|
|||||
|
|
| 猪戊型肝炎病毒荧光定量PCR检测方法的建立及其分子流行病学 | |
| 引用本文： | 张晓峰,帅江冰,李爱云,方维焕,何永强,吴珊.猪戊型肝炎病毒荧光定量PCR检测方法的建立及其分子流行病学[J].中国兽医学报,2011,31(6). |
| 作者姓名： | 张晓峰 帅江冰 李爱云 方维焕 何永强 吴珊 |
| 作者单位： | 1. 浙江出入境检验检疫局,浙江杭州,310012 2. 浙江大学,医学院,附属妇产科医院,浙江杭州310006 3. 浙江大学,动物预防医学研究所,浙江杭州310029 |
| 基金项目： | 浙江省国境安全检验检疫科技创新服务平台资助项目(2006C17014); 浙江出入境检验检疫局科技基金资助项目(ZK200611) |
| 摘 要： | 对已发表的戊型肝炎病毒(HEV)不同基因型毒株全序列进行分析,针对ORF3的保守区设计引物和探针优化建立了HEV荧光RT-PCR检测体系。分析表明该反应体系具有良好的扩增效率、特异性和稳定性,且检测灵敏度比普通RT-PCR方法高10～100倍。利用所建立的荧光定量PCR方法对采集自浙江及其周边上海、江苏等地区规模化猪场的样品以及HEV抗体阳性人血清样品进行了HEV核酸检测。其中,77份人血清样本仅有2份为HEV核酸阳性(2.6%)。而猪粪便样品阳性率则高达20.5%(56/273),且所调查的猪场均存在HEV感染。进一步的分子流行特征分析表明,浙江及周边地区的猪HEV毒株多为基因Ⅳ型,与其他地区的Ⅳ型HEV毒株同源性较高(83.4%～89.5%),但它们之间也存在较多的变异位点。猪源HEV毒株PJ-1则与多数Ⅲ型HEV毒株同源性较高(88%),进化分析也表明其为基因Ⅲ型。人血清样品中检测到的2份HEV毒株Human-1与Human-2与本地区Ⅳ型猪源毒株在分子进化关系上具有很高的亲源性,它们分布于同一分支内,而不构成独立分支。以上分析结果表明浙江及其周边地区猪HEV感染较为广泛,且以基因Ⅳ型毒株为主,但也存在... |
| 关 键 词： | 戊型肝炎病毒 荧光定量PCR 基因型 分子进化 |
Establishment of real time PCR assay for hepatitis E virus from swine and its molecular epidemiology |
|
| ZHANG Xiao-feng,SHUAI Jiang-bing,LI Ai-yun,FANG Wei-huan,HE Yong-qiang,WU Shan.Establishment of real time PCR assay for hepatitis E virus from swine and its molecular epidemiology[J].Chinese Journal of Veterinary Science,2011,31(6). | |
| Authors: | ZHANG Xiao-feng SHUAI Jiang-bing LI Ai-yun FANG Wei-huan HE Yong-qiang WU Shan |
| Institution: | ZHANG Xiao-feng1,SHUAI Jiang-bing1,LI Ai-yun2,FANG Wei-huan3,HE Yong-qiang1,WU Shan1(1.Zhejiang Entry-Exit Inspection and Quarantine Bureau,Hangzhou 310012,China,2.Women's Hospital,School of Medicine,Zhejiang University,Hangzhou 310006,3.Zhejiang Provincial Key Laboratory of Preventive Veterinary Medicine,Hangzhou 310029,China) |
| Abstract: | A Taqman-based real time PCR assay was developed for detection of hepatitis E virus according to the ORF3 gene.The assay showed highly specificity and stability,as well as a 10-100 fold higher sensitivity as compaired with conventional RT-PCR.The carrier status of hepatitis E virus(HEV) in pig populations in different pig farms of Zhejiang province,as well as in HEV-positive human sera was examined by the assay.The results showed that only 2 as positive samples of 77 human sera were detected by real time PC... |
| Keywords: | hepatitis E virus real time PCR genotype phylogenetic analysis |
| 本文献已被 CNKI 万方数据 等数据库收录！ | |