猪圆环病毒2型毒株的分离鉴定及其对小鼠的免疫原性分析  

Isolation,Identification and Immunogenicity Analysis of Porcine Circovirus Type 2 in Mice

作　　者：孙瑶 韩伟 赵辉 杨小蓉 常昊天 付秀花 吴珊珊 王贵华 Sun Yao;Han Wei;Zhao Hui;Yang Xiaorong;Chang Haotian;Fu Xiuhua;Wu Shanshan;Wang Guihua(China Animal Husbandry Industry Co.,Ltd.,Beijing 100070,China;Key Laboratory of Veterinary Biological Products and Chemical Drugs of Ministry of Agriculture and Rural Affairs,Beijing 100095,China;Beijing Veterinary Peptide Vaccine Design and Preparation Technology Center,Beijing 100095,China)

机构地区：[1]中牧实业股份有限公司,北京100070 [2]农业农村部兽用生物制品与化学药品重点实验室,北京100095 [3]北京市兽用多肽疫苗设计与制备工程技术中心,北京100095

出　　处：《中国动物检疫》2025年第1期103-110,共8页China Animal Health Inspection

摘　　要：为获得猪圆环病毒2型(PCV2)候选疫苗株,对疑似PCV2感染猪的淋巴结、脾脏等组织病料进行检测,对阳性病料进行PCV2分离纯化、鉴定、全基因测序和遗传进化分析,并制成PCV2灭活疫苗,进行小鼠免疫原性分析。结果显示;从PCV2阳性病料中成功分离得到1株PCV2毒株,病毒含量不低于10^(6.0) FAID_(50)/mL,间接免疫荧光试验可见绿色荧光;该毒株基因全长约1767 bp,全基因核酸序列与PCV2参考序列ON012565.1、KU960933.1同源性最高为99.9%,与PCV3参考序列PP566974.1同源性最低为45.9%,与多数PCV2参考毒株序列同源性在92%以上,经进化树分析属于PCV2d基因亚型;用该分离毒株制成灭活疫苗免疫小鼠后,测得特异性抗体,且效价较高。结果说明,该分离株具有良好的免疫原性,可作为PCV2疫苗研发的候选毒株。In order to obtain a candidate vaccine strain for porcine circovirus type 2(PCV2),lymph node,spleen and other tissue samples of pigs suspected of being infected with PCV2 were detected,followed by isolation,purification,identification,whole gene sequencing and genetic evolution analysis based on positive samples,to prepare PCV2 inactivated vaccines.Then immunogenicity analysis in mice was carried out.The results showed that one PCV2 strain was successfully isolated from the positive samples,and the virus titer was not less than 10^(6.0) FAID_(50)/mL,green fluorescence was observed through indirect immunofluorescence assay(IFA);the strain was about 1767 bp in its total gene length,and its whole gene nucleic acid sequence was max.99.9%identical to PCV2 reference sequences ON012565.1 and KU960933.1,min.45.9%to PCV3 reference sequence PP566974.1,and above 92%to most PCV2 reference sequences,falling into PCV2d gene subtype based on evolutionary tree analysis;specific antibodies were detected with high titer after vaccinating mice with inactivated vaccines made from the isolate.In conclusion,the isolate could be used as a candidate to develop PCV2 vaccines by virtue of its good immunogenicity.

关 键 词：猪圆环病毒2型 遗传变异 免疫原性分析 疫苗研发 

分 类 号：S851.3[农业科学—预防兽医学]