{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,7,30]],"date-time":"2025-07-30T15:54:57Z","timestamp":1753890897758,"version":"3.41.2"},"reference-count":39,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2021,12,3]],"date-time":"2021-12-03T00:00:00Z","timestamp":1638489600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Genet."],"abstract":"<jats:p>Muscular Dystrophies (MDs) are a group of inherited diseases and heterogeneous in nature. To date, 40 different genes have been reported for the occurrence and\/or progression of MDs. This study was conducted to demonstrate the application of next-generation sequencing (NGS) in developing a time-saving and cost-effective diagnostic method to detect single nucleotide variants (SNVs) and copy number variants (CNVs) in a single test. A total of 123 cases clinically suspected of MD were enrolled in this study. Amplicon panel-based diagnosis was carried out for 102 (DMD\/BMD) cases and the results were further screened using multiplex ligation-dependent probe amplification (MLPA). Whilst in the case of LGMD (N = 19) and UMD (N = 2), only NGS panel-based analysis was carried out. We identified the large deletions in 74.50% (76\/102) of the cases screened with query DMD or BMD. Further, the large deletion in <jats:italic>CAPN3<\/jats:italic> gene (N = 3) and known SNV mutations (N = 4) were identified in LGMD patients. Together, the total diagnosis rate for this amplicon panel was 70.73% (87\/123) which demonstrated the utility of panel-based diagnosis for high throughput, affordable, and time-saving diagnostic strategy. Collectively, present study demonstrates that the panel based NGS sequencing could be superior over to MLPA.<\/jats:p>","DOI":"10.3389\/fgene.2021.770350","type":"journal-article","created":{"date-parts":[[2021,12,3]],"date-time":"2021-12-03T11:49:23Z","timestamp":1638532163000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":2,"title":["Molecular Diagnosis of Muscular Dystrophy Patients in Western Indian Population: A Comprehensive Mutation Analysis Using Amplicon Sequencing"],"prefix":"10.3389","volume":"12","author":[{"given":"Komal M.","family":"Patel","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Arpan D.","family":"Bhatt","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Krati","family":"Shah","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Bhargav N.","family":"Waghela","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ramesh J.","family":"Pandit","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Harsh","family":"Sheth","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Chaitanya G.","family":"Joshi","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Madhvi N.","family":"Joshi","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1965","published-online":{"date-parts":[[2021,12,3]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","first-page":"135","DOI":"10.1002\/mus.20586","article-title":"Entries in the Leiden Duchenne Muscular Dystrophy Mutation Database: an Overview of Mutation Types and Paradoxical Cases that Confirm the reading-frame Rule","volume":"34","author":"Aartsma-Rus","year":"2006","journal-title":"Muscle Nerve."},{"key":"B2","doi-asserted-by":"publisher","first-page":"145","DOI":"10.1136\/jmedgenet-2015-103387","article-title":"The Importance of Genetic Diagnosis for Duchenne Muscular Dystrophy","volume":"53","author":"Aartsma-Rus","year":"2016","journal-title":"J. Med. Genet."},{"key":"B3","doi-asserted-by":"publisher","first-page":"731","DOI":"10.1016\/j.jmoldx.2016.05.003","article-title":"Implementation of a Reliable Next-Generation Sequencing Strategy for Molecular Diagnosis of Dystrophinopathies","volume":"18","author":"Alame","year":"2016","journal-title":"J. Mol. Diagn."