【摘要】 【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。更多还原

【Abstract】 To apply fluorescent PCR to detect the mutation of β Thalassemia genes and evaluate its effectiveness in preimplantation genetic diagnosis compared with the nested PCR. Fluorescent PCR and nested PCR followed by reverse dot blot (RDB) were applied to detect the β Thalassemia 41 42 mutation heterozygote from a single lymphocyte. The fluorescent PCR achieved amplification efficiency of 92.5%and allele drop out (ADO) rate of 8.8%. The nested PCR achieved amplification efficiency of 91.7%and ADO rate of 15.8%. There was no significant difference in the amplification efficiency between two protocols, but the ADO rate of fluorescent PCR was significantly lower than that of the nested PCR (P< 0.05). [Conclusion]Single cell fluorescent PCR can markedly decrease ADO rate and may be a powerful technique for clinical use of PGD for β Thalassemia.更多还原