七彩山鸡F亚群禽白血病病毒的分离与鉴定

为了解广东某七彩山鸡种禽场禽白血病流行情况,通过接种DF-1细胞、ELISA抗原检测、聚合酶链式反应(PCR)、囊膜基因克隆测序等方法分离并鉴定出两株ALV-F,分别命名为FGD1801和FGD1802。为进一步了解分离株遗传进化特点,利用PCR方法扩增出env基因并测序,同时与各亚群参考毒株的gp85核苷酸序列进行对比。结果显示:这两个分离株env基因gp85片段长度都为1080 bp,预计编码360个氨基酸,FGD1801和FGD1802分离株gp85基因核苷酸序列的相似性为92.8%,与A、B、E、J和K亚群共26株ALV参考毒株相似性在47.0%~64.5%之间,而与F亚群参考株的相似性在92.0%~92.5%,显著高于其他亚群,且位于同一进化分支上。研究表明,从七彩山鸡分离的FGD1801和FGD1802属于ALV-F,是我国华南地区新发现的ALV亚群。

Isolation and Identification of Avian Leukosis Virus Subgroup F from Colorful Pheasant

To investigate the prevalence of colorful pheasant breeder farm infected by avian leukosis virus in Guangdong province of China,two strains of ALV-F designed as FGD1801 and FGD1802 were isolated and identified by inoculating plasma into DF-1 cells,antigen detection by ELISA,polymerase chain reaction(PCR),and membrane gene cloning and sequencing.In order to further understand the genetic evolution characteristics of isolates,the env genes of ALV-F isolates were compared with reference strains of other subgroup ALV.The results showed that the gp85 fragments of the two ALV-F isolates were 1080 bp in length and were expected to encode 360 amino acids.The nucleotide sequence similarity of gp85 among FGD1801,FGD1802 and the ALV-F reference strain was 92.0%to92.8%,while the similarity of gp85 gene with those of 26 reference strains in subgroups A,B,E,J and K were ranged from 47.0%to 64.5%.All the ALV-F isolates were located in the same evolutionary branch,greatly different from other ALV subgroups.The results indicated that the strains FGD1801 and FGD1802 isolated from colorful pheasants should belong to ALV-F,which was a newly discovered ALV subgroup in Southern China.