1株E亚群禽白血病病毒的分离鉴定及遗传进化分析

doi:10.11843/j.issn.0366-6964.2022.09.025

摘要/Abstract

摘要： E亚群禽白血病病毒（ALV-E）是指存在于鸡染色体中的内源性逆转录病毒基因组DNA或片段。具有转录活性的ALV-E既会对鸡的生产性能（体重和产蛋率）产生负面影响，又能从抗体水平干扰对外源性ALV的鉴别诊断。为对黑龙江省某鸡场内一禽白血病病毒RT-PCR阳性病料进行病毒分离鉴定及分析其基因组特征和遗传进化情况，通过分子生物学、病毒形态学及全基因组序列测定方法对病毒培养物进行鉴定和分析，结果显示，该分离株可在CEF细胞盲传至第9代，电镜下可观察到近似球形、直径约为80 nm，并具有囊膜和纤突结构的病毒粒子，将其命名为HLJE2020株。序列分析结果显示，其全基因组序列中的gag和pol基因相对保守，LTR和env基因与ALV-E同属一个进化分支，而gp85基因则与ALV-E和ALV-B均具有较高相似性，遗传进化分析显示在ALV-E和ALV-B间出现一个单独的分支，结合RDPv.4和SimPlot软件分析结果，推测该毒株gp85基因可能存在E亚群AF229株与B亚群SDAU09C1株的重组现象。本研究为了解禽白血病病毒基因组遗传演化情况提供数据资料，并为ALV的防控提供参考和依据。

关键词: 禽白血病病毒, 分离鉴定, 基因组分析, 重组E亚群毒株

Abstract: Avian leukemia (leukosis) virus subgroup E (ALV-E) refers to endogenous retroviral genomic DNA or fragments present in chicken chromosomes. ALV-E with transcriptional activity will not only have a negative impact on chicken production performance (body weight and egg production), but also interfere with the differential diagnosis of exogenous ALV at the antibody level. In order to isolate and identify an avian leukosis virus RT-PCR positive disease material in a chicken farm in Heilongjiang Province and analyze its genome characteristics and genetic evolution, the virus culture was identified and analyzed by molecular biology, virus morphology and whole genome sequencing. Results showed that the virus could be stably subcultured to the nine generation in CEF cells. Virus particles with approximate spherical shape, diameter of about 80 nm, capsule and fiber structure could be observed using electron microscopy. It was named HLJE2020 strain. Sequence analysis showed that gag and pol genes in the whole genome sequences were relatively conserved. LTR and env genes belong to the same evolutionary branch with ALV-E, while gp85 gene had high homology with ALV-E and ALV-B. Genetic evolution analysis combined with the analysis results of RDPv.4 and SimPlot software showed that there was a separate branch between ALV-E and ALV-B. It is speculated that the gp85 gene of this virus may be recombined with subgroup E AF229 and subgroup B SDAU09C1. This study can provide data for understanding the genetic evolution of avian leukemia virus genome, and provide references and basis for ALV prevention and control.

Key words: avian leukemia virus, isolation and identification, genome analysis, strain of recombinant subgroup E

中图分类号:

S852.659.3

王萌, 张森豪, 张柳, 李佳璇, 王晓娜, 崔文, 姜艳平, 周晗, 王丽, 乔薪瑗, 李一经, 唐丽杰. 1株E亚群禽白血病病毒的分离鉴定及遗传进化分析[J]. 畜牧兽医学报, 2022, 53(9): 3107-3120.

WANG Meng, ZHANG Senhao, ZHANG Liu, LI Jiaxuan, WANG Xiaona, CUI Wen, JIANG Yanping, ZHOU Han, WANG Li, QIAO Xinyuan, LI Yijing, TANG Lijie. Isolation,Identification and Genetic Evolution Analysis of a Subgroup E Avian Leukemia Virus[J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(9): 3107-3120.

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