半巢式RT-PCR快速检测犊牛轮状病毒方法的建立  

Rapid detection of bovine rotavirus with semi-nested RT-PCR assay

作　　者：栾婧婧[1] 杨少华[1] 张维军[2] 高运东[1] 仲跻峰[1] 赵宏坤[1] 

机构地区：[1]山东农科院奶牛研究中心,济南250100 [2]山东农业大学动物科技学院,泰安271018

出　　处：《中国人兽共患病学报》2006年第7期671-673,共3页Chinese Journal of Zoonoses

基　　金：山东三○工程项目(2003-3009);国家"十五"科技攻关奶业专项(2002BA518A16)

摘　　要：目的根据牛轮状病毒的VP7基因设计了一对可以检测牛轮状病毒G6血清型的引物,成功地检测了牛轮状病毒G6血清型。使用该方法检测牛病毒性腹泻-粘膜病病毒,检测结果为阴性。敏感性试验表明本实验方法可以检测10-4稀释的样品。通过对23份临床样本的检测证实,PCR方法比聚丙烯酰胺凝胶电泳和病毒的分离试验灵敏度高,可以快速诊断轮状病毒病,并且能够同时鉴定轮状病毒主要血清型。A pair of primers was designed according to the VP7 gene of bovine rotavirus with semi-nested RT-PCR assay for detecting the bovine rotavirus G6 serotype, and it was used to detect this type of virus successfully ,but the detection for viruses of bovine viral diarrhea and mucosa diseases showed negative results. Sensitivity testing indicated that this method could detect the presence of virus at a 10^-4 dilution of samples. Meanwhile, through the detection of virus in 23 clinical specimens in- eluding 2 duodenum samples , it was demonstrated that the sensitivity of this method was higher than those of PAGE assay or virus isolation. It is evident that this method can be used as a rapid method to detect viruses and can identify the main serotype of rotavirus.

关 键 词：牛轮状病毒 RT-PCR 

分 类 号：R373.2[医药卫生—病原生物学]