摘要
根据戊型肝炎病毒(HEV)的4个基因亚型,寻找其保守序列ORF2,成功设计了一套以Ⅰ、Ⅳ型为主兼顾Ⅱ、Ⅲ型的通用简并引物,摸索条件,建立检测动物戊型肝炎病毒反转录套式聚合酶链反应(RT-nPCR)的方法,扩增片段为507 bp。该方法检测所需时间为5 h,灵敏度可达10 pg,特异性强、重复性好;经优化研制了动物戊型肝炎病毒RT-nPCR检测试剂盒,保存期半年以上,该试剂盒可用于动物粪便、胆汁HEV RNA的检测。
Based on the conserved nucleotides of the hepatitis E virus （HEV） genome sequences of four major genotypes, two pairs of degenerate primers were designed to amplify a fragment of 507bp within ORF2 by reverse transcription nested polymerase chain reaction （RT-nPCR）.The methed could be finished with in 5 hours and detect 10pg RNA with good specificity and repeatability. Through optimizateion, the established Kit can be kept for at least half a year used for the detecion of HEV RNA from animal feces and bile.
出处
《中国动物检疫》
CAS
2010年第11期34-35,共2页
China Animal Health Inspection
基金
上海市科技兴农重点攻关项目
沪农科攻字(2006)第5-7号
