Detecting exon deletions and duplications of the DMD gene using Multiplex Ligation-dependent Probe Amplification (MLPA)

Lai, Kent; Lo, Ivan F.M.; Tong, Tony M F; Cheng, L. Y. L.; Lam, Stephen

doi:10.1016/j.clinbiochem.2005.11.019

Cited by 66 publications

(45 citation statements)

References 26 publications

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“…Compared with existing methods, such as QF-PCR assays (quantitative multiplex-PCR of short fluorescent fragments, quantitative fluorescent multiplex-PCR, and semiquantitative fluorescent-PCR, developed to screen for deletions and duplications of all exons of DMD or CFTR gene) [White et al, 2002;Gatta et al, 2005;Lalic et al, 2005;Lai et al, 2006], the CGH array assay described in this study appears to be very promising to address diagnostic and research issues. First, as shown here, the CGH array enables detection of exonic deletions and duplications while providing the possibility to confine intronic breakpoints involved in the rearrangements.…”

Section: Discussionmentioning

confidence: 99%

Detection of exonic copy-number changes using a highly efficient oligonucleotide-based comparative genomic hybridization-array method

et al. 2008

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For the Focus Section on Array-CGHGenomic copy-number variations (CNVs) involving large DNA segments are known to cause many genetic disorders. Depending on the changes, they are predicted to lead either to decreased or an increased gene expression. However, the ability to detect smaller exonic copy-number changes has not been explored. Here we describe a new oligonucleotide-based comparative genomic hybridization (CGH)-array approach for highthroughput detection of exonic deletions or duplications and its application to deletion/duplication analyses of the genes encoding CFTR, six sarcoglycans (SGCA, SGCB, SGCG, SGCD, SGCE, and SGCZ), and DMD. In this work we show the successful development of an array format containing 158 exons that collectively span eight genes and its clinical application for the rapid screening of deletions and duplications in a diagnostic setting. We have analyzed a series of 35 DNA samples from patients affected with cystic fibrosis (CF), Duchenne and Becker muscular dystrophies (DMD/BMD), or sarcoglycanopathies, and have characterized exonic copy-number changes that have been validated with other methods. Interestingly, even heterozygous deletions and duplications of only one exon, as well as mosaic deletions, were detected by this CGH approach. Our results showed that the resolution is very high, as abnormalities of about 1.5-2 kb could be detected. Since this approach is completely scalable, this new molecular tool will allow the screening of combinations of genes involved in a particular group of clinically and genetically heterogeneous disorders such as mental retardation, muscular dystrophies and brain malformations. Hum Mutat 29(9), [1083][1084][1085][1086][1087][1088][1089][1090] 2008.

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Section: Discussionmentioning

confidence: 99%

Detection of exonic copy-number changes using a highly efficient oligonucleotide-based comparative genomic hybridization-array method

et al. 2008

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“…Two control samples and a positive control with a deletion of exons 1-8 (17) and a negative control were included in the assay. Dosage quotients were calculated using previously described methods (18). Real-time PCR quantification of the mutant allele.…”

Section: Methodsmentioning

confidence: 99%

An ABCC8 Gene Mutation and Mosaic Uniparental Isodisomy Resulting in Atypical Diffuse Congenital Hyperinsulinism

et al. 2008

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OBJECTIVE-Congenital hyperinsulinism (CHI) may be due to diffuse or focal pancreatic disease. The diffuse form is associated with an increase in the size of ␤-cell nuclei throughout the whole of the pancreas and most commonly results from recessive ATP-sensitive K ϩ channel (K ATP channel) mutations. Focal lesions are the consequence of somatic uniparental disomy for a paternally inherited K ATP channel mutation with enlargement of the ␤-cell nuclei confined to the focal lesion. Some "atypical" cases defy classification and show pancreatic ␤-cell nuclear enlargement confined to discrete regions of the pancreas. We investigated an atypical case with normal morphology within the tail of the pancreas but occasional enlarged endocrine nuclei in parts of the body and head. RESEARCH DESIGN AND METHODS-The KCNJ11 andABCC8 genes encoding the K ATP channel subunits and microsatellite markers on chromosome 11 were analyzed in DNA samples from the patient and her parents.RESULTS-A mosaic ABCC8 nonsense mutation (Q54X) was identified in the proband. The paternally inherited mutation was present at 90% in lymphocytes and 50% in normal pancreatic sections but between 64 and 74% in abnormal sections. Microsatellite analysis showed mosaic interstitial paternal uniparental isodisomy (UPD) for chromosome 11p15.1.CONCLUSIONS-We report a novel genetic mechanism to explain atypical histological diffuse forms of CHI due to mosaic UPD in patients with dominantly inherited ABCC8 (or KCNJ11) gene mutations.

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“…A number of studies have found MLPA assay to be highly sensitive diagnostic assay to identify mutation in DMD and BMD patients [9]. In the study to evaluate the efficacy of MLPA technique in comparison with the traditional multiplex PCR assay in detection of exon deletions and duplications of the DMD gene by Lai et al, MLPA was able to detect all the known deletions and duplications; it detected four additional mutations that had been missed by multiplex PCR [10]. In a study in China amongst 70 DMD/BMD patients, MLPA detected exonic deletions in 42 patients (60%), exonic duplications in 7 patients (10%) and 21 patients (30%) showed normal results [11].…”

Section: Discussionmentioning

confidence: 99%

Multiplex Ligation Dependent Probe Amplification Based Mutation Analysis of Dystrophin Gene in Nepalese Patients with Duchenne Muscular Dystrophy

Shrestha

Khatiwada³

et al. 2016

Nepal J Biotechnol

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Duchenne muscular dystrophy (DMD) is X-linked recessive neuromuscular disorders caused due to mutation in dystrophin gene, leading to progressive muscle weakness. This study was done to identify mutation in dystrophin gene in Nepalese patients with DMD using Multiplex Ligation Dependent Probe Amplification (MLPA) assay in Nepal. Twenty one patients from different regions of Nepal, who were clinically diagnosed as DMD were enrolled in the study. Peripheral blood samples were collected in EDTA vials, gDNA was extracted, and deletion mutation in the dystrophin gene was analysed using Multiplex Ligation Dependent Probe Amplification (MLPA) assay. Exon deletion mutation in the dystrophin gene was observed in 14 (66.6%) out of 21 DMD cases. The most common exon deletion was observed and confined in exon 7-14 and 45-53 of dystrophin gene. The location of deletion in dystrophin gene is apparently non-random with a preponderance found in the hot spot regions. Use of MLPA is useful in detecting copy number changes in DMD proband and suspected carriers in Nepal.

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Detecting exon deletions and duplications of the DMD gene using Multiplex Ligation-dependent Probe Amplification (MLPA)

Cited by 66 publications

References 26 publications

Detection of exonic copy-number changes using a highly efficient oligonucleotide-based comparative genomic hybridization-array method

Detection of exonic copy-number changes using a highly efficient oligonucleotide-based comparative genomic hybridization-array method

An ABCC8 Gene Mutation and Mosaic Uniparental Isodisomy Resulting in Atypical Diffuse Congenital Hyperinsulinism

Multiplex Ligation Dependent Probe Amplification Based Mutation Analysis of Dystrophin Gene in Nepalese Patients with Duchenne Muscular Dystrophy

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