猪圆环病毒2d亚型黑龙江株的分离鉴定与遗传进化分析  

Identification and genetic variation analyses of porcine circovirus subtype 2d isolated from Heilongjiang Province

作　　者：马明爽 周娜娜 马茹梦 李月 王晓娜 李佳璇[1,2] 姜艳平 崔文[1,2] 李一经 唐丽杰[1,2] MA Ming-shuang;ZHOU Na-na;MA Ru-meng;LI Yue;WANG Xiao-na;LI Jia-xuan;JIANG Yan-ping;CUI Wen;LI Yi-jing;TANG Li-jie(College of Veterinary Medicine,Northeast Agricultural University,Harbin 150030,China;Heilongjiang Provincial Key Laboratory of Animal Disease Control and Formulation,Harbin 150030,China)

机构地区：[1]东北农业大学动物医学学院,黑龙江哈尔滨150030 [2]黑龙江省动物疾病防控与制剂创制重点实验室,黑龙江哈尔滨150030

出　　处：《中国兽医科学》2023年第10期1262-1269,共8页Chinese Veterinary Science

基　　金：国家重点研发计划项目(2017YFD0501100)。

摘　　要：为了鉴定黑龙江省某猪场疑似猪圆环病毒2型(PCV2)感染死亡的猪只病料,采集疑似PCV2感染的组织并对病毒进行分离培养,通过PCR、免疫荧光、免疫电镜对分离的病毒进行鉴定。对阳性样品进行全基因组扩增,测序后与GenBank上其他不同国家和地区来源的24株PCV的同源性、系统发育树和重组特性进行分析。结果显示,该毒株能够感染PK-15细胞,与藻红蛋白标记的PCV2 Cap蛋白单克隆抗体结合产生特异性荧光,电镜下可见约20 nm的病毒颗粒。分离得到的毒株基因型为PCV2d,命名为LJ0616株,其基因组全长1767 bp,序列上传至GenBank的登录号为ON500676。通过对其核苷酸同源性和重组特性分析,LJ0616株PCV2d与其他21株PCV2毒株的核苷酸序列同源性为55.2%~99.8%,且并未发现重组现象。本研究结果为黑龙江省PCV2的流行病学提供了基础数据,也为该病原的进一步研究奠定了基础。In order to diagnose pigs suspected of having died of porcine circovirus type 2(PCV2)in a pig farm in Heilongjiang Province,tissues suspected of PCV2 infection were collected,and the virus was isolated and identified.The isolated virus was identified by PCR,immunofluorescence,and immunoelectron microscopy.The whole genome of the positive sample was amplified and sequenced,and the homology,phylogenetic tree,and recombination characteristics of 24 PCV from other countries and regions on GenBank were analyzed.The results showed that the virus strain could infect PK-15 cells and bind to the phycoerythrin labeled PCV2 Cap protein monoclonal antibody to produce specific fluorescence.Under electron microscopy,virus particles of about 20 nm were visible.The genotype of the isolated strain is PCV2d,named LJ0616 strain,with a total genome length of 1769 bp.The sequence was uploaded to Gen Bank under the registration number ON500676.Through analysis of its nucleotide homology and recombination characteristics,the nucleotide sequence homology of LJ0616 strain PCV2d and 21 other PCV2 strains was55.2%—99.8%,and no recombination was found.The results of this study provide basic data for the epidemiology of PCV2 in Heilongjiang Province and lay a foundation for further research on the pathogen.

关 键 词：猪圆环病毒2d亚型 分离与鉴定 遗传进化分析 

分 类 号：S852.659.2[农业科学—基础兽医学]