鸡毒支原体胶体金免疫层析试纸条的研制和初步应用  

Development and Application of Colloidal Gold Immunochromatographic Strip for Rapid Detection of Mycoplsma gallisepticum

作　　者：宁官保[1] 刘国莉 张鼎[3] 李宏全[1] 马海利[1] 高荣琨[1] 郝卫芳 高文伟[1] 赵宇军[1] 高诗敏[1] 李建慧[1] 李桂兰[1] 闫芳[1] 田文霞[1] 

机构地区：[1]山西农业大学动物科技学院,山西太谷030801 [2]介休市劳动和社会保障局,山西介休032000 [3]华中农业大学动物医学院,湖北武汉430070 [4]太原市动物疫病预防控制中心,山西太原030024

出　　处：《动物医学进展》2015年第5期25-28,共4页Progress In Veterinary Medicine

基　　金：山西省自然科学基金项目(2014011028-1;2009011043-1);山西省科技攻关项目(20130311027-3);国家自然科学基金项目(31072179)

摘　　要：为研制一种用于快速检测鸡毒支原体(MG)的胶体金免疫层析试纸条,采用直径20nm的胶体金颗粒标记纯化的MG多克隆抗体,硝酸纤维素膜检测线和质检线分别喷加纯化的MG多克隆抗体和兔抗鸡IgG抗体,制作胶体金试纸条。试纸条检测MG阳性显示两条红色反应条带;阴性为一条红色反应带。本试纸条可检测到1∶1 280倍稀释的MG抗原,即颜色变化单位为8×104 CCU/mL,具有高度的灵敏性。用试纸条检测衣原体、滑液支原体、猪肺炎支原体、鸡大肠埃希菌和鸡白痢沙门菌等均呈阴性,表明该试纸条具有较好的特异性。对试纸条检测呈MG阳性的38只病鸡进行MG培养验证,所得结果一致;同时对MG培养检测的另外95份病鸡样品进行试纸条测试比对,符合率为96.84%,证明该试纸条具有很高的准确性。该胶体金免疫层析试纸条检测MG操作简单,灵敏度、准确度、特异性等均符合要求,可以用于养殖场批量MG的检测。This study aimed to develop a colloidal gold immunochromatographic strip specific for Mycoplsma gallisepticum （MG） detection, a 20nm-diameter colloidal gold was selected to mark the purified MG anbi- body,which was then wrapped with nitrocellulose membrane. Another purified MG antibody and rabbit- anti-chicken IgG antibody was also connected to nitrocellulose membrane as test line and control line re- spectively. Two red reaction stripes both on the test and control line showed up when the finished strip was justified with the standard MG antigens while only one red line appeared on the control line in the neg- ative samples. The strips can detect the diluted MG antigens up to 1 ：1 280 （8 × 104 CCU/mL）, indicating a satisfied sensitivity. We also justified the qualified specificity of the strip by testing Chlamydia, Myco- plsma synoviae, Mycoplasma hyopneumoniae, chicken Escherichia coli and Salmonella, all of which pres- ented negative results. To evaluate the accuracy of the strip, we verified the 70 positive samples and 60 negative samples tested with strips using laboratory MG cultivation and plat agglutination methods. Re- suits showed that the strip had a total coincidence of 98.92% with laboratory MG cultivation way, and 96.97% with the plat agglutination method. In conclusion, the developed colloidal gold immunochromato- graphic strip had qualified requirements of specificity, sensitivity and accuracy, and was recommended to extensively apply in the clinic MG disease diagnosis.

关 键 词：鸡毒支原体 胶体金 免疫层析 试纸条 

分 类 号：S852.62[农业科学—基础兽医学]