猪瘟病毒化学发光抗体检测方法的建立与应用  

Establishment and Application of Chemiluminescence Detection Method for Antibody against Classical Swine Fever Virus

作　　者：麻园 石正旺 罗俊聪 杨波[2] 王丽娟 万颖 宋锐 曹丽艳 周改静 田宏[2] 郑海学[2] 陈轶霞[1] MA Yuan;SHI Zhengwang;LUO Juncong;YANG Bo;WANG Lijuan;WAN Ying;SONG Rui;CAO Liyan;ZHOU Gaijing;TIAN Hong;ZHENG Haixue;CHEN Yixia(College of Life Science and Engineering,Northwest Minzu University,Lanzhou 730030,China;State Key Laboratory of Lanzhou Veterinary Research Institute,Chinese Academy of Agricultural Sciences,Lanzhou 730046,China)

机构地区：[1]西北民族大学生命科学与工程学院,兰州730030 [2]中国农业科学院兰州兽医研究所国家重点实验室,兰州730046

出　　处：《畜牧兽医学报》2021年第6期1744-1752,共9页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：西北民族大学中央高校基本科研业务项目(31920190027);非洲猪瘟科技应急防控专题(20190211);西北民族大学预防兽医学创新团队项目(110130143)。

摘　　要：旨在建立猪瘟病毒(CSFV)化学发光抗体检测方法,本研究以CSFV E2蛋白作为包被抗原,山羊抗猪IgG-HRP抗体作为酶标抗体,鲁米诺为底物溶液,优化检测方法,成功建立CSFV化学发光抗体检测方法。该方法能在室温20 min内完成对CSFV抗体血清特异性检测,灵敏度与商品化CSFV抗体检测试剂盒相当,且与A型口蹄疫病毒、O型口蹄疫病毒、猪繁殖与呼吸综合征病毒、塞内卡病毒、非洲猪瘟病毒抗体阳性血清均无交叉反应;批内变异系数为1.80%~6.88%,批间变异系数为1.11%~9.18%,重复性好。通过对152份田间猪血清样品的检测并与商品化CSFV抗体检测试剂盒检测结果进行比较,其Kappa值为0.929,具有高度的一致性。综上表明,本研究建立的CSFV化学发光抗体检测方法特异性强、灵敏性高、重复性好、简单快速,可应用于临床血清CSFV抗体的检测。The aim of this study was to establish a chemiluminescence detection method for antibody against classical swine fever virus (CSFV), CSFV E2 protein was employed as the antigen, which was probed and bound by goat anti-pig IgG-HRP antibody. The signal was quantified by adding Luminol as the substrate solution. Compared to the commercial CSFV antibody detection kit, this method is characterized by fast detection (20 min at room temperature), equivalent sensitivity and great specificity, as evidenced by no cross-reaction with positive serum samples of A and O serotype of foot-and-mouth disease virus, porcine reproductive and respiratory syndrome virus, senecavirus A and African swine fever virus. Meanwhile, this method demonstrated high repeatability with low intraassay coefficient of variation (CV) (1.80%-6.88%) and interassay CV (1.11%-9.18%). The Kappa value of 152 pig serum samples collected from the field was 0.929, which was highly consistent with the results of the commercial CSFV antibody detection kit. In conclusion, the CSFV chemiluminescence antibody detection method established in this study displayed strong specificity, high sensitivity and repeatability, which can be applied to the clinical detection of CSFV serum antibody.

关 键 词：猪瘟病毒 化学发光 抗体检测方法 

分 类 号：S852.651[农业科学—基础兽医学]