【摘要】 应用显微操作技术获取孕妇外周血中的单个有核红细胞 ,改良的PEP方法扩增单个有核红细胞的全基因组DNA ;在此基础上 ,应用荧光标记聚合酶链反应扩增 9个微卫星片段 ,进行基因型分析判定单个有核红细胞来源。综合性别和DMD基因内的数个STR位点连锁分析进行DMD基因诊断 ,应用PCR -STR连锁分析进行PKU基因诊断。结果显示 ,对 10例DMD高危胎儿中的 6例成功地进行了无创性产前基因诊断。同时对 1例PKU也成功地进行了无创性产前基因诊断。改良的PEP方法扩增单个细胞的全基因组可以满足基因诊断的要求 ,是无创性产前基因诊断中一种极有价值的全基因组扩增的方法更多还原

【Abstract】 We investigated the feasibility of using improved primer extension preamplification method to diagnose DMD and PKU. The fetal nucleated red blood cells from the peripheral blood of pregnant women were detected and individually retrieved into glass capillary pipettes using a micromanipulator under microscopic observation. The whole genome of a single cell was amplified by improved primer extension preamplification (PEP).Genotypes were analyzed by amplifying the 9 STR fragments using fluorescence PCR technique and NRBC’s(nucleated red blood cell) origin was determined.We diagnosed DMD prenatally using sex determination and linkage analysis of several STR sites of dystrophin,and we diagnosed PKU prenatally using PCR STR linkage analysis.6 of 10 potential DMD patients were diagnosed,including 1 male fetal patient,1 potential PKU patient was also diagnosed.The improved PEP method is a very valuable method of amplifying the whole genome of single cells,and the products of amplification are enough to the requirements of DNA in non invasive prenatal diagnosis.更多还原