兔出血症病毒RT-PCR检测方法的建立及其临床应用  

Establishment and Clinical Application of RT-PCR Assay for Detection of Rabbit Hemorrhagic Disease Virus

作　　者：胡波[1] 魏后军[1] 王芳[1] 范志宇[1] 徐鹏[1] 徐为中[1] 薛家宾[1] 何孔旺[1] 

机构地区：[1]江苏省农业科学院兽医研究所农业部动物疫病诊断与免疫重点开放实验室国家兽用生物制品工程技术研究中心,南京210014

出　　处：《畜牧兽医学报》2010年第11期1442-1446,共5页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：农业部公益性行业科研专项(nyhyzx07-040);现代农业产业技术体系建设专项资金资助项目(nycytx-44);江苏省科技支撑计划--农业部分(BE2008372);江苏省农业科技自主创新资金项目(cx(08)107)

摘　　要：本研究旨在建立检测兔群鼻拭子中兔出血症病毒(RHDV)的RT-PCR方法,研究健康免疫兔群是否存在携带RHDV现象。根据GenBank上公布的RHDVvp60基因保守序列,设计了1对特异性引物,经cDNA的合成和PCR扩增,目的片段大小为591bp。结果表明该方法能检出最小RNA浓度为2.40ng·μL-1,敏感性为血凝试验(HA)的8×103倍。通过对自5个省采集的168份健康免疫兔鼻拭子样品进行检测,结果显示,阳性样品为22份,阳性率为13.09%。试验表明:RT-PCR方法能快速、敏感地从健康免疫兔鼻拭子样品中检出RHDV,提示健康免疫兔群存在携带RHDV的现象,该方法适合临床进行大规模病原学检测和兔群携带病毒的调查,具有良好的应用前景。The RT-PCR assay for detection of Rabbit hemorrhagic disease virus（RHDV） in nasal swab specimens collected from rabbits was developed for the research of virus carrying in healthy immunized rabbit.A pair of primers were designed according to the conserved sequences of vp60 from RHDV published in GenBank,and a 591 bp segment was amplified by RT-PCR.The least amount of RHDV could be detected was 2.40 ng·μL-1,and the sensitivity of the RT-PCR was 8×103 times higher than that of HA method.168 nasal swabs collected from 5 provinces was detected by the assay,the results showed that the positive samples was 22 and the positive rate was 13.09%.These results indicated that the phenomenon of virus carrying in healthy immunized rabbit and the developed RT-PCR might be a promising approach in clinical screening,detection and carrying research of RHDV in a large number of immunized rabbit nasal swabs.

关 键 词：兔出血症病毒 RT-PCR 鼻拭子 病毒携带 

分 类 号：S852.659.6[农业科学—基础兽医学]