外周血基因表达分析内参基因的优选

摘要

目的 明确外周血RNA定量分析最佳内参基因组合.方法 采用实时荧光定量PCR方法检测5种管家基因的表达情况,应用qBASE+软件对基因表达稳定性进行分析并筛选最佳内参基因组合.结果 采用冠心病病例对照人群共98例,常用内参基因ACTB的表达水平变化最大.内参基因的表达稳定度由高到低依次为DECR1> GAPDH > TRAP1> PPIB >ACTB,并分析出最佳内参基因组合为DECR1、GAPDH和TRAP1.结论 研究结果为冠心病外周血的内参基因的选择提供了可靠的依据,可为该类样品的基因表达定量分析在基础研究和临床诊断的应用提供技术支撑.%Objective To identify the optimal set of the most stably expressed gene in human peripheral blood.Methods Quantitative real-time PCR was used to determine the transcription profiles of 5 housekeeping genes.qBASE+software was used to ana-lyze the gene's stabilities and choose the optimal set of reference genes.Results The total 98 case and control samples are collected from patients with coronary heart disease.The Ct range of ACTB,as the most commonly used reference gene,was changed widely from 20.09(30.06.The stability of these genes ranked in the following order:DECR1> GAPDH > TRAP1> PPIB >ACTB,and the optimal reference genes set was composed of DECR1,GAPDH,and TRAP1.Conclusion The results supply a reliable evidence for choosing the reference genes in human peripheral blood,and provide a technical support for application of gene expression analysis in the basic research and clinic diagnosis.