Best Practice Guidelines on molecular diagnostics in Duchenne/Becker muscular dystrophies

Abbs, Stephen; Tuffery‐Giraud, Sylvie; Bakker, Egbert; Ferlini, Alessandra; Sejersen, Thomas; Mueller, C.W.

doi:10.1016/j.nmd.2010.04.005

Cited by 91 publications

(89 citation statements)

References 24 publications

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“…Diagnostic procedures for DMD have made a great leap forward with the development of MLPA and capillary sequencing, in terms of mutation detection rate for both del/dup and small mutations [Abbs et al, 2010;Flanigan et al, 2003;Janssen et al, 2005;Lalic et al, 2005]. More recently, the development of omic approaches, such as CGH and targeted resequencing, have heralded the arrival of rapid extensive screening for all types of mutation in this enormous gene [Bonnal et al, 2010;del Gaudio et al, 2008;Hegde et al, 2008].…”

Section: Discussionmentioning

confidence: 99%

“…Both in-frame and out-of-frame mutations in the DMD gene (MIM# 300377) belong to a very heterogeneous mutation spectrum; in 75-80% of all cases this involves deletions/duplications (pathogenic copy number variations or CNVs), whereas the remaining 20-25% feature small mutations. About 2% of dystrophin mutations are atypical, and are generally due to deep intronic mutations that affect the splicing choices of the gene [Abbs et al, 2010].…”

Section: Introductionmentioning

confidence: 99%

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Rapid, comprehensive analysis of the dystrophin transcript by a custom micro-fluidic exome array

et al. 2012

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Duchenne and Becker muscular dystrophies are caused by mutations in the dystrophin gene. Both the enormous size of this gene and heterogeneous set of causative mutations behind these pathologies may hamper and even prevent accurate molecular diagnosis. Often RNA analysis is required not only to identify mutations escaping MLPA/CGH or exon sequencing but also to validate the functional effect of novel variations that may affect the exon composition of the DMD gene. We present the design and experimental validation of a new, simple, and easy-to-use platform we call FluiDMD. This platform is based on the Applied Biosystems 7900HT TaqMan(®) low-density array technology and is able to define the full-exon composition, profile the dystrophin isoforms present, establish changes in mRNA decay, and potentially identify all deletions/duplications and splicing affecting mutations contemporaneously. Moreover, we demonstrate that this system accurately detects the pathogenic effect of all dystrophin mutations belonging to any category, thereby highlighting the functional validation capacity of this system. The high efficacy and sensitivity of this tool in detecting mutations in the dystrophin transcript can be exploited in a variety of cells/tissues, in particular skin, which is harvested by causing minimum patient discomfort. We therefore propose FluiDMD as a validated diagnostic biomarker for molecular profiling of dystrophinopathies.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Rapid, comprehensive analysis of the dystrophin transcript by a custom micro-fluidic exome array

et al. 2012

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“…[9] MLPA is currently the technique of choice as a first-line screen to detect deletions/ duplications in routine DMD/BMD genetic diagnostics. [10] There are few published reports on DMD mutations in South African populations. Previous studies in this country have identified deletions in 50% (30/60) of affected boys, using the Southern blotting technique, [11] 42% (46/110) of affected boys using Southern blotting together with mPCR, [12] and 57% (39/68) of affected boys using the Chamberlain and Beggs mPCR approach.…”

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confidence: 99%

Genetic testing for Duchenne/Becker muscular dystrophy in Johannesburg, South Africa

et al. 2013

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Background. Genetic testing for Duchenne/Becker muscular dystrophy (DMD/BMD) mutations initially involved multiplex polymerase chain reaction (mPCR), which targeted two mutation hotspots in the gene and detected deletions in affected males. A newer technology, multiplex ligation-dependent probe amplification (MLPA), was introduced for diagnostic testing in 2007. Objectives. To evaluate MLPA relative to mPCR as a technique for DMD/BMD diagnostic testing and to establish whether the mutation profile in affected individuals differs between different South African ethnic groups. Methods. From January 2000 -May 2007, genetic diagnostic testing for DMD/BMD was undertaken in 128 male patients using mPCR. From May 2007 onwards, MLPA replaced this technique and 261 males were investigated. MLPA is a kit-based technology available from MRC-Holland. Results. Of the 128 and 261 probands tested using mPCR and MLPA, respectively, 31% and 34% were found to carry a deletion mutation. Further, MLPA could detect duplication mutations (11.5%), complex rearrangements (1.5%) and small mutations (1.5%). In black patients, deletion mutations were found to cluster in the 3' region of the gene. No population-specific pathogenic mutations were found. Conclusions. The mutation detection rate for mPCR and MLPA is similar for deletion mutations, but MLPA proved to be a better diagnostic approach as it could detect other types of mutations as well, including duplications, complex rearrangements and small mutations. MLPA could also diagnose mutation status in at-risk female relatives, which is not possible with mPCR.

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“…Mutations in dystrophin protein disrupt the open reading frame and prevent the full translation of its protein product, in ~65% of cases [30,36].…”

Section: Mutation Analysis Of Dmdmentioning

confidence: 99%

Application of Multiplex PCR for Detection of Duchunne Muscular Dystrophy: A Childhood Neuromuscular Disorder

Srivastava¹,

Srivastava²

2018

J Neurol Neurosci

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Duchenne muscular dystrophy (DMD) is an X-linked recessive disorder that affects 1 in 3,600 -6,000 males and is caused by mutation in the dystrophin gene. This is neuromuscular disorder with progressive muscle weakness, which predominantly affect males. Till date no absolute cure of the disease is available in clinical practice. Early diagnosis and timely management are requisite for DMD and it enhances the quality of life of patients. Various diagnostic approaches are available but due to accuracy, early detection and non-invasive method molecular tools are most remarkable in recent era. Multiplex PCR has emerged as one of the most convenient tools for screening of DMD in terms of its sensitivity, specificity, accuracy, cost effectiveness and time consumption. The current study emphasizes advantages and shortcomings of multiplex PCR with reference to most of the past studies along with its challenges for DMD detection in detail. Mutation detection is evidently crucial for diagnosis, as well as it may also be significant for future therapeutic purposes. Further research is important to elucidate specific mutation pattern in association with management and therapies of proband.

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Best Practice Guidelines on molecular diagnostics in Duchenne/Becker muscular dystrophies

Cited by 91 publications

References 24 publications

Rapid, comprehensive analysis of the dystrophin transcript by a custom micro-fluidic exome array

Rapid, comprehensive analysis of the dystrophin transcript by a custom micro-fluidic exome array

Genetic testing for Duchenne/Becker muscular dystrophy in Johannesburg, South Africa

Application of Multiplex PCR for Detection of Duchunne Muscular Dystrophy: A Childhood Neuromuscular Disorder

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