应用荧光PCR技术检测单细胞β地中海贫血基因

• 作 者: 邓捷 庄广伦 彭文林 周灿权 梁晓燕 詹前胜 孙宏钰 陈勇 曾艳红
• 机 构: 中山大学附属第一医院生殖中心,中山大学附属第一医院生殖中心,中山大学附属第一医院生殖中心,中山大学附属第一医院生殖中心,中山大学附属第一医院生殖中心,中山大学附属第一医院生殖中心,中山大学法医学教研室
• 出 处: 中山大学学报(医学科学版)
• 发表日期: 2004年02期
• 关键词: β地中海贫血 聚合酶链反应(荧光) 植入前诊断

【目的】应用荧光PCR技术检测单细胞β地中海贫血(β地贫)基因,并与巢式PCR相比较,探讨荧光PCR应用于β地贫植入前诊断中的可行性。【方法】获取β地贫41-42突变杂合子单个淋巴细胞,用巢式PCR及荧光PCR技术检测β珠蛋白基因。【结果】160个淋巴细胞进行荧光PCR扩增,β地贫基因的平均扩增效率为92.5%,等位基因脱扣(ADO)率为8.8%;240个淋巴细胞进行巢式PCR扩增,β地贫基因的扩增效率为91.7%,ADO率为15.8%。两种方法的扩增效率没有统计学差异,但荧光PCR的ADO发生率显著低于巢式PCR的ADO发生率(P<0.05)。【结论】本研究建立的单细胞荧光PCR技术具有较高的扩增效率,可显著降低ADO发生率,可望用于β地贫的临床植入前诊断。

【Objective】 Fluorescent PCR was used to detect single-cell β-thalassemia (β-thalassemia) gene and compared with nested PCR to explore the feasibility of fluorescence PCR in the diagnosis of β-thalassemia. 【Method】 Single lymphocyte of heterozygote of 41-42 mutation in β-thalassemia was obtained. The β-globin gene was detected by nested PCR and fluorescent PCR. 【Results】 160 lymphocytes were amplified by fluorescence PCR. The average amplification efficiency of β thalassemia gene was 92.5% and the rate of allele transfer (ADO) was 8.8%. 240 lymphocytes were amplified by nested PCR, β thalassemia gene amplification efficiency was 91.7%, ADO rate was 15.8%. There was no statistical difference in amplification efficiency between the two methods, but the incidence of ADO in fluorescence PCR was significantly lower than that of nested PCR (P <0.05). 【Conclusion】 The single-cell fluorescence PCR technique established in this study has high amplification efficiency, can significantly reduce the incidence of ADO, and is expected to be used in clinical pre-implantation diagnosis of β-thalassemia.