目的:建立SYBR GreenⅠ实时定量检测TCR Vγ亚家族T细胞表达水平的比较Ct法。方法:利用倍比稀释的成人正常胸腺组织cDNA建立TCR VγⅠ~VγⅢ和β2微球蛋白基因(β2M)基因的相对标准曲线,实验证实靶基因(VγⅠ~VγⅢ)分别和看家基因(β2M)的扩增效率一致,可采用比较Ct法对TCR VγⅠ~VγⅢ的表达进行相对定量,并运用该方法检测20例健康成人外周血TCR VγⅠ~VγⅢ的表达水平。结果:成人外周血TCR VγⅡ的表达量最高,TCR VγⅠ次之,TCRVγⅢ的表达量最低,三者比较有统计学意义(P<0.05)。结论:本研究率先建立了利用SYBR Green Ⅰ实时荧光定量RT-PCR检测人TCR Vγ基因mRNA表达水平比较Ct法,并在国内首先提供了健康成人外周血TCR Vγ基因各亚家族mRNA表达水平的资料。 Objective: To establish a real-time quantitative detection of TCR Vγ subfamily T cells by SYBR Green Ⅰ Ct method. Methods: The relative standard curve of TCR VγⅠ ~ VγⅢ and β2 microglobulin gene (β2M) gene was amplified by cDNA from adult normal thymus tissue. The target gene (VγⅠ ~ VγⅢ) was compared with that of housekeeping gene The efficiency of amplification was consistent. Relative expression of TCR VγⅠ-VγⅢ in TCR VγⅠ-VγⅢ could be quantified by Ct method. The expression of TCR VγⅠ-VγⅢ in peripheral blood of 20 healthy adults was detected by this method. Results: The expression of TCR VγⅡ in adult peripheral blood was the highest, followed by TCR VγⅠ, and TCRVγⅢ was the lowest, with statistical significance (P <0.05). Conclusion: This study established the first using SYBR Green Ⅰ real-time fluorescent quantitative RT-PCR detection of human TCR Vγ gene mRNA expression levels compared Ct method and the first to provide healthy adult peripheral blood TCR Vγ gene subfamily mRNA expression levels .