野鸭源禽网状内皮组织增生症病毒的分离与鉴定  

Isolation and Identification of Reticuloendotheliosis Virus in Wild Birds

作　　者：姜莉莉[1,2,3] 祁小乐[1] 高玉龙[1] 邓小芸[2] 柴洪亮[2] 张礼洲[1] 贠炳岭[1] 秦立廷[1] 王永强[1] 高宏雷[1] 王笑梅[1] 华育平[2] 

机构地区：[1]中国农业科学院哈尔滨兽医研究所,兽医生物技术国家重点实验室,禽传染病研究室,哈尔滨150001 [2]东北林业大学野生动物资源学院,哈尔滨150040 [3]辽宁医学院畜牧兽医学院,锦州121001

出　　处：《畜牧兽医学报》2013年第8期1277-1282,共6页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：公益性行业(农业)科研专项经费资助(201203055);现代农业肉鸡产业技术体系建设(nycytx-42-G3-01)

摘　　要：利用扩增禽网状内皮组织增生症病毒(REV)长末端重复序列(LTR)的特异性引物作为检测引物,对采集于吉林省某地健康野鸭的脏器样品进行PCR检测。阳性样品经DF1培养增殖,进行病毒的分离,经PCR和间接免疫荧光(IFA)方法鉴定,确定分离到2株REV,分别来自于针尾鸭和绿头鸭,将分离病毒分别命名为DBYR1101和DBYR1102。克隆2个病毒株的主要保护性抗原基因gp90,并进行序列分析。结果显示,2个分离株gp90基因氨基酸相似性为99.5%;DBYR1101 gp90基因与REV 3个型的代表株170A、SNV和CSV的氨基酸相似性分别为95.7%、94.2%和98.2%;DBYR1102 gp90基因与REV 3个型的代表株170A、SNV和CSV的氨基酸相似性分别为95.2%、93.7%和97.7%;与中国早期南方分离株HA9901的氨基酸序列相似性分别为94.7%和94.2%;与中国北方分离株氨基酸序列相似性为99.2%～100%。核苷酸遗传进化分析表明,2个野鸟源REV分离株与现有的东北分离株亲缘关系较近,趋于形成1个北方分离群;与SNV株亲缘关系最远;与170A株亲缘关系较远;与美国和中国台湾地区的某些分离株相似性较高。该研究首次自野鸭体内分离到REV,丰富了REV流行病学资料,同时提醒我们重视野生鸟类迁徙在疾病传播中所扮演的角色,为深入研究REV致病机制、免疫抑制机制等奠定基础。The specific primer pairs for amplification of long terminal repeat sequences(LTR) of Reticuloendotheliosis virus(REV) was used in the PCR reaction to amplify the specific nucleic acid sequence of healthy wild birds collected in Jilin province.Positive samples were inoculated on the DF1 cells for virus reproduction.With identification of indirect immunofluorescence assay(IFA),PCR assay,two REV strains were isolated,one from the pintail(Coturnix cotu),another from mallard(Anas platyrhynchos).The two isolates were named DBYR1101 and DBYR1102,respectively.The gp90 gene encoding the most important protective protein was cloned and sequenced.Sequence analysis showed that the gp90 gene based amino acid homology between the two isolates is 99.5%.And they are more identical to the northeast China isolates(the amino acid homology of 99.2% to 99.9%) than to the early Chinese REV isolates HA9901(94.7% and 94.2%).The two isolates in this study and the northeast China isolates tend to group together in their own distinct phylogenetic clade.The gp90 amino acid homology between DBYR1101 and 3 subtype of REV170A,SNV and CSV was 95.7%,94% and 98.2% respectively,while DBYR1102 was 95.2%,93.7% and 97.7%.And the gp90 gene between the two isolates and the early southern isolate HA9901 were 94.7% and 94.2%.In addition,the 2 REV strains have a high identity with some REV strains in US and Taiwan,which classified as subtype Ⅲ.This is the first study to investigate the status of wild birds infected with REV,and the results of this paper will not only expands the epidemiological data of REV,but also remind us pay more attention to the role of wild bird migration in the spread of diseases.Meanwhile,it also makes a basis for further researching the pathogenic and immunosuppressive mechanisms.

关 键 词：野鸟 禽网状内皮组织增生病病毒 分离 鉴定 

分 类 号：S852.659.3[农业科学—基础兽医学]