家畜戊型肝炎病毒诊断方法的建立及应用

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()RF2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 Gen-Bank,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.