Isolation and Identification of a Field Isolate of Marek's Disease Virus with Acute Oncogenicity
Isolation and Identification of a Field Isolate of Marek's Disease Virus with Acute Oncogenicity
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发表时间：
2007
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通讯作者：
Meilan Mo
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作者：
Meilan Mo
从接种CVI988/Rispens疫苗的商品三黄鸡群中，通过将鸭胚成纤维细胞(DEF)单层细胞与禽外周血淋巴细胞(PBL)共转染，从商品三黄鸡群中分离到一株MDV地方野毒株YL040920。通过空斑形成时间和空斑形状的比较，获得了9个分离株，证明该分离株具有较高的纯度。结果表明，该毒株具有MDV-1毒株的序列特征。用抗MEQ单抗3G12E6进行免疫荧光染色(FA)，发现该毒株具有MDV-1毒株的序列特征。用该毒株攻击夏燕雏鸡，在等电点21d左右出现明显的肿瘤，表明该毒株具有急性致瘤性。在所有器官中，心脏、肝脏和皮肤的肿瘤发生率最高。t检验结果表明，从死亡率和肿瘤发生率来看，普通系雏鸡显著高于MD抗性系雏鸡(P<0.05)，而HVT免疫组和未免疫组之间差异不显著(P>0.05)。建立了一种基于多重聚合酶链式反应的禽类肿瘤病鉴别诊断技术，对感染MDV-1的雏鸡和攻毒雏鸡肿瘤标本进行了DNA检测，所有被检测的样品均能观察到MDV-1强毒的特征图谱，但对网状内皮组织增殖症病毒(REV)和禽白血病病毒(ALV)的检测均为阴性。研究结果表明，YL040920分离株，已证实为MDV-1纯毒力病毒，不含CVI988病毒、网状内皮组织增生症病毒(REV)和禽白血病病毒(ALV)，具有引起急性肿瘤的能力，主要发生在心脏、肝脏和皮肤。
A local field isolate of MDV named YL040920 was isolated from a commercial Three-Yellow chicken flock,which had been vaccinated with CVI988/Rispens vaccine and experienced significant MD visceral tumors,by transfecting monolayer of duck embryo fibroblast(DEF) with the peripheral blood lymphocytes(PBLs) of the bird.Nine individual clones of the isolate obtained by plaque assay were identical in the time of plaque formation and shape of plaques that demonstrated the purity of the isolate.The oncogenicity-related gene meq of the isolate was amplified by polymerase chain reaction(PCR) technique,and then was sequenced and compared with that of the other reference strains of MDV-1.The isolate was found to have the sequence characteristics of virulent MDV-1.Specific fluorescent staining localized in the nuclei of the infected DEFs by an immunofluorescence assay(FA) using an anti-MEQ monoclonal antibody 3G12E6.Xiayan chicks were challenged with the isolate and obvious tumors were observed at about PI 21d,showing the acute oncogenicity of the isolate.And among all the organs,heart,liver and skin had the highest tumor occurrences.The results of t-test showed that in term of the mortality and tumor occurrence,chickens of general line were significantly higher than those of chickens of MD resistant line(P0.05),however there were no significant difference between the vaccinated and unvaccinated chickens with HVT(P0.05).A developed differential diagnosis technique based on a multiplex PCR for avian neoplastic diseases was used to detect the DNA prepared from the virus infected DEFs and tumor samples of the challenged birds,the characteristic patterns of electrophoresis of the virulent MDV-1 were observed from all the samples detected,but were negative for detection of reticuloendotheliosis virus(REV) and avian leucosis virus(ALV).The results of the study demonstrated that isolate YL040920,proved to be a pure virulent virus of MDV-1 and free of CVI988 virus,reticuloendotheliosis virus (REV) and avian leucosis virus(ALV),posscesses the ability to cause acute tumors which mainly ocurred in heart,liver and skin.