牛乳中致病性金黄色葡萄球菌快速检测胶体金免疫层析方法研究  

Establishment of immunochromatographic method for rapid detection of pathogenic Staphylococcus aureus in milk

作　　者：布日额[1,2] 任晓峰[3] 吴金花[1,2] 锡林高娃[1,2] 王学理[2,4] 刘燕[1,2] 孙立杰[1,2] 刘洋[4] 

机构地区：[1]内蒙古民族大学生命科学学院,内蒙古通辽028043 [2]内蒙古民族大学乳源性致病菌研究所,内蒙古通辽028043 [3]东北农业大学动物医学院,黑龙江哈尔滨150030 [4]内蒙古民族大学动物科技学院,内蒙古通辽028043

出　　处：《中国预防兽医学报》2013年第11期907-911,共5页Chinese Journal of Preventive Veterinary Medicine

基　　金：内蒙古自治区科技厅科技创新引导计划项目(20111802;2012年;2013年)

摘　　要：为建立快速检测牛乳中致病性金黄色葡萄球菌(S.aureus)的胶体金免疫层析方法,本研究将鼠抗S.aureus IgG进行胶体金标记,并喷涂于玻璃纤维上制备金标垫,分别以兔抗S.aureus nEBPS重组蛋白IgG和羊抗鼠IgG作为检测线和质控线,组装制备快速检测牛乳中致病性S.aureus胶体金免疫层析试纸条。结果表明:试纸条对致病性S.aureus具有高度特异性,与沙门氏菌、大肠杆菌、布氏杆菌无交叉反应;试纸条最低检出1.0×105cfu/mL的S.aureus。试纸条批内及批间检测试验均具有良好的重复性。试纸条与S.aureus分离鉴定法符合率为90.0%、与PCR试剂盒符合率为83.3%。表明本研究建立的牛乳中致病性S.aureus免疫层析检测方法具有特异、敏感、稳定、操作简单、快捷等特点,适合于牛乳中致病性S.aureus的现场快速检测。To establish a rapid assay for detection of Staphylococcus aureus in milk, an immunochromatograpic strip was developed by blotted the rabbit anti-nEBPS protein on the membrane as test line, the rabbit anti-mouse IgG as the control line and mouse anti-S.aureus IgG labled with colloidal gold as a detection antibody. The results show the test strip was specific for pathogenic S.aureus detection in milk, but no cross-reaction with Salmonella, E.coli, Brucella in milk. The test strip was capable of detecting 1.0xl05 cfu/mL of S.aureus. The specificity and sensitivity of the test strip were reproducibility. The coincidence rate of the test strip was 90.0% with S.aureus isolation and identification method and 83.33% with PCR detection kit, respectively. These results demonstrated that the established immunochromatographic strip was higher specificity, sensiticity and stability for pathogenic S.aureus detection in milk, which was suitable for the on-site application.

关 键 词：牛乳 致病性金黄色葡萄球菌 弹性纤维结合蛋白 胶体金免疫层析试纸条 

分 类 号：S852.61[农业科学—基础兽医学]