MLPA及测序技术在DMD/BMD家系分析中的应用  

Genealogy analysis of duchenne muscular dystrophy by multiplex ligation-dependent probe amplification and sequencing technology

作　　者：古艳[1] 谢建生[2] 都莉[1] 韩春锡[3] 万琼[1] 

机构地区：[1]江西省南昌大学第三附属医院检验科,江西南昌330006 [2]广东省深圳市妇幼保健院产前诊断中心,广东深圳518048 [3]广东省深圳市儿童医院神经内科,广东深圳518048

出　　处：《国际检验医学杂志》2012年第24期2976-2977,共2页International Journal of Laboratory Medicine

摘　　要：目的运用MLPA技术和DNA及cDNA测序技术对DMD/BMD进行家系分析,对患者、可能携带者基因诊断并探讨诊断流程的临床可行性。方法对2个DMD/BMD家系中6例患者、13例女性可能携带者、6例男性家系成员,6例女性和男性健康对照共31例采集外周血提取DNA,运用MLPA技术分析对以上31例的DMD基因79个外显子;患者取右侧腓肠肌10～30mg肌肉提取RNA,逆转录cDNA;分别进行DNA及cDNA序列测定,测序结果与MLPA结果进行比较。结果经MLPA检测,家系1的4例患者均缺失DMD基因Exon50,家系2中2例患者均缺失Exon43。以上结果经肌肉cDNA测序证实了相应外显子缺失。结论 MLPA技术结合DNA及cDNA测序技术进行DMD家系分析具有可靠的临床应用价值。Objective To analyze the DMD genealogy by MLPA technique in combine with DNA and cDNA sequencing technology.Methods There were 31 individuals accepted DMD gene diagnosis,including 6 DMD/BMD patients,13 possible carriers and 6 healthy men in 2 DMD/BMD families,moreover 6 healthy women and men were selected from health examination people.Genomic DNA of the peripheral blood was extracted from the pedigrees′ members with DMD.RNA was extracted from the bioptic muscle of the DMD patients and was reversed transcription to cDNA.Gene diagnosis was performed for theses pedigrees′ members using MLPA technique,the mutation was analyzed applying with DNA and/or cDNA sequence technique.Simultaneously,compare the effects of these methods on detecting DMD gene deletion.Results 4 patients of the first DMD family was deleted exon50,and the fetus was confirmed no DMD exons deletion.2 patients were found deletion exon43 in the second family.MLPA analysis、DNA and cDNA sequencing technology showed the same result.Conclusion MLPA in company with DNA and cDNA sequencing technology could applied into clinical gene diagnosis for DMD.

关 键 词：肌营养不良 核酸探针 基因扩增 系谱 

分 类 号：R685[医药卫生—骨科学]