汉城病毒的TaqMan-BHO探针实时荧光定量检测方法的建立

Establishment of TaqMan-BHQ probe real-time fluorescence RT-PCR for quantitative detection of Soul virus

目的建立一种汉城病毒实时荧光定量RT-PCR快速的检测方法。方法用专业软件设计引物和TaqMan-BHQ探针,以人工合成L基因的片段作为模板,进行实时荧光定量RT-PCR研究。结果模板的Ct值与模板稀释浓度的对数存在良好的线性关系,标准曲线为Y=-3.607 X+41.84,r2=0.998,PCR扩增效率为108.1%,其最低检出限为53.2copies/μL。结论应用TaqMan-BHQ1探针的实时荧光RT-PCR检测汉城病毒核酸,具有耗时短、灵敏度高等特点。

Objective To establish a rapid method of real-time fluorescence RT-PCR to quantify Soul virus.Methods The pro-fessional software was adopted to design the primer and the TaqMan-BHQ probe.With artificially synthesized L gene segment as the template of Soul virus,the real-time RT-PCR for detecting Soul virus was researched.Results The Ct value of templates had a good linear relationship with the log value of the template diluted concentration.The standard curve was Y =-3.607X ＋41.84, r2 =0.998,the PCR amplification efficiency was 108.1%,its lowest detection limit was 53.2 copies/μL.Conclusion Applying the real-time fluorescence RT-PCR by the TaqMan-BHQ probe for detecting nucleic acid of Seoul virus has the characteristics of short time-consuming and high sensitivity.