應(yīng)用微衛(wèi)星DNA對常染色體顯性遺傳性多囊腎�。ˋDPKD）進(jìn)行癥狀前檢測及產(chǎn)前診斷的研究
發(fā)布時間：2018-10-22 09:35
【摘要】：目的：常染色體顯性遺傳性多囊腎病(ADPKD)是一種常見的遺傳性腎病,具有延遲顯性遺傳的特點(diǎn)。至今尚無有效的治療方法,及時對ADPKD家系中的成員進(jìn)行癥狀前基因檢測和高風(fēng)險(xiǎn)胎兒進(jìn)行產(chǎn)前診斷,是控制其發(fā)生和發(fā)展的關(guān)鍵。本實(shí)驗(yàn)應(yīng)用與PKD1緊密連鎖的微衛(wèi)星DNA，通過家系連鎖分析對一ADPKD家系進(jìn)行基因定位和突變基因的篩查,完成常染色體顯性多囊腎病的癥狀前檢測及產(chǎn)前診斷,為臨床分子診斷奠定基礎(chǔ)。 方法：采用傳統(tǒng)苯酚氯仿法提取ADPKD家系成員外周血的基因組DNA和胎兒絨毛膜絨毛DNA，選取4個與廣西人PKD1緊密連鎖的微衛(wèi)星DNA（SM6、KG8、CW4、CW2）為遺傳標(biāo)志，熒光PCR擴(kuò)增4個微衛(wèi)星DNA，然后將熒光標(biāo)記的PCR擴(kuò)增產(chǎn)物進(jìn)行毛細(xì)管電泳檢測-基因掃描，通過基因連鎖分析和單體型分析對一ADPKD家系共27人（包括1名胎兒）進(jìn)行致病基因檢測。 結(jié)果：27名家系成員中，Ⅰ1、Ⅱ1、Ⅱ3、Ⅱ8、Ⅱ10、Ⅱ12、Ⅲ3、Ⅲ9均為診斷明確的ADPKD患者，帶有相同的與致病基因PKD1連鎖的單倍型2—1—2；1名18歲個體Ⅲ5為PKD1基因突變癥狀前個體,攜帶有致病的2—1—2，無臨床癥狀，處于囊腫發(fā)生前期；Ⅳ1不攜帶有2—1—2，產(chǎn)前診斷結(jié)果顯示胎兒未獲得致病基因，為正常后代，不會遺傳該病，可繼續(xù)妊娠。 結(jié)論：微衛(wèi)星DNA具有高度多態(tài)性，，在同一家系內(nèi)具有高度的遺傳保守性，其基因片段短、擴(kuò)增效率高、判型準(zhǔn)確等特點(diǎn)使得該技術(shù)在ADPKD的疾病診斷簡便、快速、靈敏。利用微衛(wèi)星DNA進(jìn)行多態(tài)性連鎖分析,可在同一家系中篩選出ADPKD患者，它是目前臨床上應(yīng)用最多的一種ADPKD基因診斷方法。選取多個與致病基因連鎖的微衛(wèi)星DNA作為標(biāo)記，對ADPKD家系成員進(jìn)行癥狀前基因檢測和產(chǎn)前診斷可提高確診率。
[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
本文編號：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
本文編號：2286790
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