新疆绵羊肺腺瘤病理学诊断与全基因组序列分析  

Pathological Diagnoses and Whole-genome Sequence Analyses of the Jaagsiekte Sheep Retrovirus in Xinjiang,China

作　　者：杨素芳[1] 梁田[1] 赵庆亮[1] 张殿卿[1] 司俊强 张静[1] 杨霞[1] 盛金良[1] 

机构地区：[1]石河子大学动物科技学院,石河子832003

出　　处：《病毒学报》2015年第3期217-225,共9页Chinese Journal of Virology

基　　金：国家自然科学基金(30960276;31360589)

摘　　要：为了完成新疆绵羊肺腺瘤病毒诊断及全基因组序列测定,本研究采用兽医临床病理学和组织病理学观察疑似患病羊,并提取病毒悬液进行透射电镜观察,从患病绵羊的肺脏组织中提取基因RNA反转录为cDNA。参照GenBank中外源性绵羊肺腺瘤病毒美国株基因序列(AF105220)设计六对引物,对病料样品进行RT-PCR扩增和测序分析。结果表明,"小推车试验"时有鼻液流出,显微镜下观察,患病羊的肺脏有大小不一的腺瘤灶,且肺泡上皮细胞呈乳头状增生,肺泡腔内充满巨噬细胞,病灶中央的细胞核溶解。透射电镜观察到病毒颗粒直径大小约为88nm^125.4nm。RT-PCR扩增病毒基因组片段,获得完整的基因组序列全长7 456bp。与美国代表株(AF105220)和英国代表株(AF357971)BLAST在线比对核苷酸同源性结果分别为96%和95%,与内源性绵羊肺腺瘤病毒(enJSRV)内蒙株(DQ838493)和美国株(EF680300)的核苷酸同源性分别为89.8%和89.9%。获得的全基因组序列的TM区的氨基酸序列中具有外源性绵羊肺腺瘤病毒(exJSRV)特有的致病性"YXXM"序列。说明此基因序列具有致瘤作用,这是我国新疆首次报道的外源性绵羊肺腺瘤病毒的全基因组序列,为更深入的研究新疆绵羊肺腺瘤病毒的生物学特性和致病机理奠定基础。To carry out pathologic diagnoses and whole-genome sequence analyses of the Jaagsiekte sheep retrovirus（JSRV）in Xinjiang,China,we first observed sheep suspected to have the JSRV.Then,the extracted virus suspension was observed by transmission electron microscopy（TEM）.Total RNAs from lungs of JSRV-infected sheep were extracted and reverse-transcribed using a cDNA synthesis kit.Six pairs of primers were designed according to the exogenous reference virus strain（AF105220）.Reverse transcription-polymerase chain reaction was carried out from JSRV-infected tissue,and the whole genome of the JSRV sequenced.Our results showed：flow of nasal fluid（ ＂ wheelbarrow test＂）;different sizes of adenoma lesions in the lungs;papillary hyperplasia of alveolar epithelial cells;alveolar cavity filled with macrophages;dissolute nuclei in central lesions.TEM revealed JSRV particles with a diameter of 88 nm to125.4nm.The full-length of the viral genome sequence was 7456 bp.BLAST analyses showed nucleotide homology of 96%and 95%compared with that of the representative strain from the USA（AF105220）and UK（AF357971）.Nucleotide homology was 89.8% and 89.9% compared with the endogenous Jaagsiekte sheep retrovirus,Inner Mongolia strain（DQ838493）and USA strain（EF680300）.The specific pathogenic amino-acid sequence ＂ YXXM＂ was found in the TM district,similar to the exogenous JSRV：this gene has been reported to be oncogenic.This is the first report of the complete genomic sequence of the exogenous JSRV from Xinjiang,and could lay the foundation for study of the biological characteristics and pathogenic mechanisms of the pulmonary adenomatosis virus in sheep.

关 键 词：绵羊肺腺瘤病毒 病理组织学观察 RT-PCR扩增 序列分析 

分 类 号：S852.65[农业科学—基础兽医学]