Establishment of a TaqMan real-time quantitative reverse transcription-polymerase chain reaction assay for rubella virus RNA

Objective:To establish a real-time quantitative reverse transcription-polymerase chain reaction(RT-PCR) for rubella virus(RV),a rubella virus specific PCR amplicon as external standard to analytically estimate this real time quantitative assay.Methods: Firstly,the primer and TaqMan probe concentrations,as well as reaction temperatures were optimized to establish an efficient real-time quantitative RT-PCR assay for RV RNA.Next,and a RV specific PCR amplicon was made as external standard.Results: The RV specific PCR amplicon prepared for evaluation of the assay was 503bp,and its original concentration was 2.75×109 copies/μl.The real time quantitative assay was shown to have good linearity(r2=0.9986,P0.001).Conclusion: The established quantitative RT-PCR is a simple,rapid,quantitative and accurate assay for RV RNA.