地方品系鸡中一株A亚群鸡白血病病毒的分离和鉴定

ISOLATION AND IDENTIFICATION OF SUBGROUP A AVIAN LEUKOSIS VIRUS FROM CHICKENS OF A LOCAL BREED

将种蛋孵化到9~11d,分别制备成鸡胚成纤维细胞（chicken fibroblast cells,CEF）培养,再将其细胞上清接种对内源性白血病病毒（avian leukosis virus,ALV）有抵抗力的DF1细胞,从山东某地方品系种蛋中分离到一株外源性ALV,SDAU09E1。用PCR扩增其囊膜蛋白基因（env）并隆测序后,将其gp85序列与已发表的各亚群ALV比较分析表明,该毒株与A亚群6个毒株同源性最高,为89.1%~90.9%,而与已发表的鸡的A、C、D、E亚群ALV的gp85的同源性仅在73.2%~87.9%之间,与目前国内最常见的J亚群的gp85的同源性更是低至30.3%~32.4%。这是我国地方品系鸡群中第一次分离和鉴定出ALV-A及其gp85基因。

The fertilized eggs were collected from a local breed chicken flock and incubated for 9-11 days,chicken fibroblast cells（CEF） were prepared from these embryos one by one and incubated for 7-9 days.The supernatant was collected from each CEF culture and inoculated into DF1 cells.The inoculated DF1 cell cultures were incubated for another 9 days and tested for avian leukosis virus（ALV） group specific p27 antigen in ELISA.A exogenous ALV,SDAU09E1,was identified.By PCR amplification of the gp85gene.Comparison of gp85 amino acid sequences between SDAU09E1 and other ALV strains of different subgroups indicated that SDAU09E1 had the highest homology with 6 reference strains of subgroup A（in the range of 89.1%-90.9%）,but the homology with other subgroups B,C,D and E was only in the range of 73.2%-87.9%.In addition,th homology between SDAU09E1 and several strains of subgroup J,most common in China,was as low as 30.3%-32.4%.This is the first report on isolation and identification of ALV subgroup A from local breed chickens in China.