应用微卫星DNA对常染色体显性遗传性多囊肾病（ADPKD）进行症状前检测及产前诊断的研究
发布时间：2018-10-22 09:35
【摘要】：目的：常染色体显性遗传性多囊肾病(ADPKD)是一种常见的遗传性肾病,具有延迟显性遗传的特点。至今尚无有效的治疗方法,及时对ADPKD家系中的成员进行症状前基因检测和高风险胎儿进行产前诊断,是控制其发生和发展的关键。本实验应用与PKD1紧密连锁的微卫星DNA，通过家系连锁分析对一ADPKD家系进行基因定位和突变基因的筛查,完成常染色体显性多囊肾病的症状前检测及产前诊断,为临床分子诊断奠定基础。 方法：采用传统苯酚氯仿法提取ADPKD家系成员外周血的基因组DNA和胎儿绒毛膜绒毛DNA，选取4个与广西人PKD1紧密连锁的微卫星DNA（SM6、KG8、CW4、CW2）为遗传标志，荧光PCR扩增4个微卫星DNA，然后将荧光标记的PCR扩增产物进行毛细管电泳检测-基因扫描，通过基因连锁分析和单体型分析对一ADPKD家系共27人（包括1名胎儿）进行致病基因检测。 结果：27名家系成员中，Ⅰ1、Ⅱ1、Ⅱ3、Ⅱ8、Ⅱ10、Ⅱ12、Ⅲ3、Ⅲ9均为诊断明确的ADPKD患者，带有相同的与致病基因PKD1连锁的单倍型2—1—2；1名18岁个体Ⅲ5为PKD1基因突变症状前个体,携带有致病的2—1—2，无临床症状，处于囊肿发生前期；Ⅳ1不携带有2—1—2，产前诊断结果显示胎儿未获得致病基因，为正常后代，不会遗传该病，可继续妊娠。 结论：微卫星DNA具有高度多态性，，在同一家系内具有高度的遗传保守性，其基因片段短、扩增效率高、判型准确等特点使得该技术在ADPKD的疾病诊断简便、快速、灵敏。利用微卫星DNA进行多态性连锁分析,可在同一家系中筛选出ADPKD患者，它是目前临床上应用最多的一种ADPKD基因诊断方法。选取多个与致病基因连锁的微卫星DNA作为标记，对ADPKD家系成员进行症状前基因检测和产前诊断可提高确诊率。
[Abstract]:Objective: autosomal dominant polycystic kidney disease (ADPKD) is a common hereditary nephropathy with delayed dominant inheritance. Up to now, there is no effective treatment method. It is the key to control the occurrence and development of ADPKD pedigree to detect presymptomatic gene and prenatal diagnosis of high-risk fetus. In this study, microsatellite DNA, closely linked to PKD1, was used to detect and diagnose autosomal dominant polycystic kidney disease (autosomal dominant polycystic kidney disease) in a ADPKD pedigree by pedigree linkage analysis. To lay the foundation for clinical molecular diagnosis. Methods: genomic DNA and fetal chorionic villi DNA, were extracted from peripheral blood of ADPKD family members by traditional phenol chloroform method. Four microsatellite DNA (SM6,KG8,CW4,CW2) closely linked to Guangxi PKD1 were selected as genetic markers. Four microsatellite DNA, were amplified by fluorescence PCR, and then the fluorescent labeled PCR amplification products were detected by capillary electrophoresis-gene scanning. Genetic linkage analysis and haplotype analysis were used to detect pathogenic genes in a ADPKD family of 27 individuals (including 1 fetus). Results: among the 27 family members, 鈪
本文编号：2286790
[Abstract]:Objective: autosomal dominant polycystic kidney disease (ADPKD) is a common hereditary nephropathy with delayed dominant inheritance. Up to now, there is no effective treatment method. It is the key to control the occurrence and development of ADPKD pedigree to detect presymptomatic gene and prenatal diagnosis of high-risk fetus. In this study, microsatellite DNA, closely linked to PKD1, was used to detect and diagnose autosomal dominant polycystic kidney disease (autosomal dominant polycystic kidney disease) in a ADPKD pedigree by pedigree linkage analysis. To lay the foundation for clinical molecular diagnosis. Methods: genomic DNA and fetal chorionic villi DNA, were extracted from peripheral blood of ADPKD family members by traditional phenol chloroform method. Four microsatellite DNA (SM6,KG8,CW4,CW2) closely linked to Guangxi PKD1 were selected as genetic markers. Four microsatellite DNA, were amplified by fluorescence PCR, and then the fluorescent labeled PCR amplification products were detected by capillary electrophoresis-gene scanning. Genetic linkage analysis and haplotype analysis were used to detect pathogenic genes in a ADPKD family of 27 individuals (including 1 fetus). Results: among the 27 family members, 鈪
本文编号：2286790
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