常染色体显性多囊肾疾病行胚胎植入前遗传学诊断的实验研究  

Experimental research on preimplantation genetic diagnosis for autosomal dominant polycystic kidney disease

作　　者：朱琴[1] 徐炳森[1] 黄学锋[1] 周颖[1] 

机构地区：[1]温州医学院附属第一医院生殖医学中心,浙江温州325000

出　　处：《温州医学院学报》2009年第4期328-332,共5页Journal of Wenzhou Medical College

基　　金：温州医学院发展基金重大项目(2003001)

摘　　要：目的:建立由PKD1突变所致常染色体显性多囊肾疾病(autosomal dominant polycystic kidneydisease,ADPKD)的胚胎植入前遗传学诊断(preimplantation genetic diagnosis,PGD)方法。方法:①通过微卫星连锁分析确定2个多囊肾家系的ADPKD致病基因。检测的微卫星包括为与PKD1连锁的KG8、SM6、CW4和CW2以及与PKD2连锁的D4S1534、D4S1563、D4S414和D4S423。②对18个淋巴细胞和1个PKD1突变所致ADPKD成员行常规体外受精胚胎移植后的5个废弃胚胎15个卵裂球行多重巢式PCR和毛细管电泳检测与PKD1连锁的微卫星分型。结果:①KG8、CW4和CW2可作为连锁微卫星分析外周血和单个细胞的PKD1突变;②2个家系的致病基因均为PKD1;③单个卵裂球扩增成功率为86.67%(13/15),单个淋巴细胞扩增成功率为88.89%(16/18),CW4等位基因脱扣率为25%(4/16),CW2未发现等位基因脱扣,均未发现污染,2个胚胎携带致病基因。结论:PKD1连锁的微卫星分型可作为PKD1突变所致ADPKD的PGD诊断方法。Objective： To establish a method of preimplantation genetic diagnosis （PGD） for autosomal dominant polycystic kidney disease （ADPKD） caused by PKD1 mutation. Method： PCR and capillary electrophoresis were used to analysis microsatellite markers KGS, SM6, CW4 and CW2 linked to PKD1 and D4S1534, D4S1563, D4S414 and D4S423 linked to PKD2 in two ADPKD families to find out the possible mutation of the 2 families. Then, microsatellite typing was performed using multiplex nested PCR and capillary electrophoresis in 5 embryos （15 blastomeres）, which were discarded after in vitro fertilization and embryo transfer in an ADPKI） male caused by PKD1 mutation, and 18 lymphocytes from a normal donor. Result： Microsatellite markers KG8, CW4 and CW2 can be used for linkage analysis in peripheral blood and single cells. The linkage analyses confirmed that ADPKI） in the two families were caused by PKD1 mutation. The PCR amplification success rates of single blastomeres and single lymphocytes were 88.89% （13/15） and 86.67% （16/18）, respectively. The allele drop-out （ADO） rate of CW4 was 25% （4/16）, while ADO did not occurred in CW2. Contamination rate was zero. Two embryos were mutation-affected. Conclusion： The linkage analysis using microsatellite markers can be used in PGD of ADPKD caused by PKD1 mutation.

关 键 词：常染色体多囊肾疾病 PKD1 微卫星 植入前诊断 遗传学 

分 类 号：R715[医药卫生—妇产科学]