口蹄疫O型、A型和AsiaⅠ型病毒RT-PCR分型方法的建立  

Establishment of RT-PCR Genotyping Method for Foot-and-Mouth Disease Virus Type O,A and Asia Ⅰ

作　　者：上官陶[1] 王洪梅[1] 宋玲玲[1] 武建明[1] 于力[2] 何洪彬[1] 

机构地区：[1]山东省农业科学院奶牛研究中心,山东济南250100 [2]中国农业科学院哈尔滨兽医研究所

出　　处：《现代农业科技》2013年第14期249-250,254,共3页Modern Agricultural Science and Technology

基　　金：泰山学者海外特聘专家(tshw20100417);现代农业(奶牛)产业技术体系岗位科学家(Cars-37);国家自然科学基金项目(31272686);济南市高效院所自主创新计划(201004027);济南市高效院所自主创新计划(201202059);山东省自然科学基金(ZR2010CQ029)

摘　　要：该研究旨在建立一种快速、灵敏及准确的口蹄疫病毒(FMDV)鉴定与分型的RT-PCR检测技术平台,为临床中有效准确地检测口蹄疫病毒提供有力的支持。根据NCBI中公布的FMDV序列(GenBank:JF749861.1),设计FMDV通用检测引物FP1/FP2;根据NCBI中公布的FMDV O型、A型和Asia Ⅰ型序列,经过序列分析比对后,分别设计FMDV分型引物FO1/FO2、FA1/FA2和FAS1/FAS2。提取FMDVRNA后,利用随机引物扩增得到FMDV全cDNA,利用设计的引物进行PCR扩增及PCR反应条件的优化。结果表明:该研究建立的RT-PCR检测方法具有很好的特异性、敏感性和可重复性。成功建立了FMDV O型、A型和Asia Ⅰ型检测及分型方法,为口蹄疫疾病的临床检测。To establish a rapid ,sensitive and accurate foot-and-mouth disease virus（FMDV）identification and genotyping of RT-PCR detection technology,and provide strong support for the effective and accurate detection of foot-and-mouth disease virus in clinical,we designed the FMDV general detection primer FP1/FP2 according to the the NCBI published in FMDV sequence （GenBank ：JF749861.1 ） ,and designed the FMDV genotyping primers FO 1/FO2, FA 1/FA2 and FAS1/FAS2 according to the FMDV type O, type A and type Asia I sequence from NCBI after sequence analysis.After extraction of the FMDV RNA,we got the FMDV cDNA using random primers, and used cDNA as the template for PCR amplification and optimized the PCR reaction conditions.The results showed that RT-PCR genotyping methods established in this study had a good specificity,sensitivity and reproducibility.The results indicated that the RT-PCR genotyping method for foot-and-mouth disease virus Type O,A and Asia I had been successfully established,and laid the foundation for the clinical detection epidemiological investigation and targeted vaccine preparation of FMD disease in clinical research.

关 键 词：口蹄疫 RT-PCR O型 A型 AsiaⅠ型 

分 类 号：S855.3[农业科学—临床兽医学]