与鸡内皮血管瘤病例相关的禽白血病病毒K亚群分离及其gp85基因演化分析  

Isolation of ALV-K Associated with Endothelial Hemangioma in Chicken and Analysis of gp 85 Gene Evolution

作　　者：梁灿新 郑小雪 舒雪利 周婉怡 廖明[1] 曹伟胜 LIANG Canxin;ZHENG Xiaoxue;SHU Xueli;ZHOU Wanyi;LIAO Ming;CAO Weisheng(National and Local Joint Engineering Laboratory for Zoonoses Prevention and Control Agents,Key Laboratory of Veterinary Vaccine Creation of Ministry of Agriculture,Key Laboratory of Prevention and Control of Zoonotic Diseases of Guangdong Province,College of Veterinary Medicine,South China Agricultural University,Guangzhou 510642,China)

机构地区：[1]华南农业大学兽医学院,广东省动物源性人兽共患病预防与控制重点实验室,人兽共患病防控制剂国家地方联合工程实验室,农业部兽用疫苗创制重点实验室,广州510642

出　　处：《畜牧兽医学报》2024年第3期1127-1136,共10页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：广东省家禽产业技术体系疾病控制岗位专家(2022KJ128,2023KJ128);广东省重点领域研发计划(2020B020222001);国家肉鸡产业技术体系(CARS-41);2021年省乡村振兴战略专项资金(大专项+任务清单)项目(5500-F220137)。

摘　　要：本研究在华南地区某规模化种禽场调查中发现其部分花鸡品系头部鸡冠下缘及眼睑上缘部位出现白色、质地坚硬且形状不规则的肿块,为了解患病鸡的发病特征及致病因子而开展了实验室诊断及相关病原研究。结果表明该规模化养殖场主要发病鸡群为父母代花鸡,发病率约为2%,剖检无其他肉眼表观病变,病理组织学诊断为内皮血管瘤。RT-PCR检测肿瘤组织表明为K亚群禽白血病病毒(ALV-K)感染,无其他禽肿瘤性病毒感染。对患病鸡的血浆样品(32份)、肿瘤组织匀浆(6份)及脑组织匀浆(6份)进行病毒的分离鉴定,成功从脑组织样品中分离到4个ALV-K毒株(GXJL01~GXJL04)。使用特异性引物扩增GXJL01~GXJL04的env基因并测序,在与华南地区其他5个规模化种禽场中分离获得的7个ALV-K流行株的gp85基因的遗传演化分析中表明,GXJL01~GXJL04与日本的Km5844、Sp53等神经胶质瘤毒株(fowl glioma-inducing virus, FGV)及具有神经组织嗜性的ALV-K毒株JS15SG01同一进化分支上,与本研究其他7个ALV-K分离株的遗传进化关系较远。同时在针对ALV-K的gp85蛋白氨基酸位点突变分析中发现,GXJL01~GXJL04存在多个与FGV和JS15SG01参考株相似的氨基酸位点突变。本研究首次把地方黄羽鸡品种花鸡头部的肿块诊断为内皮血管瘤,且ALV-K为内皮血管瘤的主要相关病原,分离株GXJL01~GXJL04区别于现行华南地区ALV-K流行株且与FGV、JS15SG01等神经组织嗜性的毒株高度同源。In the study,we found that one type of tumors in yellow-feather chicken that look like a white,stiff,irregular shaped mass,growing up at the position between comb and eyelid.In order to identify the pathogenetic characteristics and causative agent of the disease,we have made a series of diagnoses and studies of the disease.Research has found that about 2%sick chicken in the laying hens,they have no other pathological changes other than the tumors.The histopathological diagnosis the pathological tissue was Hemangioendothelioma.RT-PCR tests on tumor tissue showed that the sick chickens were infected with the subgroup K avian leukosis virus(ALV-K).We have tried to isolate the viruses from 32 blood samples,6 tumor tissues and brain tissues of infected chicks.Finally,we successfully isolated 4 strains of ALV-K from brain tissues.The 4 strains of ALV-K were named as GXJL01-GXJL04.To better understand the molecular characterization of GXJL01-GXJL04.On the other hand,we have isolated another 7 strains of ALV-K from 5 large-scale chicken farms in South China,amplifying and sequencing their env genome.The phylogenetic tree of 11 strains of ALV-K was drawn by MEGA-X software based on gp85 amino acid sequence,indicating that GXJL01-GXJL04 were most similar to the JS15SG01 and fowl glioma-inducing virus(FGV)such as Km5844 and Sp53,and they were distantly related to the genetic evolution of the other 7 ALV-K isolates in this study.What’s more,in the analysis of gp85 amino acid point mutation,we have found that some amino acid sites are the same as fowl glioma-inducing virus and JS15SG01,while they are different from other 7 strains of ALV-K from south China.In the study,the tumors of the chicken was firstly diagnosed as Hemangioendothelioma,the associated pathogen was ALV-K,and the isolates GXJL01-GXJL04 were highly homologous to neurotropic strains like FGV,JS15SG01,but were different from the other circulating ALV-K in South China.

关 键 词：黄羽肉鸡 鸡内皮血管瘤 K亚群禽白血病病毒 遗传进化分析 

分 类 号：S855.3[农业科学—临床兽医学]