},{"key":"B4","doi-asserted-by":"publisher","first-page":"693","DOI":"10.1080\/14737175.2018.1508997","article-title":"An Update on Diagnostic Options and Considerations in Limb-Girdle Dystrophies","volume":"18","author":"Angelini","year":"2018","journal-title":"Expert Rev. Neurotherapeutics"},{"key":"B5","doi-asserted-by":"publisher","first-page":"282","DOI":"10.4103\/ijmr.IJMR_290_18","article-title":"Targeted Sequencing of the DMD Locus: A Comprehensive Diagnostic Tool for All Mutations","volume":"150","author":"Aravind","year":"2019","journal-title":"Indian J. Med. Res."},{"volume-title":"Creatine Phosphokinase","year":"2020","author":"Aujla","key":"B6"},{"key":"B7","first-page":"122","article-title":"Genetic Diagnosis as a Tool for Personalized Treatment of Duchenne Muscular Dystrophy","volume":"35","author":"Bello","year":"2016","journal-title":"Acta Myol."},{"key":"B8","doi-asserted-by":"publisher","DOI":"10.7860\/jcdr\/2019\/40923.12812","article-title":"A Case of Limb Girdle Muscular Dystrophy Type 2A from India: Copy Number Variation Analysis Using Targeted Amplicon Sequencing","volume":"13","author":"Bhatt","year":"2019","journal-title":"J. Clin. Diagn. Res."},{"key":"B9","doi-asserted-by":"publisher","first-page":"779","DOI":"10.7196\/samj.8257","article-title":"Duchenne Muscular Dystrophy: High-Resolution Melting Curve Analysis as an Affordable Diagnostic Mutation Scanning Tool in a South African Cohort","volume":"104","author":"Esterhuizen","year":"2014","journal-title":"S Afr. Med. J."},{"key":"B10","doi-asserted-by":"crossref","DOI":"10.5772\/intechopen.85339","volume-title":"Clinical and Molecular Diagnosis in Muscular Dystrophies","author":"Gaina","year":"2019"},{"key":"B11","doi-asserted-by":"publisher","first-page":"1919","DOI":"10.1007\/s00415-019-09358-1","article-title":"Multi-gene Testing in Neurological Disorders Showed an Improved Diagnostic Yield: Data from over 1000 Indian Patients","volume":"266","author":"Ganapathy","year":"2019","journal-title":"J. Neurol."},{"key":"B12","doi-asserted-by":"publisher","first-page":"1954","DOI":"10.1212\/con.0000000000000406","article-title":"The Limb-Girdle Muscular Dystrophies and the Dystrophinopathies","volume":"22","author":"Iyadurai","year":"2016","journal-title":"CONTINUUM: Lifelong Learn. Neurol."},{"key":"B13","doi-asserted-by":"publisher","first-page":"495","DOI":"10.1007\/s12098-020-03286-z","article-title":"Mutation Spectrum of Dystrophinopathies in India: Implications for Therapy","volume":"87","author":"Kohli","year":"2020","journal-title":"Indian J. Pediatr."},{"key":"B14","doi-asserted-by":"publisher","first-page":"139","DOI":"10.1186\/s12881-019-0873-0","article-title":"Genetic Analysis of 1051 Chinese Families with Duchenne\/Becker Muscular Dystrophy","volume":"20","author":"Kong","year":"2019","journal-title":"BMC Med. Genet."},{"key":"B15","doi-asserted-by":"publisher","first-page":"128","DOI":"10.1016\/j.bbadis.2006.07.002","article-title":"Molecular and Cellular Basis of Calpainopathy (Limb Girdle Muscular Dystrophy Type 2A)","volume":"1772","author":"Kramerova","year":"2007","journal-title":"Biochim. Biophys. Acta (Bba) - Mol. Basis Dis."},{"key":"B16","doi-asserted-by":"publisher","first-page":"D980","DOI":"10.1093\/nar\/gkt1113","article-title":"ClinVar: Public Archive of Relationships Among Sequence Variation and Human Phenotype","volume":"42","author":"Landrum","year":"2014","journal-title":"Nucl. Acids Res."},{"key":"B17","doi-asserted-by":"publisher","first-page":"731","DOI":"10.1136\/jmedgenet-2011-100133","article-title":"Genetic Diagnosis of Duchenne and Becker Muscular Dystrophy Using Next-Generation Sequencing Technology: Comprehensive Mutational Search in a Single Platform","volume":"48","author":"Lim","year":"2011","journal-title":"J. Med. Genet."