猪O型口蹄疫病毒抗体化学发光酶联免疫检测方法的建立  

Establishment of chemiluminescent enzyme immunoassay for detecting antibodies against foot-and-mouth disease virus serotype O in swine

作　　者：崔辰[1,2] 黄立纲 李晶[1] 邹兴启[4] 朱元源[4] 谢磊[5] 赵启祖[4] 杨利敏[1] 刘文军[1] Chen Cui Ligang Huang Jing Li Xingqi Zou Yuanyuan Zhu Lei Xie Qizu Zhao Limin Yang and Wenjun Liu(CAS Key Laboratory of Pathogenic Microbiology and Immunology, Institute of Microbiology, Chinese Academy of Sciences, Beijing 100101, China University of Chinese Academy of Sciences, Beijing 100049, China China Institute of Veterinary Drug Control, Beijing 100081, China Department of Clinical Laboratory, First Hospital of Tsinghua University, Beijing 100016, China Beijing Animal Health Inspection Institute, Beijing 100044, China)

机构地区：[1]中国科学院微生物研究所病原微生物与免疫学重点实验室,北京100101 [2]中国科学院大学,北京100049 [3]清华大学第一附属医院检验科,北京100016 [4]中国兽医药品监察所,北京100081 [5]北京市动物卫生监督所,北京100044

出　　处：《生物工程学报》2016年第11期1519-1530,共12页Chinese Journal of Biotechnology

基　　金：中国科学院重点部署项目(No.KSZD-EW-Z-005-001)资助~~

摘　　要：表达并纯化猪O型口蹄疫病毒(FMDV)VP1重组蛋白作为检测抗原,建立了一种快速检测猪O型口蹄疫病毒抗体的化学发光酶联免疫(CLEIA)检测方法。建立的VP1-CLEIA方法特异性为100%,板内变异系数在1.10%–6.70%之间,板间变异系数在0.66%–4.80%之间,具有较好的特异性和重复性,且灵敏度高于ELISA方法。通过对山东、辽宁、河北地区采集的250份临床血清的检测表明,该方法与间接ELISA试剂盒的符合率为93.50%,与液相阻断ELISA试剂盒的符合率为94.00%,表明本次建立的VP1-CLEIA检测方法可以用于猪O型FMDV感染或疫苗免疫后抗体水平检测。Recombinant structural protein VP1 of foot-and-mouth disease virus serotype O was expressed in Escherichia coli and then purified using Nickel affinity chromatography. A chemiluminescent enzyme immunoassay （CLEIA） method was established using the purified recombinant protein as coating antigen to detect antibody of foot-and-mouth disease virus serotype O in swine. The specificity of VP1-CLEIA method is 100%. The coefficients of variation in the plate and between plates are 1.10%-6.70% and 0.66%-4.80%, respectively. Comparing with the commercial indirect ELISA kit or liquid phase block ELISA kit, the calculated coincidence rate is 93.50% or 94.00%. The high specificity and stability suggested this detection method can be used to monitor the antibody level of foot-and-mouth disease virus serotype O in swine.

关 键 词：O型口蹄疫病毒 VP1蛋白 化学发光酶联免疫 

分 类 号：S852.65[农业科学—基础兽医学]