论文部分内容阅读
目的 探讨杜氏肌营养不良症 (Duchenne muscular dystrophy,DMD)的无创性产前基因诊断的可行性。方法 用不连续密度梯度离心方法初步富集妊娠 9～ 2 1周孕妇外周血中的有核红细胞 ,细胞涂片离心机制片 ,瑞氏姬姆萨染色标记 ,显微操作仪获取单个有核红细胞 ,改良的 PEP(primer extensionpreamplification)方法扩增单个有核红细胞的全基因组 DNA;在综合性别和 DMD基因内的数个 STR位点连锁分析进行 DMD基因诊断的同时 ,鉴定单个有核红细胞的来源 ,再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 ,进一步判定单个有核红细胞来源。结果 成功诊断了 1例 DMD男性患病胎儿。结论 初步建立了 DMD的无创性产前基因诊断的方法。
Objective To investigate the feasibility of noninvasive prenatal diagnosis of Duchenne muscular dystrophy (DMD). Methods The nucleated erythrocytes in peripheral blood of 9-21 weeks pregnant women were pre-enriched by discontinuous density gradient centrifugation. The cells were smeared with Reichega Giemsa stain and labeled with a micromanipulator. Single nucleated erythrocytes , Improved PEP (primer extensionprepmplification) method to amplify the whole genome DNA of a single nucleated erythrocytes; in the analysis of the DMD gene with linkage analysis of several STR loci in the sex and DMD genes, the origin of single nucleated erythrocytes was identified, Nine microsatellite loci were amplified by fluorescence-labeled polymerase chain reaction (PCR-RFLP). Genotype analysis was performed to further determine the origin of single nucleated erythrocytes. Results One fetus with DMD was diagnosed successfully. Conclusion The method of noninvasive prenatal diagnosis of DMD was preliminarily established.