中国动物检疫2025，Vol.42Issue(3)：91-100,10.DOI:10.3969/j.issn.1005-944X.2025.03.015
猪圆环病毒3型阻断ELISA抗体检测方法的建立与应用
Establishment and Application of a Blocking ELISA for Porcine Circovirus Type 3
摘要
Abstract
In order to establish a blocking ELISA for porcine circovirus type 3(PCV3),PCV3 capsid(Cap)protein was expressed using the Escherichia coli expression system,2 strains of Cap protein monoclonal antibodies(1D8 and 3B5)were prepared,and dissociation constants(KD)of the monoclonal antibodies were measured;1D8 with high affinity was selected and marked with horseradish peroxidase(HRP),and a blocking ELISA for testing PCV3 was established based on Cap protein and the 1D8 marked with HRP.The results showed that the serum was determined to be positive for PCV3 antibodies when the present inhibition(PI)was≥29.3%,to be negative when the PI was＜23.4%,and to be suspicious when the PI was≥23.4%but＜29.3%.By the established method,no cross reaction with the antisera against PVC2,porcine reproductive and respiratory syndrome virus(PRRSV)and classical swine fever virus(CSFV)was observed;for the method,the sensitivity was 1:160,coefficients of variation for intra-and inter-batch repeated tests ranged from 0.87%to 6.64%and from 1.03%to 9.33%,respectively,and the coincidence rate with Western-blot assay was 95.5%.205 serum samples collected from some swine farms in Hebei Province from 2021 to 2024 were detected by the established method,it was found that the positivity rate of PCV3 antibodies was 28.8%(59/205),to be specific,30.0%,32.8%and 19.5%in piglets,fattening pigs and sows,respectively.In conclusion,the established method could be used for serologic diagnosis and epidemiologic investigation of PCV3 infection,indicating a great potential for future development and application.关键词
PCV3/Cap蛋白/单克隆抗体/阻断ELISA/应用Key words
PCV3/Cap protein/monoclonal antibody/blocking ELISA/application分类
农业科技引用本文复制引用
侯佳璐,任静,王亚文,袁晨,马天天,杨柳,李清艳,宋勤叶..猪圆环病毒3型阻断ELISA抗体检测方法的建立与应用[J].中国动物检疫,2025,42(3):91-100,10.基金项目
驻冀高校研究生与石家庄市企业合作项目(20240101) （20240101）
国家生猪技术创新中心项目(NCTIP-XD/B11) （NCTIP-XD/B11）
