具有100%特异性。在接种的牛奶中，在缓冲蛋白胨水中预富集12小时后，LAMP试验检测到初始接种浓度为0.81 CFU/ml的沙门氏菌。100个食品样品（牛肉，鸡肉和猪肉）进行了测试的LAMP检测。LAMP法阳性率为11/100，常规PCR法为11/100，培养法为11/100。结果表明，基于基因62181533的LAMP方法是一种快速、特异、灵敏的检测食品中沙门氏菌的方法。(c)2015爱思唯尔有限公司版权所有。
In this study, we developed a rapid and sensitive loop-mediated isothermal amplification (LAMP) assay for Salmonella detection targeting the gene62181533. The addition of manganous ions and calcein to the reaction system allowed the visualization of an apparent color change during the amplification reaction. The detection limit of LAMP using a six-primer combination was 1.057 copies/mu l (standard plasmid), 4.1 CFU/ml (cell), and 7.68 fg/mu l (genome). The LAMP assay was 100% specific based on 32 Salmonella strains and 25 non-Salmonella bacteria. In inoculated milk, the LAMP assay detected Salmonella at an initial inoculation concentration of 0.81 CFU/ml following 12 h of pre-enrichment in buffered peptone water. One hundred food samples (beef, chicken, and pork) were tested by the LAMP assay. The positive-sample ratios were 11/100 by LAMP, 11/100 by conventional PCR, and 11/100 by a culture-based method. The results revealed that the gene62181533-based LAMP assay is a rapid, specific, and sensitive method for the detection of Salmonella in foods. (c) 2015 Elsevier Ltd. All rights reserved.