猪圆环病毒2型特异性IgA间接ELISA检测方法的建立与初步应用  

Development and primary application of indirect ELISA method for detecting the IgA against porcine circovirus type 2

作　　者：刘雨丰[1] 马攀攀[1] 陈陆[1] 杨霞[1] 姚惠霞[1] 王川庆[1] 赵军[1] 

机构地区：[1]河南农业大学牧医工程学院,河南郑州450002

出　　处：《中国预防兽医学报》2012年第4期301-304,共4页Chinese Journal of Preventive Veterinary Medicine

基　　金：国家自然科学基金(31072116)

摘　　要：为检测猪圆环病毒2型(PCV2)特异性IgA抗体,本研究以纯化的PCV2 Cap蛋白作为包被抗原,以辣根过氧化物酶(HRP)标记的羊抗猪IgA为二抗,建立检测PCV2特异性IgA抗体的间接ELISA方法。确定最佳抗原包被浓度为200 ng/孔,HRP标记羊抗猪IgA的最佳稀释度为1∶30 000。所建立方法与抗猪瘟病毒等病原的抗体间无交叉反应。结果表明该方法具有较好的特异性,组内和组间重复性试验的变异系数介于0.12%～5.47%,具有较好的可重复性。该方法为进一步研究猪感染PCV2和PCV2疫苗免疫后特异性黏膜IgA水平及进行黏膜免疫评价提供了有效的检测方法。An indirect ELISA for detecting the IgA against porcine circovirus type 2（PCV2） was developed by using purified PCV2 Cap protein as coating antigen and HRP-conjugated goat anti-pig IgA as second antibody.Concentration of coating antigen and dilution of second antibody were optimized as 200 ng/well and 1∶30,000 respectively.There was no cross-reactivity with anti-HCV,PRRSV and other pathogen antibodies.The coefficient variation of intra-and inter-assay was ranged from 0.1233% to 5.4766%.The ELISA described in this study was proved to be a specific and rapid for the detection of PCV2 specific IgA.It was potential to be applied for both laboratorial and field detection of the PCV2 specific IgA induced by PCV2 infection or PCV2 vaccination and evaluating mucosal immune response.

关 键 词：猪圆环病毒2型 黏膜IgA 间接ELISA 

分 类 号：S855.3[农业科学—临床兽医学]