杜氏肌营养不良症的无创性产前基因诊断研究

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9－21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，显微操作仪获得单个有核红细胞，改良的PEP（primer extension preamplification)方法扩增单个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诊断诊断的同时，鉴定单个有核红细胞的来源，再应用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。

Non-invasive prenatal diagnosis of Duchenne muscular dystrophy

OBJECTIVE: This paper was designed to investigate the feasibility of non-invasive prenatal diagnosis of Duchenne muscular dystrophy(DMD). METHODS: The nucleated red blood cells(NRBC) were separated with percoll using a discontinuous density gradient method. The cells were smeared on microscope slides using a cyto-centrifuge and then stained by Wright- Giemsa. NRBCs 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). The procedures for making prenatal diagnosis of DMD and determining the origin of NRBCs proceeded at the same time using sex determination and linkage analysis of several STR loci of dystrophin. Genotypes were analyzed by amplifying the 9 STR fragments using fluorescence-PCR technique and NRBCs origin was further determined. RESULTS: A case of DMD in male fetus was diagnosed. CONCLUSION: With the use of the method reported, the non-invasive prenatal diagnosis of DMD is possible.