二重二次PCR对外显子缺失型DMD患者单淋巴细胞遗传学诊断的研究

Cytogenetic diagnosis on single lymphocyte of DMD patient with exon 50 deletion by two-time duplex PCR

目的对已知dystrophin基因50号外显子缺失的Duchenne型肌营养不良(Duchennemusculardystrophy,DMD)患者及正常对照者进行单个淋巴细胞遗传学诊断,建立该病单细胞植入前遗传学诊断(preimplantationgeneticdiag-nosis,PGD)方法,为携带者夫妇进行植入前遗传学诊断以提供细胞学基础。方法在解剖显微镜下获取已经确诊为dys-trophin基因50号外显子缺失型DMD患者和正常对照者的单个淋巴细胞,采用dystrophin基因50号外显子引物/SRY基因引物二重二次PCR的方法进行遗传学诊断。结果男性患者单个淋巴细胞中SRY基因阳性同时50号外显子阴性的比例为92%;正常男性对照中SRY基因和50号外显子均阳性的比例为91%;正常女性对照中50号外显子阳性和SRY基因阴性的比例为93%。结论通过二重二次PCR能够进行dystrophin基因50号外显子缺失型DMD患者和正常对照的单个淋巴细胞遗传学诊断,为PGD技术在该病遗传学诊断中的应用打下基础。

Objective To perform the cytogenetic diagnosis on single lymphocyte of DMD patient with dystrophin gene exon 50 deletion. Methods Single lymphoeytes of a DMD patient with dystrophin gene exon 50 deletion and normal volunteers were picked out and prepared for two-time duplex PCR. Results The rate of precise positive was 92% , 91% and 93% in specimens of the patient （SRY positive, exon 50 negative） , the male volunteer （SRY positive, exon 50 positive） and the female volunteer （SRY negative, exon 50 positive）, respectively. Conclusion Two-time duplex PCR is fit for the genetic diagnosis of single lymphocyte from DMD patient with dystrophin gene exon 50 deletion.