检测犬源狂犬病毒中和抗体ELISA方法的建立  

Development of an ELISA for Detection of Neutralizing Antibodies Against Canine Rabies Virus

作　　者：傅秋玲[1] 郑佳琳[1] 林颖仪[1] 张莹[1] 阳佑天[1] 潘姣姣[1] 吴晓薇[1] 郭霄峰[1] 

机构地区：[1]华南农业大学兽医学院,广东广州510642

出　　处：《华南农业大学学报》2012年第4期561-565,共5页Journal of South China Agricultural University

基　　金：广东省省部产学研重点项目(2010A090200083);农业公益性行业专项(201103032)

摘　　要：为了有效监测犬免疫狂犬病疫苗后的保护效力,以狂犬病毒(Rabies virus,RV)糖蛋白的主要优势抗原表位区G3蛋白(RV G3)作为包被抗原,建立了一种检测狂犬病毒中和抗体效价的间接ELISA方法.通过优化反应条件,确定抗原最佳包被量为8 mg/L,血清的最佳稀释度为1∶100,酶标二抗的稀释度为1∶3 000.特异性试验表明,该抗原不与犬瘟热病毒、犬腺病毒、犬细小病毒阳性血清发生交叉反应;批内和批间重复性试验的平均变异系数都小于10%;敏感性达1∶1 280.此方法检测134份血清样品的结果与美国SYNBIOTICS试剂盒相比,总符合率达95.6%.To monitor the effectiveness of canine rabies vaccination, the indirect ELISA method had been developed to detect the neutralizing antibodies against rabies virus （RV） by using the main antigenic de- terminant of glycoprotein （ RV G3 ） as coating antigen. The optimum test conditions were as follows ： the optimal concentration of RV G3 coating the ELISA plate was 8 mg/L. The optimal concentration of serum samples and HRP-labeled goat anti-canine IgG was 1：100 and 1：3 000 respectively. RV G3 had no reac- tion with positive serum against canine distemper virus （ CDV）, canine adenovirus （CAV） or canine par- vovirus （CPV） by the specificity test. The coefficients of variation of intro-batch and inter-batch duplica- bility test were fewer than 10%. The sensitivity was 1：1 280. Compared with America SYNBIOTICS kit, the coincidence rate of indirect ELISA was 95.6%. Therefore, the indirect ELISA has good specificity, repeatability and sensitivity, which can be a good candidate for routine detection of neutralizing antibodies of canine rabies.

关 键 词：狂犬病毒 糖蛋白 犬源 中和抗体 间接ELISA 

分 类 号：S855[农业科学—临床兽医学]