猪Ⅱ型圆环病毒浙江株的分离及全基因组结构分析  

Isolation and Genomic Structure of Porcine Circovirus Type Ⅱ in Zhejiang

作　　者：陈庆新[1,2] 周继勇[1] 叶菊秀[1] 申会刚[1] 陈婷飞[1] 

机构地区：[1]浙江大学动物预防医学研究所 [2]浙江大学生命科学学院杭州310029

出　　处：《微生物学报》2004年第4期526-529,共4页Acta Microbiologica Sinica

基　　金：浙江省重大科技攻关项目 ( 2 0 0 3C12 0 12 ) ;国家自然科学基金 ( 3 0 3 70 0 5 2 )~~

摘　　要：用PCR方法对从浙江省猪场收集的、以淋巴结肿大和生长迟缓为特征的患猪腹股沟淋巴结进行PCV2检测。将检测为阳性的病料研磨、离心、过滤除菌后 ,接种PCV freePK 1 5细胞进行病毒分离 ,分离获得 3株病毒 ,命名为HZ0 2 0 1、HZ0 2 0 2、NB0 30 1。PK 1 5细胞增殖所得病毒离心纯化后 ,在电镜下可观察到直径约为 1 7～ 2 0nm ,呈二十面体对称的病毒粒子。以组织和病毒的细胞DNA为模板进行PCV2的全基因扩增 ,测序结果显示 ,3株病毒分离物全基因序列均由 1 76 7bp组成 ,直接来自病料与细胞分离毒株的核苷酸同源性为 1 0 0 %。基因组内含有 1 1个ORF。通过比较发现 ,分离毒株与PCV2参考株的同源性介于 94 2 %～ 99 7%之间 ;与PCV1参考株的同源性为77 2 %～ 77 9%。Using specific primer of PCV type Ⅱ, the nucleotide fragment was amplified by PCR from the inguinal lymph nodes collected from pig farms in Zhejiang province and characterized with enlarged lymph nodes and progressive weight loss. Positive samples detected with PCV2 type specific primer by PCR were homogenized with a mortar in MEM. The suspension was clarified and filtered. The suspension was mixed with PCV free PK 15 cells, and then incubated subsequently at 37℃ with 5% CO 2 in an incubator. Three isolates of porcine circovirus type 2 were isolated and named as HZ0201, HZ0202, and NB0301. Purified virus is a 17nm in diameter and icosahedral virion in transmission electron microscope. The complete genome of three virus isolates is composed of 1767 nucleotides containing 11 ORFs. Nucleotide comparison of the virus isolates adapted in PK15 cells revealed 100% identity to that origin from the lymphoid tissue of the diseased pigs. Genome of the virus isolates shared 94 2%～99 7% homology with PCV2 referrence strains from GenBank, 77 2%～77 9% homology with PCV1.

关 键 词：猪圆环病毒2型 形态 基因结构 分离 PCV2 浙江 

分 类 号：S852.65[农业科学—基础兽医学]