检测猪乙型脑炎抗体胶体金免疫层析法的建立与初步应用  

The GICA method for detection of porcine Japanese B encephalitis antibody

作　　者：田纯见[1] 贾坤[2] 林志雄[1] 何晓明[2] 罗琼[1] 罗长保[1] 鱼海琼[1] 陈茹[1] 彭运平 吴晓薇[1] 

机构地区：[1]广东出入境检验检疫局检验检疫技术中心,广东广州510623 [2]华南农业大学,广东广州510642 [3]广州万孚生物技术有限公司,广东广州510530

出　　处：《广东畜牧兽医科技》2013年第2期21-25,共5页Guangdong Journal of Animal and Veterinary Science

基　　金：广东出入境检验检疫局科研项目(2001GDK11);国家质检总局科研项目(2006IK004)

摘　　要：利用蔗糖密度梯度超速离心浓缩纯化vero细胞培养SA14-14-2株猪乙脑病毒灭活抗原,进行胶体金点样和配比优化,制备GICA试纸条。结果最优浓缩样品配比构成是PBS 1.32 mL、灭活病毒液24 mL、30%蔗糖2.6 mL、55%蔗糖6.6 mL;用于胶体金偶联的抗原浓度约2.80 mg/mL。试纸条能明显区别阴阳性血清,阳性血清稀释1:256能显示条带,优化后灵敏度达到1:1024。与猪繁殖与呼吸综合征、猪瘟、Ⅱ型圆环病毒病阳性血清不发生交叉反应。用于检测2群免疫母猪抗体阳性率分别为86%和100%,说明疫苗免疫效果较好;检测未经免疫肉猪出现阳性结果,分别达到38%和24%,说明存在病毒隐形感染。Inactivated antigens of JEV SA14-14-2 strain cultivated in vero cells were purified by sucrose density gradient ultracentrifugation. After spotting tests of colloidal gold and matching optimization, GICA strip was prepared. Results showed that the optimal ingredient formulation ratio is PBS 1.32 mL, inactivated virus solution 24 mL, 2.6 mL of 30% sucrose, 55% sucrose 6.6 mL. The concentration of inactivated virus solution was 2.80mg/mL. The strip can significantly distinguish between positive and negative serum. The positive serum sample after 1：256 dilution can display distinct bands. The sensitivity is up to 1：1024 after optimization. The positive sera anti PRRSV, CSFV and circovirus II had no cross reaction. Using GIGA strips to detect sera from two groups of sows immunized with JEV vaccine, the positive rates were 86% and 100% respectively, showed good immune effect of vaccine, sera from pigs without JEV immunization the positive results were 38% and 24% respectively, indicating the presence of wild virus infection.

关 键 词：乙型脑炎 胶体金免疫层析 超速离心 抗体 

分 类 号：S852.43[农业科学—基础兽医学]