Identification of deletions and duplications in the Duchenne muscular dystrophy gene and female carrier status in western India using combined methods of multiplex polymerase chain reaction and multiplex ligation-dependent probe amplification

Dastur, Rashna S; Kachwala, Munira Y; Khadilkar, Satish V; Hegde, Madhuri; Gaitonde, Pradnya S

doi:10.4103/0028-3886.91355

Cited by 11 publications

(8 citation statements)

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“…Out of 41 patients, 14 patients (34.1%) showed deletion, which is much lower than the proportion of deletions reported in different population of India using same methodology (multiplex PCR method). There is a wide variation in reported frequency of deletion in various parts of India and it ranges from 62.1% to 74% (8)(9)(10). Such wide variation in deletion rate is attributed to population based differences in mutations of dystrophin gene which could be due to accumulation of differences in intronic sequences and their distribution of differences over a period of time as a consequence of genetic drift.…”

Section: Discussionmentioning

confidence: 99%

Duchenne muscular dystrophy: A immunohistochemical profile and deletion pattern in dystrophin gene in North Indian population

et al. 2017

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Background: Duchenne muscular dystrophy (DMD), one of the most common X linked muscular disorder, affecting 1 in 3500 male births and is caused by mutation in dystrophin gene. 65% of DMD cases are caused by large deletion of dystrophin gene, followed by duplication (5-10%) or point mutation (25-30%). There is wide mutation spectrum of the mutations in dystrophin gene. Hence, population specific information is needed on mutation spectrum and frequency of common mutations occurring in that particular population for appropriate counseling, prenatal diagnosis and for developing genetic therapy in future. Aims and Objectives: To find out the frequency and distribution of deletion in dystrophin gene in DMD patients along with contribution of pathology and genetic testing in diagnosis of DMD and Becker muscular dystrophy (BMD) in North Indian population. Materials and Methods: Dystrophin gene was screened for deletion by multiplex polymerase chain reaction (PCR). Out of 41 patients, 09 patients underwent muscle biopsy, on which immunohistochemistry was performed for dystrophin, sarcoglycan, dysferlin and merosin. Results: Majority of the deletions were located in distal hotspot region (26/39 ~66.66%) which includes the exons 45-55 and 15.38% of deletions were located at the proximal hotspot region (2-19 exons). Conclusion: In the present study, 34% patients only showed deletion. Hence complete work up of any muscular dystrophy requires immnohistochemical analysis to see the expression of muscle proteins along with multipleplex PCR test to detect any exon deletion, multiplex ligation-dependent probe amplification (MLPA) to detect point mutation and duplication and western blotting to quantify the dystrophin protein.

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Section: Discussionmentioning

confidence: 99%

Duchenne muscular dystrophy: A immunohistochemical profile and deletion pattern in dystrophin gene in North Indian population

et al. 2017

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“…These findings have also allowed prediction of expected phenotype. Determining carrier status has a considerable significance in estimating the risk in future pregnancies and prenatal testing options to limit the birth of affected individuals [27]. With the possibility that exon skipping techniques may help in some of these children as a therapeutic option, it becomes important that the borders of the deletions are clear and this is possible only by analyzing all the 79 exons in the DMD gene using MLPA technique.…”

Section: Genetics and Patho-physiologymentioning

confidence: 99%

Current Concepts in Dystrophinopathies

Venkataraman

2014

Indian J Pediatr

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Dystrophinopathies comprise a group of hereditary muscle disorders characterized by progressive wasting and weakness of skeletal muscle, as a result of degeneration of muscle fibers, and can be distinguished by the mode of transmission, age at onset and pattern of muscle weakness. The range of phenotypes associated with the region Xp21 has been expanding since identification of the gene in 1987. The mild end of the spectrum includes the phenotype of the muscle cramps with myoglobinuria and isolated quadriceps myopathy, while at the severe end, there are progressive muscle diseases that are classified as Duchenne / Becker muscular dystrophy (DMD/BMD).

