绵羊肺腺瘤病的病理学及RT-PCR诊断  

The pathological and RT-PCR diagnosis of sheep pulmonary adenomatosis

作　　者：梁化春[1] 齐景伟[1] 刘淑英[1] 徐萌杰[1] 

机构地区：[1]内蒙古农业大学动物科学与医学学院,内蒙古呼和浩特010018

出　　处：《中国预防兽医学报》2009年第6期443-447,共5页Chinese Journal of Preventive Veterinary Medicine

基　　金：国家自然科学基金(30560108);国家自然科学基金(30760078)

摘　　要：对呼和浩特市四子王旗某种羊场的疑似绵羊肺腺瘤病的4只病羊,通过临床诊断、病理组织学观察,PCR、RT-PCR和半巢式PCR技术对疑似病羊进行检测诊断。结果显示:(1)"小推车试验"时有鼻液流出;(2)剖检时可见肺肿大实变,切开肺脏时可见大量泡沫状液体从切面渗出;(3)病理组织学观察肺脏有大小不一的腺瘤灶,而且肺泡上皮细胞呈乳头状增生;(4)根据GenBank中登录的绵羊肺腺瘤病毒全序列(AF105220),设计合成3对特异性引物和3条巢式引物,经RT-PCR、PCR及半巢式PCR法扩增并测序,利用RT-PCR和PCR分别扩增出与预期大小一致的条带U3(175bp)、env(225bp)、gag(300bp),序列分析表明与引起该病的绵羊肺腺瘤病毒序列同源性较高,分别达97.2%、96.5%、94.4%。应用半巢式引物扩增片段分别为U3(133bp)、env(178bp)、gag(275bp)。经过以上方法确诊该羊场的疑似病例为绵阳肺腺瘤病。In order to diagnose the 4 sheep suspected sheep pulmonary adenomatosis （SPA）, the sheep in Huhhot were studied by clinical diagnosis, microscopic observation, PCR, RT-PCR and hemi-nested PCR. The result showed that： （1） The nasal fluid flow when ＂wheel-barrow test＂ ; （2） The lung was larger than normal, the surface showed many gray nodus, a lot of cystose fluid was effused from cut; （3） In pathohistological observetion, many adenoid nodus were in the lung, the alveolar epithelial cell accrem- entied like corpora mammillaria. The macrophage cells were full in alveolar space. （4） According to Jaagsiekte sheep retrovirus sequence （AF105220） in the GenBank, three pair of primers and three pair of hemi-nested primers were designed, the result of PCR and RT-PCR showed that the amplified fragments were U3 （175 bp）; env （225 bp） and gag （300 bp） respectively. the sequence analysis showed that homology was 97.2 %, 96.5 %, 94.4 % respectively. The semi-nested amplified fragments were U3 （133 bp）, env （178 bp）, gag （275 bp）. So the suspected sheep were diagnosed SPA by the above methods.

关 键 词：绵羊肺腺瘤病 绵羊肺腺瘤病毒 病理学 诊断 

分 类 号：S852.166[农业科学—基础兽医学]