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ID:15215751
大小:43.50 KB
页数:14页
时间:2018-08-02
《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》由会员上传分享,免费在线阅读,更多相关内容在学术论文-天天文库。
1、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作者:朱琴,徐炳森,黄学锋,周颖【摘要】目的:建立由PKD1突变所致常染色体显性多囊肾疾病(autosomaldominantpolycystickidneydisease,ADPKD)的胚胎植入前遗传学诊断(preimplantationgeneticdiagnosis,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D4S42
2、3。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AD
3、PKD的PGD诊断方法。【关键词】常染色体多囊肾疾病;PKD1;微卫星;植入前诊断;遗传学14 Abstract:Objective:Toestablishamethodofpreimplantationgeneticdiagnosis(PGD)forautosomaldominantpolycystickidneydisease(ADPKD)causedbyPKD1mutation.Method:PCRandcapillaryelectrophoresiswereusedtoanalysismicrosatellitemarkersKG8
4、,SM6,CW4andCW2linkedtoPKD1andD4S1534,D4S1563,D4S414andD4S423linkedtoPKD2intwoADPKDfamiliestofindoutthepossiblemutationofthe2families.Then,microsatellitetypingwasperformedusingmultiplexnestedPCRandcapillaryelectrophoresisin5embryos(15blastomeres),whichwerediscardedafterinvit
5、rofertili-zationandembryotransferinanADPKDmalecausedbyPKD1mutation,and18lymphocytesfromanormaldonor.Result:MicrosatellitemarkersKG8,CW4andCW2canbeusedforlinkageanalysisinperipheralbloodandsinglecells.ThelinkageanalysesconfirmedthatADPKDinthetwofamilieswerecausedbyPKD1mutati
6、on.ThePCRamplificationsuccessratesofsingleblastomeresandsinglelymphocyteswere88.89%(13/15)and86.67%(16/18),respectively.Thealleledrop-out(ADO)rateofCW4was25%(4/16),whileADOdidnotoccurredinCW2.Contaminationratewaszero.Twoembryosweremutation-affected.Conclusion:Thelinkageanal
7、ysisusingmicrosatellitemarkerscanbeusedinPGDofADPKDcausedbyPKD1mutation.14 Keywords:ADPKD;PKD1;microsatellite;preimplantationdiagnosis;genetic 常染色体显性多囊肾疾病(autosomaldomi-nantpolycystickidneydisease,ADPKD)是最常见的常染色体显性遗传的单基因遗传病,人群发病率为1/1000[1]。ADPKD的主要表现为肾脏进行性囊性改变,常导致终末期肾病,居我
8、国终末期肾病病因的第4位[2]。ADPKD具有延迟性发病的特点,其症前检查结果很少影响未生育患者的生育意向,使其成为最易遗传给下一代的严重遗传病之一。因此,对ADP
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