甲型H1N1流感病毒两种方法检测限的比较

The detection limit research of H1N1 influenza virus by RT-PCR and real-time pcr

目的:比较RT-PCR和Real time PCR检测甲型H1N1流感病毒的灵敏度。方法:体外转录生成甲型H1N1流感病毒M基因235 bp的保守序列作为阳性毒株RNA为模板,计算出相应的拷贝数后进行10倍梯度稀释,在引物探针的作用下分别进行RT-PCR和Real time PCR的检测。结果:Real time PCR的最低检测限为8 copies/μl,RT-PCR的最低检测限为80 copies/μl,线性关系好。结论:检测甲型H1N1流感病毒Real time PCR方法比RT-PCR灵敏度高10倍。

Objective：To compare the sensitivity for detection H1N1 influenza virus by RT-pcr and real-time pcr.Methods： Conserved Sequence of 235bp was transcribed in H1N1 influenza virus as positive control template,the concentration of which was ordinal diluted by 1：10 after the total numbers of copies was obtained.The two methods above were applied to continue detection with the primers and probes.Results： The detection limit of RT-pcr and real-time pcr was 80 copies/μl and 8 copies/μl respectively.Conclution： The sensitivity of real-time pcr method is 10 times higher than RT-pcr method.