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“…Multiplex ligationdependent probe amplification (MLPA) and microarraybased comparative genomic hybridization (aCGH) are superior techniques to multiplex polymerase chain reaction (PCR) for detecting large deletions/duplications in DMD. The MLPA and aCGH allow the identification of a greater number of large deletions, and detect large duplications and provide a better estimation of mutation breakpoints than multiplex PCR (Level of evidence: 1B, Class of Recommendation: A) 12,[17][18][19][20] . Special care should be taken when a single exon deletion is found on MLPA analysis.…”

Section: Diagnosis Confirmationmentioning

confidence: 99%

“…The method of choice will depend on the identified mutation in the index case, generally MLPA or aCGH for large deletions/duplications and Sanger sequencing for point mutations, small deletions or insertions and splice site mutations (Level of evidence: 2B, Class of Recommendation: B) 18,19,20 . It is important to emphasize that multiplex PCR cannot detect heterozygous carriers for large deletions or duplications and, therefore, it is not recommended for carrier detection 12 .…”

Section: Carriers' Detectionmentioning

confidence: 99%

“…The aCGH for DMD is less available than MLPA and the associated costs are generally higher, therefore both an MLPA or an aCGH of DMD are considered first-line tests for DMD diagnosis (Level of evidence: 1B, Class of Recommendation: A) 12,[17][18][19][20] . Southern blot analysis and a multiplex PCR of DMD may also be performed as first-line tests in centers where these are the only available technologies.…”

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Brazilian consensus on Duchenne muscular dystrophy. Part 1: diagnosis, steroid therapy and perspectives

Araujo

Carvalho

Cavalcanti³

et al. 2017

Arq. Neuro-Psiquiatr.

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Significant advances in the understanding and management of Duchenne muscular dystrophy (DMD) took place since international guidelines were published in 2010. Our objective was to provide an evidence-based national consensus statement for multidisciplinary care of DMD in Brazil. A combination of the Delphi technique with a systematic review of studies from 2010 to 2016 was employed to classify evidence levels and grade of recommendations. Our recommendations were divided in two parts. We present Part 1 here, where we describe the guideline methodology and overall disease concepts, and also provide recommendations on diagnosis, steroid therapy and new drug treatment perspectives for DMD. The main recommendations: 1) genetic testing in diagnostic suspicious cases should be the first line for diagnostic confirmation; 2) patients diagnosed with DMD should have steroids prescribed; 3) lack of published results for phase 3 clinical trials hinders, for now, the recommendation to use exon skipping or read-through agents.Keywords: muscular dystrophy, Duchenne; practice guideline; consensus; diagnosis; genetic testing; drug therapy; glucocorticoids; utrophin. RESUMOAvanços na compreensão e no manejo da distrofia muscular de Duchenne (DMD) ocorreram desde a publicação de diretrizes internacionais em 2010. Nosso objetivo foi elaborar um consenso nacional baseado em evidências de cuidado multidisciplinar dos pacientes com DMD no Brasil. Utilizamos a técnica de Delphi combinada com revisão sistemática da literatura de 2010 a 2016 classificando níveis de evidência e graus de recomendação. Nossas recomendações foram divididas em duas partes. Apresentamos aqui a parte 1, descrevendo a metodologia utilizada e conceitos gerais da doença, e fornecemos recomendações sobre diagnóstico, tratamento com corticosteroides e novas perspectivas de tratamentos medicamentosos. As principais recomendações: 1) testes genéticos deveriam ser a primeira linha para confirmação de casos suspeitos; 2) pacientes com diagnóstico de DMD devem receber corticosteroides; 3) por enquanto, a falta de publicações de resultados dos ensaios clínicos de fase 3, dificulta recomendações de uso medicamentos que "saltam exons" ou "passam" por código de parada prematura.

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Identification of deletions and duplications in the Duchenne muscular dystrophy gene and female carrier status in western India using combined methods of multiplex polymerase chain reaction and multiplex ligation-dependent probe amplification

Cited by 11 publications

References 15 publications

Duchenne muscular dystrophy: A immunohistochemical profile and deletion pattern in dystrophin gene in North Indian population

Duchenne muscular dystrophy: A immunohistochemical profile and deletion pattern in dystrophin gene in North Indian population

Current Concepts in Dystrophinopathies

Brazilian consensus on Duchenne muscular dystrophy. Part 1: diagnosis, steroid therapy and perspectives

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