1株诱发血管瘤的ALV-J的分离鉴定及其囊膜基因的进化分析

doi:10.16431/j.cnki.1671-7236.2023.11.024

摘要/Abstract

摘要： 【目的】了解湖北某黑羽蛋鸡场J亚群禽白血病（J-avian leukosis）的来源及其囊膜基因进化趋势，为湖北地区禽白血病流行病学调查提供资料。【方法】运用病理学、ELISA和Multi-PCR方法对疑似血管瘤型J亚群禽白血病病毒（J-Avian leucosis virus，ALV-J）病例进行实验室诊断，制备病毒液接种DF-1细胞进行病毒分离及间接免疫荧光试验（indirect immunofluorescence assay，IFA）检测，并通过DNAStar等软件分析该毒株囊膜蛋白编码基因。【结果】病例呈现典型血管瘤病变，p27抗原检测呈阳性，Multi-PCR出现ALV-J特异性条带，IFA结果显示特异性荧光，表明分离到1株ALV-J，命名为HB2021017。分离株HB2021017囊膜蛋白ENV、膜表面糖蛋白SU、跨膜蛋白TM与所引用的ALV-J毒株中的髓细胞瘤型毒株、髓细胞瘤和血管瘤混合型毒株、血管瘤型的ALV-J毒株的氨基酸序列相似性逐步升高。系统进化树分析显示，分离株HB2021017的env基因与中国地方品种鸡群中分离的诱发血管瘤的ALV-J毒株亲缘关系最近，处于同一分支；而与其他鸡群分离诱发血管瘤或髓细胞瘤的ALV-J毒株亲缘关系较远，处于不同的分支。【结论】本研究确诊并分离到1株诱发血管瘤的ALV-J（HB2021017），本土地方品种鸡群中分离的诱导血管瘤的ALV-J毒株囊膜基因已经形成了相对独立的进化分支。

关键词: 血管瘤型; J亚群禽白血病毒(ALV-J); 囊膜基因; 克隆; 进化

Abstract: 【Objective】 This study was aimed to investigate the origin of J-avian leukosis of a black-finned egg farm in Hubei and the evolutionary trend of its envelope gene, and provide information for the epidemiological investigation of avian leukosis in Hubei.【Method】 Pathological examination, ELISA and Multi-PCR were used for laboratory diagnosis of suspected cases of hemangioma J-Avian leucosis virus (ALV-J).Viral fluid was prepared and inoculated with DF-1 cells for virus isolation and indirect immunofluorescence (IFA) detection, and the gene encoding the envelope protein of this strain was analyzed by DNAStar and other softwares.【Result】 The case presented typical hemangioma typical symptomatic lesions, the p27 antigen test was positive, the Multi-PCR results showed ALV-J specific bands, the IFA results showed specific fluorescence, which indicated that 1 strain of ALV was isolated successfully, named HB2021017.The amino acid similarity were increased progressively between the envelope membrane protein ENV, membrane surface glycoprotein SU, transmembrane protein TM of the isolate HB2021017 and the corresponding proteins of the cited ALV-J strains of myeloblastoma, mixed myeloblastoma and hemangioma, and hemangioma-type ALV-J strains.Phylogenetic tree analysis showed that the env gene of the isolate HB2021017 was most closely related to the hemangioma-inducing ALV-J strain isolated from local breed chickens in China, and was in the same branch;While it was more distantly related to the hemangioma-inducing or myeloblastoma-inducing ALV-J strain isolated from other chickens, and was in different branches.【Conclution】 A strain of hemangioma-inducing ALV-J, named HB2021017, was isolated.The envelope genes of the hemangioma-inducing ALV-J strain isolated from local breed chickens had formed a relatively independent evolutionary branch.

Key words: hemangioma type; J-Avian leucosis virus (ALV-J); envelope gene; cloning; evolution

中图分类号:

S852.65⁺9.3

李佑君, 伍澜, 杨雨欣, 顾玉芳, 方春, 刘晶, 梁雄燕, 杨玉莹. 1株诱发血管瘤的ALV-J的分离鉴定及其囊膜基因的进化分析[J]. 中国畜牧兽医, 2023, 50(11): 4557-4567.

LI Youjun, WU Lan, YANG Yuxin, GU Yufang, FANG Chun, LIU Jing, LIANG Xiongyan, YANG Yuying. Isolation and Identification of a Hemangioma-inducing ALV-J Strain and Evolution Analysis of Envelope Genes[J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(11): 4557-4567.

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