食品中沙门氏菌DNA环介导等温扩增快速检测方法的建立  

Development of a loop-mediated isothermal amplification method with fimY gene for rapid detection of Salmonella in food

作　　者：徐义刚 崔丽春[2] 杨君宏 李苏龙 李丹丹[4] 姜艳春 张子群 

机构地区：[1]黑龙江出入境检验检疫局检验检疫技术中心,黑龙江哈尔滨150001 [2]东北林业大学,黑龙江哈尔滨150040 [3]哈尔滨市粮食研究所,黑龙江哈尔滨150076 [4]海南出入境检验检疫局检验检疫技术中心,海南海口570125

出　　处：《中国兽医科学》2010年第5期452-458,共7页Chinese Veterinary Science

基　　金：国家质检总局科技计划项目(2008IK162);黑龙江省博士后基金项目(LBH-Z09)

摘　　要：根据沙门氏菌属高度保守的fimY基因序列设计了2对特异性引物,采用环介导等温扩增方法(LAMP)建立了食品中沙门氏菌的LAMP快速检测方法。结果表明,建立的LAMP方法具有高度特异性,经对40种共68株细菌进行扩增,所试32株沙门氏菌均为LAMP阳性,其他菌株为LAMP阴性。建立的LAMP方法对培养的沙门氏菌的检测灵敏度为5CFU/管,对污染食品中沙门氏菌的检测灵敏度为9CFU/管,60min内即可完成检测。用建立的LAMP方法对802份肉类、蛋、奶及其制品和动物腹泻物、人工污染样品等进行检测,共检出165份LAMP阳性样本,与国标(GB/T4789.4-2008)方法的检测结果一致。建立的LAMP方法操作简便、特异性强、灵敏度高,具有良好的实用性。According to the highly conserved fimY gene of Salmonella,two pairs of primers were designed and the method for detecting Salmonella in food was developed with loop-mediated isothermal amplification（LAMP）.In results,sixty-eight strains of forty bacteria species were used for specificity test and all the tested Salmonella strains were positive and the other strains were negative in LAMP,indicating that the LAMP method was highly specific for target bacteria with the sensitivity of 5 CFU per test tube for cultivated Salmonella and 9 CFU per test tube for its contaminated food.The LAMP test could be completed in 60min.165 from 802 samples of meat,egg,milk and its products,animal diarrhea materials and artificial contamination samples were positive in LAMP,which was in accordance with the testing result according to GB/T4789.4-2008.The results showed that the LAMP assay was a sensitive,rapid and simple tool for detecting Salmonella.

关 键 词：沙门氏菌 fimY基因 环介导等温扩增 快速检测 

分 类 号：S859.84[农业科学—临床兽医学]