何展文,陈启慧,李平甘,吴若豪,李栋方,李宇,周小琳,罗向阳

• 1:中山大学孙逸仙纪念医院

摘要(Abstract)：

目的探讨联合应用MLPA和二代基因测序技术对Duchenne型肌营养不良症分子遗传学诊断的作用。方法收集2012-01—2019-12在中山大学孙逸仙纪念医院儿童神经内分泌专科就诊经临床和肌肉活检明确诊断Duchenne型肌营养不良症患儿,联合应用MLPA和二代测序技术,分析其DMD基因单个外显子的缺失情况。结果纳入Duchenne型肌营养不良症患儿59例,均为男性,就诊年龄(4.72±2.42)岁。58例患儿确定了致病基因突变位点,基因诊断阳性率为98.30%。其中缺失突变28例(47.46%),以第45-55号外显子缺失频率最高;重复突变7例(11.86%),23例(38.98%)微小突变,其中包括无义突变8例(13.55%),移码突变6例(10.17%),错义突变6例(10.17%),微缺失3例(5.08%)。结论联合应用MLPA和二代测序技术是明确Duchenne型肌营养不良症致病基因突变位点最佳策略,能为Duchenne型肌营养不良症的精准诊治和家系成员的遗传咨询提供准确的依据。

关键词(KeyWords)： 肌营养不良;DMD基因;外显子;MLPA;二代测序;基因治疗

Abstract:

Keywords:

基金项目(Foundation): 2018年中山大学本科教学改革研究项目(编号:81000-18832601)

作者(Author): 何展文,陈启慧,李平甘,吴若豪,李栋方,李宇,周小琳,罗向阳

参考文献(References)：

• [1] MOAT S J,BRADLEY D M,SALMON R,et al.Newborn bloodspot screening for Duchenne muscular dystrophy:21 years experience in Wales (UK)[J].Eur J HumGenet,2013,21(10):1049-1053.DOI:10.1038/ejhg.2012.301.
• [2] WEIN N,ALFANO L,FLANIGAN K M.Genetics and emerging treatments for Duchenne and Becker muscular dystrophy[J].Pediatr Clin North Am,2015,62(3):723-742.DOI:10.2147/TACG.S8762.
• [3] ZIMOWSKI J G,PAWELEC M,PURZYCKA J K,et al.Deletions,not duplications or small mutations,are the predominante new mutations in the dystrophin gene[J].J Hum Genet,2017,62(10):885-888.DOI:10.1038/jhg.2017.70.
• [4] WANG Y,YANG Y,LIU J,et al.Whole dystrophin gene analysis by next-generation sequencing:a comprehensive genetic diagnosis of Duchenne and Becker muscular dystrophy[J].Mol Genet Genomics,2014,289(5):1013-1021.DOI:10.1007/s00438-014-0847-z.
• [5] AARTSMA-RUS A,GINJAAR I B,BUSHBY K.The importance of genetic diagnosis for Duchenne muscular dystrophy[J].J Med Genet,2016,53(3):145-151.DOI:10.1136/jmedgenet-2015-103387.
• [6] BIRNKRANT D J,BUSHBY K,BANN C M,et al.Diagnosis and management of Duchenne muscular dystrophy,part 1:diagnosis,and neuromuscular,rehabilitation,endocrine,and gastrointestinal and nutritional management[J].Lancet Neurol,2018,17:251-267.DOI:10.1016/S1474-4422(18)30024-3.
• [7] MOHAMMED F,ELSHAFEY A,AL-BALOOL H,et al.Mutation spectrum analysis of Duchenne/Becker muscular dystrophy in 68 families in Kuwait:the era of personalized medicine[J].PLoS One,2018,13(5):e0197205.DOI:10.1371/journal.pone.0197205.
• [8] LEE B L,NAM S H,LEE J H,et al.Genetic analysis of dystrophin gene for affected male and female carriers with Duchenne/Becker muscular dystrophy in Korea[J].J Korean Med Sci,2012,27(3):274-280.DOI:10.3346/jkms.2012.27.3.274.
• [9] TAKESHIMA Y,YAGI M,OKIZUKA Y,et al.Mutationspectrum of the dystrophin gene in 442 Duchenne/Becker muscular dystrophy cases from one Japanese referral center[J].J Hum Genet,2010,55(6):379-388.DOI:10.1038/jhg.2010.49.
• [10] KOLE R,KRIEG A M.Exon skipping therapy for Duchenne muscular dystrophy[J].Adv Drug Deliv Rev,2015,87:104-107.DOI:10.1016/j.addr.2015.05.008.
• [11] MCDONALD C M,WONG B,FLANIGAN K M,et al.Placebo-controlled Phase 2 Trial of Drisapersen for Duchenne Muscular Dystrophy[J].Ann Clin Transl Neurol,2018,5(8):913-926.DOI:10.1002/acn3.579.
• [12] GOEMANS N,MERCURI E,BELOUSOVA E,et al.A randomized placebo-controlled phase 3 trial of an antisense oligonucleotide,drisapersen,in Duchenne muscular dystrophy[J].Neuromuscul Disord,2018,28(1):4-15.DOI:10.1016/j.nmd.2017.10.004.
• [13] LIM K R,MARUYAMA R,YOKOTA T,et al.Eteplirsen in the treatment of Duchenne muscular dystrophy[J].Drug Des Devel Ther,2017,11:533-545.DOI:10.2147/DDDT.S97635.
• [14] CHARLESTON J S,SCHNELL F J,DWORZAK J,et al.Eteplirsen treatment for Duchenne muscular dystrophy:Exon skipping and dystrophin production[J].Neurology,2018,90(24):e2146-e2154.DOI:10.1212/WNL.0000000000005979.
• [15] WELCH E M,BARTON E R,ZHUO J,et al.PTC124 targets genetic disorders caused by nonsense mutations[J].Nature,2007,447(7140):87-91.DOI:10.1038/nature05756.
• [16] MCDONALD C M,CAMPBELL C,TORRICELLI R E,et al.Ataluren in patients with nonsense mutation Duchenne muscular dystrophy (ACT DMD):a multicentre,randomised,double-blind,placebo-controlled,phase 3 trial[J].Lancet,2017,390(10101):1489-1498.DOI:10.1016/S0140-6736(17)31611-2.
• [17] MERCURI E,MUNTONI F,OSORIO A N,et al.Safety and effectiveness of ataluren:comparison of results from the STRIDE Registry and CINRG DMD Natural History Study[J].J Comp Eff Res,2020,9(5):341-360.DOI:10.2217/cer-2019-0171.