Sensitivity and Frequencies of Dystrophin Gene Mutations in Thai DMD/BMD Patients As Detected by Multiplex PCR (original) (raw)
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BMC Research Notes, 2019
Objective: Duchenne/Becker muscular dystrophy (DMD/BMD) is the most common genetic neuromuscular disease in children, resulting from a defect in the DMD gene located on Xp21.2. The new emerging treatment using exon skipping strategy is tailored to specific mutations, thus molecular diagnostics are particularly important. This study aimed to detect the DMD gene deletion in Indonesian DMD/BMD patients and analyze the potential amenability by exon skipping therapy. Results: Thirty-four male patients were enrolled in this study, 23 of them (67.6%) underwent muscle biopsy and showed the absence or partially expressed dystrophin protein in immunohistochemistry staining. All patients had very high serum CK levels (10.529 ± 9.97 IU/L). Multiplex PCR revealed the DMD gene deletions in 15 (44.1%) cases. Seventy-eight percent of deletions were clustered in the hot-spot region of exon 43 to 52. Furthermore, seven (20.5%) patients were potentially amenable to exon skipping treatment. Therefore, multiplex PCR is one feasible method to detect DMD gene deletion in Indonesian DMD/BMD patients that can further determine the potential amenability of exon skipping therapy. In addition, this study is the first report of DMD gene deletion analysis in Indonesia.
Mutation analysis in Saudi Duchenne and Becker muscular dystrophy patients using multiplex PCR
Archives of Medical …, 2008
A b s t r a c t I In nt tr ro od du uc ct ti io on n: : In Saudi Arabia, only limited work has been reported on the mutation patterns of Duchenne muscular dystrophy (DMD) and Becker muscular dystrophy (BMD). This study looks at the spectrum of deletions in the 'hot spot' regions of the DMD gene in Saudi DMD/BMD patients using an enhanced multiplex PCR technique. M Ma at te er ri ia al l a an nd d m me et th ho od ds s: : Twenty-six exons of the DMD gene were analyzed, in eight unrelated DMD/BMD cases aged 4-19 years, using four different multiplex PCR sets. Each multiplex PCR set amplified a total of six or seven exons. Normal controls were included for validation. R Re es su ul lt ts s: : Using an optimized multiplex PCR method, 5 out of 8 DMD/BMD patients showed deletions, while the remaining three had no deletions in regions analyzed. Set 1 detected no deletions in any of the patients, whereas each of sets 2, 3 and 4 detected two, four and three deletions respectively. All of these mutations were located in the distal 'hot spot' region. No deletions were detected in the proximal 'hot spot' region. The normal control samples showed no deletions in any of the 26 exons tested. C Co on nc cl lu us si io on ns s: : In this study, multiplex PCR technology was utilized to demonstrate the frequency of the most commonly found deletions in a limited group of Saudi DMD/BMD patients. The overall distribution of deletion mutations in the distal 'hot spot' region was in accordance with DMD/BMD cases investigated elsewhere. The study also serves as a good starting point for further investigations into the genetic aspects of the Saudi DMD/BMD population. K Ke ey y w wo or rd ds s: : dystrophin, Saudi DMD, BMD, deletions, multiplex PCR, DMD gene. C Co or rr re es sp po on nd di in ng g a au ut th ho or r: :
Analysis of Dystrophin Gene Deletions by Multiplex PCR in Moroccan Patients
Journal of Biomedicine and Biotechnology, 2002
Duchenne and Becker muscular dystrophy (DMD and BMD) are X-linked diseases resulting from a defect in the dystrophin gene located on Xp21. DMD is the most frequent neuromuscular disease in humans (1/3500 male newborn). Deletions in the dystrophin gene represent 65% of mutations in DMD/BMD patients. We have analyzed DNA from 72 Moroccan patients with DMD/BMD using the multiplex polymerase chain reaction (PCR) to screen for exon deletions within the dystrophin gene, and to estimate the frequency of these abnormalities. We found dystrophin gene deletions in 37 cases. Therefore the frequency in Moroccan DMD/BMD patients is about 51.3%. All deletions were clustered in the two known hot-spots regions, and in 81% of cases deletions were detected in the region from exon 43 to exon 52. These findings are comparable to those reported in other studies. It is important to note that in our population, we can first search for deletions of DMD gene in the most frequently deleted exons determined b...
