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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 of RT-PCR and Real time PCR in detecting influenza A (H1N1) virus. Methods: The 235 bp sequence of M gene of influenza A (H1N1) virus was transcribed in vitro as a template of positive strain RNA. After corresponding copy number was calculated, a 10-fold gradient dilution was performed. The primers were used to perform RT-PCR And Real time PCR detection. Results: The detection limit of Real time PCR was 8 copies / μl, and the detection limit of RT-PCR was 80 copies / μl. The linearity was good. Conclusion: The Real-time PCR assay for detection of influenza A (H1N1) virus is 10-fold more sensitive than RT-PCR.