Real-time quantitative RT-PCR for the detection of HLA-DRα mRNA expression

Objective To establish a real-time quantitative RT-PCR method for detection of HLA-DRα mRNA expression.Methods The vector containing HLA-DRα gene as standard template was constructed by T-A cloning technique.A real-time RT-PCR was established by used the fluorogenic probe(Taqman probe).HLA-DRα mRNA expression level in monocytes induced by achyrathes bidentata polysacchari-(des(ABPS)) was detected with real-time quantitative RT-PCR and semi-quantitative RT-PCR.Results The expression of HLA-DRα mRNA in monocytes induced by ABPS was increased significantly,but no significant change of HLA-DRα mRNA expression was fund in control group;The semi-quantitative RTPCR results also demonstrated the variety of HLA-DRα level as what the real-time fluorogenic quantitative RT-PCR detected,but less sensitive and accurate.Conclusion The fluorogenic real-time quantitative RT-PCR is more accurate and sensitive than semi-quantitive RT-PCR in Detection of HLA-DRα mRNA expression.