联合基因测序与STR连锁分析进行苯丙酮尿症产前诊断的研究  

Study on prenatal diagnosis of PKU with gene sequencing combined with STR linkage analysis

作　　者：唐新华[1] 陈红[1] 苏洁[1] 章印红[1] 徐庆华[1] 朱宝生[1] 

机构地区：[1]昆明医科大学附属昆华医院,云南省第一人民医院遗传诊断中心,云南省出生缺陷与遗传病研究重点实验室

出　　处：《中国妇幼保健》2014年第6期927-930,共4页Maternal and Child Health Care of China

基　　金：昆明医科大学-云南省科技厅联合基金〔2010CD198〕

摘　　要：目的:建立稳定可靠的苯丙酮尿症产前基因诊断方法。方法:采集3个经典型PKU家系血样和胎儿羊水并提取gDNA,进行PAH基因13个外显子测序,通过测序技术直接诊断胎儿PAH基因突变,并同时使用4个与PAH基因紧密连锁的STR位点,分析PAH基因在家系中的传递。结果:家系1中胎儿与先证者不同,测序检出其未获得父亲携带并遗传给了先证者的c.728G>A突变,胎儿为突变携带着,后分娩一正常婴儿;家系2中先证者检出c.1068C>A和c.842+4A>T杂合突变,胎儿检出携带父源的c.1068C>A,未遗传到母源的c.842+4A>T,为携带者,连锁分析也证实胎儿遗传了母亲正常的PAH基因。该胎儿出生后正常。家系3中,PAH基因测序未能找到先证者PAH基因的2个突变,仅仅检出先证者和胎儿均携带遗传自母亲的c.690-691insG移码突变,但STR连锁分析因多态性差无法提供信息,导致无法得到确诊结论,加上孕妇就诊时已中孕期,没有时间尝试新的诊断技术寻找潜在的基因异常,后报告胎儿至少为携带者并遗传咨询,孕妇知情选择终止妊娠。结论:联合使用基因测序与STR位点连锁分析技术,是完成苯丙酮尿症产前基因诊断"双保险"技术体系,准确可靠,值得推广应用。Objective: To establish an stable and reliable method of PKU prenatal genetic diagnosis. Methods: Three different families with PKU history received PKU prenatal genetic diagnosis. Genomic DNA was extracted from the whole blood of all family numbers and amniotic fluid of fetuses,then exon 1- 13 of PAH gene were sequenced to find the mutations on PAH gene,at the same time,four STR sites linked with PAH gene were chosen to accomplish the PAH gene linkage analysis on those three families. Results: In family 1,a mutation occurred,c. 728G > A was found on proband but not on the fetus,which implied that the fetus would be protected by the normal PAH copy. In family 2,the fetus was detected with a mutation of c. 1068C > A on one PAH gene inherited from its father,and also a normal PAH gene from its mother,who carrying a mutation of c. 842 + 4A > T on one PAH gene. The fetus that mutation carrier was also proven by linkage analysis. In family 3,an uncertain report had to be delivered because that sequencing of PAH gene failed to find both two mutations on the PAH gene of proband,accompanying with a poor polymorphisms of PAH- STR sites,which could not provide a clear result for the fetus, but it was sure that the fetus was at least a carrier of a mutation of c. 690- 691insG on PAH gene. The pregnant women chose abortion after genetic consulting. Conclusion: Combining use of gene sequencing and PCR- STR is an accurate and reliable method in PKU prenatal diagnosis,which is worthy to be applied and popularized widely.

关 键 词：苯丙酮尿症 产前诊断 基因诊断 

分 类 号：R714.54[医药卫生—妇产科学]