5株猪圆环病毒2型全基因克隆和序列分析

Full Length Sequence Cloning and Sequence Analysis of 5 Porcine Circovirus Type 2 Isolates

为了解广东地区猪圆环病毒2型(PCV2)流行势态及毒株的基因变异情况,从广东珠三角地区疑似断奶仔猪多系统衰竭综合征(PMWS)患病死亡猪中采集淋巴结,将PCR鉴定为阳性的病料在PK-15细胞上进行增殖培养,成功分离到5株猪圆环病毒2型毒株,分别标记为GD-jm、GD-sz、GD-pz、GD-gj和GD-ss。再从细胞培养物中提取病毒基因组DNA,对5株PCV2分离株全基因组进行PCR扩增并克隆到pMD18-T Simple Vector进行测序,测序结果提交GenBank,登录号分别为JX912914、JX912915、JX945575、JX945576和JX945577。借助相关生物学软件作同源性分析,这5株PCV2基因组长度均为1767bp,其中3株为PCV2b亚型,2株为PCV2d亚型。

To understand the prevalent trend and genetic variations of porcine circovirus type 2（PCV2） from pig farms in Guangdong province,lymphonodi associated with postweaning multisystemic wasting syndrome were collected from pearl river delta in Guangdong province and then the disease materials that proved to be positive by polymerase chain reaction（PCR） method were cultured in PK-15 cells and 5 PCV2 strains were successfully isolated and marked as GD-jm,GD-sz,GD-pz,GD-gj and GD-ss,respectively.The viral genome DNA was extracted and the whole genome of 5 PCV2 isolates were amplified and inserted into pMD18-T Simple Vector.The results were submitted into GenBank and the accession numbers were JX912914,JX912915,JX945575,JX945576 and JX945577.Sequence homology analysis was conducted with the aid of biology software.We found that all of 5 isolates harbored a full length sequence of 1767 bp and 3 of which were designated into PCV2b genotype,the other two were designated into PCV2d genotype.