基于MLPA技术对单基因遗传病DMD/BMD、SMA基因诊断的研究

MLPA-Based gene diagnostic methods for single gene disorders DMD/BMD and SMA

目的建立单基因遗传病DMD/BMD与SMA的多重连接依赖性探针扩增技术(MLPA)检测平台,提高患者诊断率和携带者检出率,并指导再生育。方法选择2010年至2013年在解放军四六三医院就诊的50例DMD患者,5例BMD患者和7例SMA患者,运用MLPA技术对患者的DMD基因、SMN1基因的缺失/重复突变进行突变筛查,同时对有基因缺失的DMD、SMA先症者的姐妹或母亲进行DMD、SMN1基因携带者筛查。结果 DMD:55例患者中30例患者有外显子缺失,3例有外显子重复,其中20例外显子缺失的患者中8例其姐妹或母亲为缺失携带者。SMA:7例患者中5例SMN1第7+8号外显子纯合缺失和1例7外显子纯合缺失,1例没有缺失,5例患儿的父母亲中4例为SMN1外显子7+8部分缺失携带者。结论 MLPA是一种高效、灵敏、准确、性价比较高的DMD/BMD、SMA分子诊断新方法。MLPA的应用不依赖患者信息,可有效对DMD/BMD、SMA致病基因拷贝数进行相对定量,检出遗传缺陷携带者和提高患者诊断率。

Objective：To establish MLPA test platform for gene diagnosis of single gene disorders（DMD/BMD,SMA）,and to improve the detection rate of patients and carriers,and to direct reprod uction. Methods：From 2010 to 2013,50 cases of DMD,5 cases of BMD and 7cases of SMA patients in the 463 Hospital of PLA using MLPA technology detect DMD gene and SMN1 gene in patients with mutation screening,but also DMD and SMN1 gene carriers were screened for gene deletion in DMD,SMA sisters and mothers of the probands. Results：DMD：30 were found to have exon deletion mutations,3 were found to have exon duplication mutations of the dystrophin gene in 55 cases.Inaddition to 8 carriers were identified within 20 cases with deletion mutation by the method.SMA：MLPA analysis showed the same that 5 patients with exon 7 and 8 homozygous deletion,and 1patients with only exon 7 homozygous deletion of SMN1. The other 1cases presented no homozygous deletion,but 4 mothers of them were detected heterozygous by MLPA. Conclusion：MLPA is an efficient,sensitive,accurate and cost-effective method for DMD/BMD,SMA molecular diagnosis.Its application is independent from the information of patients.MLPA could relatively quantify the gene copy number efficiently,detect carrier with genetic defects and improve the diagnostic rate of patients.