家畜戊型肝炎病毒诊断方法的建立及应用  

Establishment and application of RT-nPCR for the detection of hepatitis E virus in domestic animal

作　　者：苏乾莲 李斌 赵武 秦毅斌 梁家幸 肖爱欢[2] 段群棚 何颖 黄伟坚[2] 

机构地区：[1]广西壮族自治区兽医研究所,广西畜禽疫苗所技术重点实验室,南宁530001 [2]广西大学动物科学技术学院,南宁530005

出　　处：《中国人兽共患病学报》2011年第11期1051-1053,共3页Chinese Journal of Zoonoses

基　　金：广西基本科研业务费专项(桂科专项10-1桂科专项11-1桂兽研专项08-3)联合资助

摘　　要：目的建立检测家畜戊型肝炎病毒(HEV)反转录套式聚合酶链式反应(RT-nPCR)方法,并应用于广西地区家畜戊型肝炎感染调查。方法参考GenBank上4种基因型HEV核苷酸序列,应用生物软件Oligo6.0在ORF2保守区设计两对简并引物,扩增目的片段为436bp,进行了特异性、敏感性和重复性试验,并对采自广西地区猪、牛和羊的肝粪样本进行HEV检测。结果检测家畜粪便样本阳性率31.49%(285/905);家畜肝脏样本阳性率18.01%(49/272)。结论所建立的RT-nPCR方法适用于家畜肝粪样本HEV检测,临床检测应用表明广西家畜存在HEV感染。The reverse transcription nested polymerase chain reaction（RT-nPCR） was developed to detect hepatitis E virus（HEV）in domestic animal clinical samples in Guangxi.Referring to four genotypes nucleotide sequences of HEV in GenBank,two pairs of degenerate primers were designed by Oligo6.0 to amplify a fragment of 436bp within conservative regions of ORF2,and it＇s specificity,sensitivity and repeatability was evaluated.The RT-nPCR assay was specific to hepatitis E virus and had good sensitivity and repeatability.The 1177 domestic animal samples including 905 feces samples and 272 liver samples were detected by RT-nPCR and results showed that 31.49%（285/905）feces samples and 18.01%（49/272）liver samples were positive.The clinical detection result revealed that the investigated domestic animal had been infected HEV in Guangxi.

关 键 词：戊型肝炎病毒 反转录套式聚合酶链式反应 临床检测 

分 类 号：R373.2[医药卫生—病原生物学]