沙门菌夹心DNA杂交检测方法的建立  

Establishment of a sandwich DNA hybridization assay for detection of Salmonella

作　　者：刘亚娟[1,2] 聂福平[2,3] 张超[2] 杨俊[2,3] 王昱[2,3] 石宝石 保雨 王国民[2] 李贤良[2] 李应国[2] 刘力[1] 

机构地区：[1]西南大学动物科技学院,重庆400715 [2]重庆出入境检验检疫局重庆市进出口食品安全工程中心,重庆400020 [3]重庆大学生物工程学院,重庆400030

出　　处：《中国兽医科学》2015年第12期1260-1265,共6页Chinese Veterinary Science

基　　金：质检公益性行业科研专项(201310093);国家质检总局科技项目(2014IK247,2015IK332)

摘　　要：为建立简便快速、特异检测沙门菌的方法,根据沙门菌invA基因序列,分别设计特异的捕获探针和检测探针,通过探针与目标rRNA结合,优化反应条件,建立了检测沙门菌的夹心DNA杂交方法。该方法在1.5h内即可完成全过程检测,加入TMB显色液可直接肉眼观察结果或采用酶标仪测定D450nm值判定结果。该方法对其他相关病原菌均无特异性反应,最低检测限为1.2CFU/mL,具有良好的精确度。批内变异系数在16.53%~17.97%之间,批间变异系数在16.33%~22.10%之间。本方法的建立为沙门菌的大批量、快速检测和防控提供了新的技术手段。A novel DNA hybridization technology was established for the rapid and specific detection of Salmonella using a capture probe and a detector probe,respectively,which were targeted to the invAgene.The process was completed in less than 1.5hours and the result was determined by visual inspection after adding soluble TMB substrate solution or was tested the D450 nmvalue by multimode reader.No false-positive results were observed for the other related bacteria used to evaluate assay specificity.The detection limit of the assay was up to 1.2CFU/mL.The intra-assay coefficient of variation was between 16.53%and17.97%.The inter-assay coefficient of variation was between 16.33% and 22.10%.The assay provides a new technical way for rapid detecting and preventing infection caused by Salmonella.

关 键 词：沙门菌 invA基因 夹心DNA杂交 

分 类 号：S852.612[农业科学—基础兽医学]