【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。更多还原

【Abstract】 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 specificities. 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.更多还原