黄羽鸡J亚群禽白血病病毒的分离及gp85基因分析

Isolation and gp85 Gene Analysis of Avian Leukosis Virus Subgroup J from Yellow Chicken

为了解禽白血病病毒在黄羽鸡中的流行情况,采用DF-1细胞接种、细胞培养上清P27抗原检测、PCR扩增,对送检疑似禽白血病的黄羽鸡病料进行了病毒分离,并对分离病毒gp85基因进行了测序和序列比较.结果表明,自黄羽鸡分离到2株J亚群禽白血病病毒（ALV-J）,分别命名为AHaq01和AHaq02,它们之间的核苷酸序列同源性为93.8％,与ALV-J参考毒株序列同源性为91.1％～99.8％,其中与ALV-J原型毒株HPRS-103的序列同源性分别为93.3％和98.1％,与分离株AHaq01和AHaq02同源性最高的分别是WN100404（95.0％）和SD09TA04 （99.8％）.进化分析进一步表明,2个分离株间的亲缘关系较远,为来源不同的ALV-J毒株.

In order to understand the epidemic situation of avian leukosis virus in the yellow chicken, two strains of avian leukosis virus subgroup J （ALV-J） were isolated from the yellow chicken by DF-1 cell cul- ture,detection of p27 antigen and RT-PCR, which were named as strains AHaq01 and AHaq02. The gp85 genes of the two isolated ALV-J strains were amplified by PCR, and were compared with the nucleotide se- quences of the reference ALV-J strains. The two isolated strains had 91.1%-99.8% sequence identities to the reference ALV-J strains, AHaq01 and AHaq02 have 93.3% and 98. 1% sequence identities to strain HPRS-103, respectively. AHaq01 had the highest homology with strain WN100404, AHaq02 had the highest homology with strain SD09TA04, and the homology of the two isolated strains was 93.8%. Phy- logenetic analysis based on gp85 gene showed the two isolated strains had distant genetic relationship,and had the closest linkage to WN100404 and SD09TA04, respectively.