猪伪狂犬病血清抗体gB-ELISA检测方法的建立  

Establishment of the Indirect ELISA for the Detection of Viral Glycoprotein B Antibodies against Swine Pseudorabies

作　　者：罗飞[1,2] 李宇琴 陈义平[1] 周洁[1,2] 

机构地区：[1]中国动物卫生与流行病学中心诊断液研究室,山东青岛226032 [2]扬州大学兽医学院,江苏扬州225009 [3]湖南九鼎科技(集团)有限公司,湖南岳阳414000

出　　处：《中国兽药杂志》2011年第7期10-13,共4页Chinese Journal of Veterinary Drug

基　　金："十一五"国家科技支撑项目(2006BAD6A13)

摘　　要：用纯化的猪伪狂犬病病毒gB重组蛋白为抗原,建立了检测猪伪狂犬病血清抗体的gB-ELISA方法。最佳反应条件为:抗原包被浓度为3.15μg/mL,待检血清稀释度为1∶40。该方法对猪圆环病毒病、猪瘟、猪细小病毒病、猪繁殖与呼吸综合征(猪蓝耳病)、猪乙型脑炎、猪布氏杆菌病5种疾病阳性血清和SPF猪阴性血清检测呈阴性反应。批间、批内试验变异系数均不超过8%。用该方法与HerdChek ELISA试剂盒同时对119份血清进行了平行检测,其相对敏感性、特异性和符合率分别为:75%、80.7%和79%。试验结果表明:猪伪狂犬病血清抗体gB-ELISA检测方法具有较高的敏感性和特异性,且重复性好,可用于猪伪狂犬病毒血清抗体检测。The gB-ELISA was established by using the purified recombinant protein as antigen for detecting PRV antibodies in pig sera.All reaction conditions of this gB-ELISA were explored.The optimal coating concentration of antigen is 3.15 μg/mL,the serum sample was diluted to 1∶40.The results are negative when the antiserum against PCV,PPV,HCV,JEV and PRRS were detected by this gB-ELISA.The CV between and within batches of detection were less than 8%.119 pig sera samples were simultaneously detected by this indirect gB-ELISA and HerdChek gB ELISA kit.The relative sensitivity and specificity of indirect gB-ELISA established in the study were 75% and 80.7%.The coincidence rate between these two ELISA was 79%.All results showed that this assay had a good sensitivity,specificity and repeatability,it could be used for detection of PRV antibody in pig sera.

关 键 词：猪伪狂犬病毒 抗体 gB-ELISA 

分 类 号：S852.65[农业科学—基础兽医学]