Screening of Dystrophin Gene Deletions in Egyptian Patients with DMD/BMD Muscular Dystrophies
Disease Markers, 2000
Duchenne muscular dystrophy (DMD) and Becker muscular dystrophy (BMD) are allelic disorders caused by mutations within the dystrophin gene. Our study has identified 100 Egyptian families collected from the Human Genetics Clinic, National Research Center, Cairo. All cases were subjected to complete clinical evaluation pedigree analysis, electromyography studies, estimation of serum creatine phosphokinase enzyme (CPK) levels and DNA analysis. Multiplex PCR using 18 pairs of specific primers were used for screening of deletion mutations within the dystrophin gene. A frequency of 55% of deletions were found among the families. Sixty per cent of detected deletions involved multiple exons spanning the major or the minor hot spot of the dystrophin gene. The remainder 40% involved single exon deletions, which mainly involved exon 45. Comparing these findings with frequencies of other countries it was found that our figures fall within the reported range of 40%-60% for deletions. The distribution of deletions in our study and other different studies was variable and specific ethnic differences do not apparently account for specific deletions. In addition this study concluded that employment of the 18 exon analysis is a cost effective and a highly accurate (97% detection rate) method to be considered when planning to launch a nationwide program.
Clinical Genetics, 2008
Fifty unrelated Japanese patients with Duchenne and Becker muscular dystrophy (DMD and BMD) have been studied through use of the dystrophin cDNA probes. The 14-kb dystrophin cDNA was subdivided into six subclones, and Hind III-digested DNAs were analyzed by Southern blotting. Of 50 unrelated patients, 20 showed a deletion of one or several of the exon-containing Hind III fragments (40.0%). These corresponded to 50% (11/22) of BMD patients and 32.1% (9/28) of DMD patients, and the position and extent of deletions were mapped and proven to be more heterogeneous in DMD than in BMD. Both ends of deletions detected by probe 1-2a were common to all six BMD patients, and the 5' ends of deletions in probe 5b-7 were also common to four BMD patients. The phenotypic-specific deletion in Japanese BMD patients existed in the 5' end of the DMD gene, although an apparently similar deletion produced a wide range of clinical courses (BMD phenotype). Three out of eight females in DMD/BMD families were diagnosed as carriers through use of the junctional fragment and dosage analyses of dystrophin cDNA.
Deneysel tıp dergisi, 2017
Background: Duchenne/Becker muscular dystrophy (DMD/BMD) is an X-linked recessive disease results from mutations in the dystrophin gene. We established the deletion pattern profile in unrelated DMD/BMD patients using multiplex PCR (M-PCR). Methods: During 1998-2015, 1,385 unrelated Deletion analysis in the dystrophin(DMD) gene was performed.Results: Of all patients admitted, 42.6% deletion carriers (n=589) were detected, of which 180 (80.3 %)were carrying single exon deletions and 409 (14.8 %) multiple exon deletions. Deletions covering the major hotspot region were 80.3 %, the minor region 14.8% and 2.4% covered both regions. The mean age of diagnosis of patients with out-of-frame deletions (7.27 year) was notably lower than the cases with in frame deletions (17.54 year). No single exon 4 deletion was detected.Conclusions: When the known deletion hotspots are considered, the study population showed a similar deletion pattern with other populations. The mean age of patients with out-of-frame deletions were lower than mean age of those with in-frame deletions, in concordance with the reading frame hypothesis. Strikingly, no single exon 4 deletion was found, supporting the hypothesis that absence of it might have no functional consequences.
Screening of deletions and RFLP analysis in Turkish DMD/BMD families by PCR
Clinical Genetics, 2008
Gökgöz N, Kuseyri F, TopaloǦlu H, Yüksel-Apak M, Kirdar B. Screening of deletions and RFLP analysis in Turkish DMD/BMD families by PCR.Clin Genet 1993: 43: 261–266. © Munksgaard, 1993We have screened 76 DMD and 5 BMD patients for deletions, using two separate Multiplex gene amplification systems. The use of both systems together revealed deletions in 52% of the cases in the Turkish population. The majority of these deletions (33/37) were found to be localized within the central region of the dystrophin gene. The remaining deletions were mapped to the proximal hotspot. Deletion end-points were identified by PCR and/or by Southern blot analysis with cDNA probes, and exceptions to the Open Reading Frame (ORF) hypothesis are discussed. PCR-based techniques to screen the pER.T87.15/XmnI, pERT87.15/BamHI, and pERT87.8/TaqI polymorphisms were used for linkage analysis in the Turkish DMD/BMD families, and approximately 70% of the mothers at risk were found to be informative for at least one of these polymorphisms studied.