蛋鸡J亚群禽白血病病毒SD1009分离株的分离鉴定及全基因组序列分析

Isolation and identification molecular characterization of a subgroup J avian leukosis virus isolate SD1009

为分析我国J亚群禽白血病病毒(ALV-J)蛋鸡分离株的进化关系,本研究将山东省某鸡场采集的蛋鸡病料样品接种DF-1细胞系,利用ELISA群特异性抗原检测以及亚群特异性间接免疫荧光方法,分离鉴定得到一株ALV-J,命名为SD1009,并对其进行全基因组测序,将该序列与其他ALV-J代表性病毒株序列进行比较。结果表明:SD1009分离株的gag和pol基因相对保守,与各参考病毒株的同源性为95%～99%,env基因的同源性为91%～95%;在其5'UTR中出现了连续19 bp的插入突变,与TW-3577、SDAU09C3、JS09GY6、JS09GY3蛋鸡分离株的5'UTR基本一致,提示19 bp的插入现象可能是近年来蛋鸡ALV-J的进化趋势;此外,其3'UTR的rTM和DR区出现部分缺失现象,该缺失部分也可能与ALV-J进化相关。

A isolate of subgroup J avian leukosis virus （ALV-J）, designed SD1009, was isolated from layer chicken in Shandong province and identified by PCR, ELISA and indirect immunofluorescence assay （IFA）. The viral genome of this isolate was sequenced and compared with reference ALV-J strains in GenBank. The results show that the gag and pol of the isolate were relatively conservative, the sequence identity was ranged from 95% to 99% for gag and pol genes, and from 91% to 95% for env gene comparing with reference strains. However, a 19 bp insertion mutation in 5＇ UTR of SD1009 was similar to TW-3577, SDAU09C3, JS09GY6 and JS09GY3 strains from layer chicken. Besides, partial deletion of the DR and rTM region in 3＇UTR of SD1009 isolate was also identified in some recent isolated ALV-J. In conclution, those mutations might suggest evolution trend of ALV-J in layer chicken.