国内不同基因型Asia1型口蹄疫病毒RT-PCR检测方法的建立  

Development of RT-PCR Method for Determination of Foot-and-Mouth Disease Virus Type Asia 1 of Various Topotypes

作　　者：于长勇[1,2] 金宁一[1] 王春凤[2] 鲁会军[1] 胡博[1,3] 刘昊[1,4] 张丹[1,3] 牟伟锋[1,5] 丛艳昭[1,5] 谷长维[1,5] 常巧呈[1,5] 刘存霞[1,6] 颜雯[1,3] 叶明[1,5] 

机构地区：[1]军事医学科学院军事兽医研究所病毒室,长春130062 [2]吉林农业大学动物科技学院,长春130118 [3]吉林大学畜牧兽医学院,长春130062 [4]云南农业大学动物科技学院,昆明650201 [5]延边大学农学院动物医学系,吉林龙井133400 [6]黑龙江八一农垦大学动物科技学院,黑龙江大庆163319

出　　处：《中国生物制品学杂志》2009年第11期1132-1135,共4页Chinese Journal of Biologicals

基　　金：国家科技支撑项目(2007BAD55B05);国家"863"项目(2006AA10A204)

摘　　要：目的建立口蹄疫病毒(FMDV)Asia1型江苏/05谱系及新疆/03谱系毒株的二联RT-PCR检测方法。方法在比较大量国内外FMDV流行毒株序列的基础上,设计检测不同基因型Asia1型FMDV江苏/05谱系及新疆/03谱系毒株的二联PCR引物,优化单项和二联RT-PCR反应条件,并进行特异性及相关病毒检测。结果优化的单项和二联RT-PCR特异性均良好,检测9份FMDV,病料的结果与其基因序列测定结果完全一致。结论已建立了FMDVAsia1型江苏/05谱系和新疆/03谱系毒株的二联RT-PCR检测方法,为FMDV的诊断、流行病学调查及疫苗应用奠定了基础。Objective To develop a multiplex RT-PCR method for determination of foot-and-mouth disease virus （FMDV） type Asia 1 of XJ / 03 and JS / 05 topotypes. Methods The sequences of a large quantity of epidemic FMDV strains reported in GenBank were compared, based on which the primers for differential diagnosis of infection with FMDV type Asia 1 of XJ / 03 and JS / 05 topotypes by multiplex RT-PCR were designed, and the conditions for single and multiplex RT-PCR were optimized. The developed method was verified for specificity and used for determination of relevant viruses. Results Both the optimized single and multiplex RT-PCR methods showed high speeificities. The determination results of 9 FMDV samples by the developed method were completely consistent with those by gene sequencing. Conclusion A multiplex RT-PCR method for determination of FMDV type Asia 1 of XJ/ 03 and JS/05 topotypes was developed, which laid a foundation of diagnosis and epidemiological investigation of FMDV as well as application of FMD vaccine.

关 键 词：口蹄疫病毒 Asia1型 江苏/05谱系 新疆/03谱系 RT-PCR 

分 类 号：R379.4[医药卫生—病原生物学] R735.7[医药卫生—基础医学]