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目的采用实时定量RT-PCR技术检测β地中海贫血(β地贫)珠蛋白基因的表达。方法选取β地中海贫血患者及正常成年人的样本DNA各100份,提取样本RNA,根据荧光实时定量RT-PCR引物及探针的设计要求,设计合成α、β、γ3对引物与3条荧光探针,建立荧光实时定量RT-PCR扩增体系,检测入选者α、β、γ珠蛋白基因mRNA的相对含量。结果对照组βmRNA/αmRNA明显大于轻型β地贫组与重型β地贫组,γmRNA/αmRNA、γmRNA/(βmRNA+γmRNA)明显小于其他2组,差异均有统计学意义(P<0.05)。结论β地中海贫血患者β珠蛋白基因表达水平较正常人低,γ珠蛋白基因表达水平较正常人高;随着β珠蛋白基因表达水平的降低,γ珠蛋白基因表达水平升高;不同类型β地中海贫血α、β、γ珠蛋白基因表达不同且存在竞争性反馈调节机制。
Objective To detect the expression of β-thalassemia globulin by real-time quantitative RT-PCR. Methods 100 samples of β-thalassemia patients and normal adults were collected and their RNAs were extracted. According to the design requirements of real-time quantitative RT-PCR primers and probes, the designed and synthesized primers of α, β, γ3 and three fluorescent probes The fluorescence quantitative real-time RT-PCR amplification system was established to detect the relative content of α, β, γ-globin gene mRNA. Results The mRNA level of βmRNA / αmRNA in the control group was significantly higher than those in the β - thalassemia group and the β thalassemia group. The levels of γmRNA / αmRNA and γmRNA / (γmRNA / γmRNA) in the control group were significantly lower than those in the other two groups (P <0.05). Conclusion β-thalassemia patients with β-globin gene expression levels lower than normal, γ-globin gene expression levels higher than normal; with the decline of β-globin gene expression, γ-globin gene expression levels; different types of β Thalassemia α, β, γ-globin gene expression different and there is a competitive feedback regulatory mechanism.