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食品中沙门氏菌LAMP快速检测方法的建立
引用本文:黄金海,孙跃辉,陈瑞,庄世文,刘莹,王静思. 食品中沙门氏菌LAMP快速检测方法的建立[J]. 天津大学学报(自然科学与工程技术版), 2012, 0(5): 468-472
作者姓名:黄金海  孙跃辉  陈瑞  庄世文  刘莹  王静思
作者单位:天津大学化工学院;华南农业大学动物医学院
基金项目:天津市应用基础及前沿技术研究计划项目(08JCZDJC22600)
摘    要: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.

关 键 词:沙门氏菌  侵袭性因子A  环介导等温扩增技术

Development of Loop-Mediate Isothermal Amplification Assay for Salmonella Detection in Food
HUANG Jin-hai,SUN Yue-hui,CHEN Rui,ZHUANG Shi-wen,LIU Ying,WANG Jing-si. Development of Loop-Mediate Isothermal Amplification Assay for Salmonella Detection in Food[J]. Journal of Tianjin University(Science and Technology), 2012, 0(5): 468-472
Authors:HUANG Jin-hai  SUN Yue-hui  CHEN Rui  ZHUANG Shi-wen  LIU Ying  WANG Jing-si
Affiliation:1(1.School of Chemical Engineering and Technology,Tianjin University,Tianjin 300072,China;2.College of Veterinary Medicine,South China Agricultural University,Guangzhou 510642,China)
Abstract: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.
Keywords:Salmonella  invA  loop-mediated isothermal amplification
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