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CLC number: R394

On-line Access: 2019-08-05

Received: 2018-10-28

Revision Accepted: 2019-03-25

Crosschecked: 2019-07-09

Cited: 0

Clicked: 1755

Citations:  Bibtex RefMan EndNote GB/T7714

 ORCID:

Yan-mei Yang

https://orcid.org/0000-0001-6204-6600

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Journal of Zhejiang University SCIENCE B 2019 Vol.20 No.9 P.753-765

http://doi.org/10.1631/jzus.B1800541


Comprehensive genetic diagnosis of patients with Duchenne/Becker muscular dystrophy (DMD/BMD) and pathogenicity analysis of splice site variants in the DMD gene


Author(s):  Yan-mei Yang, Kai Yan, Bei Liu, Min Chen, Li-ya Wang, Ying-zhi Huang, Ye-qing Qian, Yi-xi Sun, Hong-ge Li, Min-yue Dong

Affiliation(s):  Department of Reproductive Genetics, Women’s Hospital, School of Medicine, Zhejiang University, Hangzhou 310006, China; more

Corresponding email(s):   dongmy@zju.edu.cn

Key Words:  Dystrophin gene, Variation, Genetic diagnosis, Splice site mutation, Hybrid minigene splicing assay


Yan-mei Yang, Kai Yan, Bei Liu, Min Chen, Li-ya Wang, Ying-zhi Huang, Ye-qing Qian, Yi-xi Sun, Hong-ge Li, Min-yue Dong. Comprehensive genetic diagnosis of patients with Duchenne/Becker muscular dystrophy (DMD/BMD) and pathogenicity analysis of splice site variants in the DMD gene[J]. Journal of Zhejiang University Science B, 2019, 20(9): 753-765.

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author="Yan-mei Yang, Kai Yan, Bei Liu, Min Chen, Li-ya Wang, Ying-zhi Huang, Ye-qing Qian, Yi-xi Sun, Hong-ge Li, Min-yue Dong",
journal="Journal of Zhejiang University Science B",
volume="20",
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pages="753-765",
year="2019",
publisher="Zhejiang University Press & Springer",
doi="10.1631/jzus.B1800541"
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%0 Journal Article
%T Comprehensive genetic diagnosis of patients with Duchenne/Becker muscular dystrophy (DMD/BMD) and pathogenicity analysis of splice site variants in the DMD gene
%A Yan-mei Yang
%A Kai Yan
%A Bei Liu
%A Min Chen
%A Li-ya Wang
%A Ying-zhi Huang
%A Ye-qing Qian
%A Yi-xi Sun
%A Hong-ge Li
%A Min-yue Dong
%J Journal of Zhejiang University SCIENCE B
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%I Zhejiang University Press & Springer
%DOI 10.1631/jzus.B1800541

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T1 - Comprehensive genetic diagnosis of patients with Duchenne/Becker muscular dystrophy (DMD/BMD) and pathogenicity analysis of splice site variants in the DMD gene
A1 - Yan-mei Yang
A1 - Kai Yan
A1 - Bei Liu
A1 - Min Chen
A1 - Li-ya Wang
A1 - Ying-zhi Huang
A1 - Ye-qing Qian
A1 - Yi-xi Sun
A1 - Hong-ge Li
A1 - Min-yue Dong
J0 - Journal of Zhejiang University Science B
VL - 20
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SP - 753
EP - 765
%@ 1673-1581
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PB - Zhejiang University Press & Springer
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DOI - 10.1631/jzus.B1800541


Abstract: 
Duchenne muscular dystrophy (DMD) and Becker muscular dystrophy (BMD) are caused by mutations in the DMD gene. The aim of this study is to identify pathogenic DMD variants in probands and reduce the risk of recurrence of the disease in affected families. variations in 100 unrelated DMD/BMD patients were detected by multiplex ligation-dependent probe amplification (MLPA) and next-generation sequencing (NGS). Pathogenic variants in DMD were successfully identified in all cases, and 11 of them were novel. The most common mutations were intragenic deletions (69%), with two hotspots located in the 5' end (exons 2–19) and the central of the DMD gene (exons 45–55), while point mutations were observed in 22% patients. Further, c.1149+1G>A and c.1150−2A>G were confirmed by hybrid minigene splicing assay (HMSA). This two splice site mutations would lead to two aberrant DMD isoforms which give rise to severely truncated protein. Therefore, the clinical use of MLPA, NGS, and HMSA is an effective strategy to identify variants. Importantly, eight embryos were terminated pregnancies according to prenatal diagnosis and a healthy boy was successfully delivered by preimplantation genetic diagnosis (PGD). Early and accurate genetic diagnosis is essential for prenatal diagnosis/PGD to reduce the risk of recurrence of DMD in affected families.

假肥大型肌营养不良症(DMD/BMD)遗传学诊断及剪接突变的致病性分析

目的:针对100例无亲缘关系的假肥大型肌营养不良症(DMD/BMD)患者,联合应用多种检测技术进行遗传学诊断,并且建立个体化产前诊断及胚胎植入前遗传学诊断方案,以最大限度地降低DMD/BMD患儿的出生.
创新点:通过minigene剪接实验分析DMD:c.1149+1G>A和c.1150−2A>G突变是否导致剪接异常,并确定剪接方式.
方法:收集100例无亲缘关系DMD/BMD患者的临床资料,应用多重连接依赖式探针扩增技术(MLPA)、第二代测序(NGS)、minigene剪接实验(HMSA)进行遗传学诊断,并通过单体型分析及性别鉴定进行胚胎植入前遗传学诊断.
结论:联合应用多种检测技术可以尽早地对患者进行遗传学诊断,为临床遗传咨询和产前诊断及胚胎植入前遗传学诊断提供了科学依据.

关键词: DMD基因;突变;遗传学诊断;剪接突变;Minigene剪接实验

Darkslateblue:Affiliate; Royal Blue:Author; Turquoise:Article

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[42]List of electronic supplementary materials

[43]Table S1 Oligonucleotide primers for detection of mutations in DMD gene

[44]Table S2 Deletions in DMD gene detected by MLPA

[45]Table S3 Duplications in DMD gene detected by MLPA

[46]Table S4 Point mutations in DMD gene detected by NGS

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