屠宰猪肝脏中戊型肝炎病毒(HEV)的检测及组织病理学观察  

Detection of the Hepatitis E virus(HEV) and Its Histopathologic Observation in Liver of Slaughtered Swine

作　　者：王英华[1] 韦海涛 佘锐萍[1] 赵景义 李文贵[1] 李秀敏 刘利强[1] 韩德平[1] 王德成[1] 于品[1] 孙泉[1] 

机构地区：[1]中国农业大学动物医学院,北京100094 [2]北京畜牧兽医总站,北京100107

出　　处：《农业生物技术学报》2008年第3期373-379,共7页Journal of Agricultural Biotechnology

基　　金：国家自然科学基金资助项目(No.30771588)资助

摘　　要：从北京、河北和河南三地的屠宰场共采集了581头屠宰猪的肝脏。用抗-人戊型肝炎病毒(HEV)抗体做肝脏中HEV抗原的免疫组织化学染色,用针对HEVORF2区设计的通用引物进行巢式RT-PCR,结果发现,6个地区581头屠宰猪的肝脏HEV免疫组织化学阳性率都在90%以上。从114份肝脏样品中扩增到2条产物,与预期片段348bp大小基本一致,位于HEVORF2区,序列分析、比较显示,扩增出的2个片段(GenBank登录号为:EU326142和EU375565)同源性为99.2%,与HEV基因Ⅰ、Ⅱ、Ⅲ和Ⅳ型的同源性分别为74.9%～78.7%,73.9%～74.1%,74.6%～77%和81.2%～93.7%。用扩增的核苷酸片段绘制的基因进化树显示,2个分离株与HEV基因Ⅳ型AJ344171、AJ344172和AJ344183位于同一分支,属于HEV基因Ⅳ型。Five hundred and eighty one swine liver samples were collected from Slaughterhouse in Beijing, Henan and Hebei Provinces. Immunohistochemical stain and nested RT-PCR were carried out to analysis the current status of Hepatitis E virus （HEV） infection. Results showed that over 90% of the livers were positive for immunohistochemical stain. And 2 positive samples for HEV RNA were found from 114 liver samples detected by nested RT-PCR. Sequence comparison showed that 2 Beijing strains（GenBank accession No. EU326142 and EU375565） shared 99.2% homology within each other, and 74.9%～78.7%,73.9%～74.1%,74.6%～77% and 81.2%～93.7% to genotypes Ⅰ、Ⅱ、Ⅲand Ⅳ, respectively. Phylogenetic tree analyses further revealed that the two positive swine HEV isolates were closely related to HEV AJ344171, AJ344172 and AJ344183 isolates and belonged to genotype Ⅳ.

关 键 词：戊型肝炎病毒(HEV) 免疫组化 组织病理学 巢式RT—PCR 屠宰猪 

分 类 号：S185[农业科学—农业基础科学] S188