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非洲马瘟病毒分型RT-PCR检测方法的研究
引用本文:赵文华 杨仕标 王金萍 李富祥. 非洲马瘟病毒分型RT-PCR检测方法的研究[J]. 中国农学通报, 2012, 28(2): 43-47. DOI: 10.11924/j.issn.1000-6850.2011-2668
作者姓名:赵文华 杨仕标 王金萍 李富祥
作者单位: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
基金项目: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。

关 键 词:水稻  水稻  重组自交群体  化感作用  数量性状位点  
收稿时间:2011-09-18
修稿时间:2011-10-10

Study on Development of Serotype-specific RT-PCR Technology for Detection of AHSV
Zhao Wenhua , Yang Shibiao , Wang Jinping , Li Fuxiang. Study on Development of Serotype-specific RT-PCR Technology for Detection of AHSV[J]. Chinese Agricultural Science Bulletin, 2012, 28(2): 43-47. DOI: 10.11924/j.issn.1000-6850.2011-2668
Authors:Zhao Wenhua    Yang Shibiao    Wang Jinping    Li Fuxiang
Affiliation:(Laboratory of Tropical and Subtropical Animal Virology of MOA /National Exotic Animal Disease Diagnostic Laboratory/ Yunnan Academy of Animal Husbandry and Veterinary Medicine, Kunming 650224)
Abstract:
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.
Keywords:African horse sickness virus (AHSV)  L2 genome segment  serotype  RT-PCR
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