猪伪狂犬病病毒和猪繁殖与呼吸综合征病毒抗体同步检测的液相芯片技术  

Simultaneous Detection of Antibodies against Porcine Pseudorabies Virus and Porcine Reproductive and Respiratory Syndrome Virus with Liquid Chip Technology

作　　者：肖丽 丛锋[1] 刘向楠[1] 李秀珍 饶丹[1] 练月晓 黄碧洪 张钰[1] 黄韧[1] 郭鹏举[1] 陈梅丽[1] XIAO Li;CONG Feng;LIU Xiang-nan;LI Xiu-zhen;RAO Dan;LIAN Yue-xiao;HUANG Bi-hong;ZHANG Yu;HUANG Ren;GUO Peng-ju;CHEN Mei-li(Guangdong Laboratory Animal Monitoring Institute ,Guangdong Key Laboratory of Laboratory Animals ,Guangzhou ,Guangdong , 510663 ,China)

机构地区：[1]广东省实验动物监测所/广东省实验动物重点实验室,广东广州510663

出　　处：《动物医学进展》2018年第4期32-35,共4页Progress In Veterinary Medicine

基　　金：广东省科技计划项目(2015B070701003;2017A070702001);广州市科技计划项目(201707010440;2016201604030059)

摘　　要：为了建立猪伪狂犬病毒抗体和猪繁殖与呼吸障碍症病毒抗体的同步检测技术,通过蛋白抗原偶联微球,建立单重和双重液相芯片检测方法,并对蛋白抗原偶联微球量进行优化,验证该方法的特异性,同时用此方法与ELISA试剂盒血清检测方法进行灵敏度比较。结果证明,猪繁殖与呼吸综合征病毒N蛋白和猪伪狂犬病毒GE蛋白的抗原最佳偶联量分别为每2.5×10~5个微球偶联8μg和2.5μg;特异性试验结果显示,这两个蛋白抗原偶联的微球与猪其他病原的抗体无交叉反应,说明特异性良好;与ELISA方法比较发现,针对同份猪血清样品,液相芯片法的猪伪狂犬病毒和猪繁殖与呼吸综合征病毒抗体的最高血清检出稀释倍数比ELISA法分别高66倍和32倍。说明成功建立了一种同步检测猪伪狂犬病毒和猪繁殖与呼吸综合征病毒抗体的液相芯片方法。To establish a serological method for the simultaneous detection of porcine pseudorabies virus antibody and porcine reproductive and respiratory disease virus antibody and to provide new technical sup- port for the simultaneous detection of these two swine virus antibodies, antigens were couplied with micro- spheres for establishing single and double liquid chip detection methods, and optimizing the amount of pro- tein antigen coupling microspheres,verifying the specificity of the method.The serum detection sensitivity of method was compared with ELISA kit. The results showed that the optimum coupling amounts of antigen of porcine reproductive and respiratory disease virus N protein and PRV GE protein were 8 μg and 2.5 μg per 2.5×10^5 microspheres respectively. The result of specific experiment showed that two protein antigen-conjugated microspheres has no cross-reactivity with other pig pathogen antibodies, indicating good specificity; compared with the ELISA method,the result showed that for the same pig serum samples, the limits of serum dilutions detected with antibodies were 32 and 66 fold higher respectively. In conclusion, this study successfully established a method for simultaneous detection of antibodies of porcine pseudorabies virus and porcine reproductive and respiratory disease virus.

关 键 词：液相芯片 猪伪狂犬病病毒 猪繁殖与呼吸综合征病毒 抗体 

分 类 号：S852.659.1[农业科学—基础兽医学] S854.43[农业科学—兽医学]