两株J亚型禽白血病病毒分离鉴定及全基因序列分析

doi:10.11843/j.issn.0366-6964.2025.12.035

摘要/Abstract

摘要： 为了探究禽白血病病毒(ALV)的进化趋势和分子特征，本研究从上海市某鸡场采集了1 600份血液样品，接种鸡成纤维细胞系(DF-1)分离病毒，通过PCR扩增gp85基因序列进行亚群分型，进一步通过间接免疫荧光试验(IFA)和多步生长曲线对其生物学特性进行鉴定，并将分离株和ALV各亚群毒株的编码区和非编码区进行序列比对和分子构象分析。结果显示，通过PCR和IFA鉴定本研究共分离到了6株ALV-J亚群毒株，其中ALV-J-345株和ALV-J-480株序列存在明显差异，且ALV-J-480株在DF-1细胞上比ALV-J-345株具有更强的复制能力，进一步比对两株分离株和ALV-J其他毒株的序列发现，两株分离株的gp85和gp37蛋白之间共存在46个差异氨基酸位点，且ALV-J-480株和其他ALV-J毒株的相似性更低，除此之外ALV-J-480株pol基因在第866个氨基酸位置提前产生一个终止密码子，使其缩短了8个氨基酸，在非编码区位置上ALV-J-345株在3'UTR的E(XSR)元件区缺失了7个核苷酸，而ALV-J-480株在5'UTR的第358~376位置处多插入了19 nt的核苷酸，这些位置上的序列差异，可能是导致ALV-J-480株具有更强复制能力的原因，这使得其在鸡群中有成为新的流行毒株的竞争优势。因此本研究提供了关于ALV-J分子特征的最新数据，这将有助于揭示ALV-J的进化趋势，并为制定相关的防控措施提供参考。

关键词: 禽白血病病毒, 分离鉴定, gp85, 序列分析, 基因变异

Abstract: To investigate the evolutionary trends and molecular characteristics of Avian leukosis virus (ALV), this study collected 1 600 blood samples from a poultry farm in Shanghai. Viruses were isolated by inoculating chicken fibroblast cells (DF-1), followed by subgroup identification through PCR amplification of the gp85 gene. Biological characterization was performed using indirect immunofluorescence assay (IFA) and multi-step growth curves. Coding and non-coding regions of the isolated strains were compared with representative ALV subgroups for sequence alignment and molecular conformation analysis. The results showed that six ALV-J strains were successfully isolated and identified via PCR and IFA. Notably, strains ALV-J-345 and ALV-J-480 exhibited significant sequence divergence, with strain ALV-J-480 demonstrating stronger replication efficiency in DF-1 cells than strain ALV-J-345. Further comparison with other ALV-J strains revealed 46 distinct amino acid variations in the gp85 and gp37 protein between the two isolates, and strain ALV-J-480 displayed lower homology to other ALV-J strains. Additionally, a premature termination codon at position 866 in the pol gene of strain ALV-J-480 truncated the protein by eight amino acids. In non-coding regions, strain ALV-J-345 harbored a 7-nucleotide deletion in the E(XSR) element region of the 3'UTR, whereas strain ALV-J-480 carried a 19 nt nucleotide insertion at positions 358-376 in the 5'UTR. These sequence variations likely contributed to the enhanced replication capacity of strain ALV-J-480, endowing it with a competitive advantage to emerge as a potential epidemic strain in poultry populations. Therefore, this study provides updated molecular insights into ALV-J, which will aid in elucidating its evolutionary dynamics and inform the development of targeted control strategies.

Key words: avian leukosis virus, isolation and identification, gp85, sequence analysis, genetic variation

中图分类号:

S852.659.3

夏琦琦, 桂亚萍, 刘健, 杨显超, 陈琦, 沈莉萍, 冯桂丹, 向蒙, 徐平, 唐聪圣, 卫龙兴, 洪云超, 石慧花, 王建, 赵洪进. 两株J亚型禽白血病病毒分离鉴定及全基因序列分析[J]. 畜牧兽医学报, 2025, 56(12): 6326-6338.

XIA Qiqi, GUI Yaping, LIU Jian, YANG Xianchao, CHEN Qi, SHEN Liping, FENG Guidan, XIANG Meng, XU Ping, TANG Congsheng, WEI Longxing, HONG Yunchao, SHI Huihua, WANG Jian, ZHAO Hongjin. Isolation, Identification, and Whole Gene Sequence Analysis of Two J Subtype Avian Leukemia Virus[J]. Acta Veterinaria et Zootechnica Sinica, 2025, 56(12): 6326-6338.

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