食品中沙门氏菌LAMP快速检测方法的建立

Development of Loop-Mediate Isothermal Amplification Assay for Salmonella Detection in Food

食品及其原料中沙门氏菌的快速、现场检测对食品安全控制具有重要意义.根据沙门氏菌invA基因核苷酸序列设计一组引物,应用环介导等温扩增技术(LAMP),分别对7种不同血清型沙门氏菌和4种非沙门氏菌进行扩增,同时建立沙门氏菌人工污染的食品模型,比较了LAMP法与活菌计数检测的敏感性.结果表明,该方法仅对沙门氏菌产生特异性扩增,灵敏度高达336,mL-1,食品样品经细菌富集培养后,检测灵敏度高达8.25,g-1.所建立的检测沙门氏菌方法具有较高的特异性与灵敏性,操作简单、快速,可用于沙门氏菌污染食品的快速检测.

It is important to detect Salmonella in food or food materials in order to assure the food safety. A rapid sen- sitive loop-mediated isothermal amplification （LAMP） method assay for the food-borne pathogen Salmonella detection was developed. A set of primers were designed according to the nucleotide sequence of the target invA gene in Salmo- nella, 7 strains of Salmonella and 4 non-Salmonella bacteria were detected by LAMP, and meanwhile the mode of artificially contaminated food was constructed to evaluate the sensitivity of LAMP assay and bacterial cell counting method, The results showed that all the 7 Salmonella bacterial strains had specific amplification, but the 4 non- Salmonella bacterial strains submitted negative reactions. Sensitivity of LAMP assay for pure cell culture of Salmo- nella was 336 mL^-1. However, an 8.25 g^-1 detection limit was obtained for Salmonella artificially contaminated food following a bacterial culture enrichment process, while there was no amplification from the negative control. In con- clusion, the LAMP assay developed in the present study is a specific, sensitive, simple and convenient method for the rapid detection of Salmonella in contaminated foods.