非洲马瘟病毒分型RT-PCR检测方法的研究  

Study on Development of Serotype-specific RT-PCR Technology for Detection of AHSV

作　　者：赵文华 杨仕标 王金萍 李富祥 

机构地区：[1]农业部热带亚热带动物病毒学重点开放实验室/国家外来动物疫病诊断实验室/云南省畜牧兽医科学院,昆明650224

出　　处：《中国农学通报》2012年第2期43-47,共5页Chinese Agricultural Science Bulletin

基　　金：公益性行业(农业)科研专项"非洲猪瘟等重要外来动物疫病综合防控技术研究"(200903037);云南省农业科技创新工程项目"云南省规模化养殖场主要动物疫病防控关键技术研究与应用"(2008LA019)

摘　　要：为验证所引进9个血清型的非洲马瘟病毒,并同时对其分型鉴定方法有所探索,根据参考文献设计位于AHSV基因组L2片段的型特异性分型引物,用RT-PCR及测序方法进行实验研究。抽提所引进9种血清型的RNA,经RT-PCR扩增,各引物均有特异性扩增,经进一步的核苷酸序列测定及分析,证实所扩增片段均为非洲马瘟病毒L2基因相应位置片段;同时对AHSV分型RT-PCR检测方法进行了改进优化研究。实验证实分型引物对各RNA均有特异扩增,所引进9个血清型为正确血清型,并建立了适合实验室条件的AHSV分型鉴定方法。This study aimed to confirm the imported 9 serotypes of AHSV and have some explore and discovery on the differentiating method for AHSV. In the current study specific primers for differentiating each of the nine serotypes of AHSV were designed based on the L2 genome segment according to the reported references. RT-PCR method and sequencing were used. The RNA samples extracted from inactivated vaccine virus of the above 9 serotypes all could produce specific products amplified with the specific primers through RT-PCR method, and the amplification conditions were modified and optimized. The products amplified from the AHSV RNA had been sequenced and blasted through the internet. Conclusively, the developed RT-PCR method in this study is able to be employed as a technique for differentiating the 9 serotypes of AHSV, and we also confirmed the imported 9 serotypes of RNA.

关 键 词：非洲马瘟病毒 L2基因组片段 血清型 RT-PCR 

分 类 号：S855.3[农业科学—临床兽医学]