},{"key":"B18","doi-asserted-by":"publisher","first-page":"465","DOI":"10.1017\/s0317167100011896","article-title":"A Population-Based Study of Dystrophin Mutations in Canada","volume":"38","author":"Mah","year":"2011","journal-title":"Can. J. Neurol. Sci."},{"key":"B19","doi-asserted-by":"publisher","first-page":"e0197205","DOI":"10.1371\/journal.pone.0197205","article-title":"Mutation Spectrum Analysis of Duchenne\/Becker Muscular Dystrophy in 68 Families in Kuwait: The Era of Personalized Medicine","volume":"13","author":"Mohammed","year":"2018","journal-title":"PloS one"},{"key":"B20","doi-asserted-by":"publisher","first-page":"S7","DOI":"10.3233\/jnd-150105","article-title":"The Classification, Natural History and Treatment of the Limb Girdle Muscular Dystrophies","volume":"2","author":"Murphy","year":"2015","journal-title":"Jnd"},{"key":"B21","doi-asserted-by":"publisher","first-page":"1574","DOI":"10.1002\/acn3.649","article-title":"Genetic Landscape and Novel Disease Mechanisms from a largeLGMDcohort of 4656 Patients","volume":"5","author":"Nallamilli","year":"2018","journal-title":"Ann. Clin. Transl Neurol."},{"key":"B23","doi-asserted-by":"publisher","first-page":"483","DOI":"10.1038\/jhg.2016.7","article-title":"Genetic Diagnosis of Duchenne\/Becker Muscular Dystrophy Using Next-Generation Sequencing: Validation Analysis of DMD Mutations","volume":"61","author":"Okubo","year":"2016","journal-title":"J. Hum. Genet."},{"key":"B24","doi-asserted-by":"publisher","first-page":"413","DOI":"10.1007\/s13353-011-0057-x","article-title":"Sequencing Technologies and Genome Sequencing","volume":"52","author":"Pareek","year":"2011","journal-title":"J. Appl. Genet."},{"key":"B25","first-page":"1993","article-title":"Limb-Girdle Muscular Dystrophy Overview \u2013 RETIRED CHAPTER, FOR HISTORICAL REFERENCE ONLY","volume-title":"GeneReviews\u00ae [Internet]","author":"Pegoraro","year":"2012"},{"key":"B26","doi-asserted-by":"publisher","first-page":"2177","DOI":"10.1007\/s00415-019-09380-3","article-title":"Mutation Pattern in 606 Duchenne Muscular Dystrophy Children with a Comparison between Familial and Non-familial Forms: a Study in an Indian Large Single-center Cohort","volume":"266","author":"Polavarapu","year":"2019","journal-title":"J. Neurol."},{"key":"B27","doi-asserted-by":"publisher","first-page":"405","DOI":"10.1038\/gim.2015.30","article-title":"Standards and Guidelines for the Interpretation of Sequence Variants: a Joint Consensus Recommendation of the American College of Medical Genetics and Genomics and the Association for Molecular Pathology","volume":"17","author":"Richards","year":"2015","journal-title":"Genet. Med."},{"key":"B28","doi-asserted-by":"publisher","first-page":"24","DOI":"10.1038\/nbt.1754","article-title":"Integrative Genomics Viewer","volume":"29","author":"Robinson","year":"2011","journal-title":"Nat. Biotechnol."},{"key":"B29","doi-asserted-by":"publisher","first-page":"111","DOI":"10.3390\/jpm10030111","article-title":"Advances in Genetic Characterization and Genotype-Phenotype Correlation of Duchenne and Becker Muscular Dystrophy in the Personalized Medicine Era","volume":"10","author":"Sheikh","year":"2020","journal-title":"Jpm"},{"key":"B30","doi-asserted-by":"publisher","first-page":"702","DOI":"10.1016\/j.nmd.2018.05.007","article-title":"229th ENMC International Workshop: Limb Girdle Muscular Dystrophies - Nomenclature and Reformed Classification Naarden, the Netherlands, 17-19 March 2017","volume":"28","author":"Straub","year":"2018","journal-title":"Neuromuscul. Disord."},{"key":"B31","doi-asserted-by":"publisher","first-page":"1648","DOI":"10.1212\/wnl.44.9.1648","article-title":"Amino-terminal Deletion of 53% of Dystrophin Results in an Intermediate Duchenne-Becker Muscular Dystrophy Phenotype","volume":"44","author":"Takeshima","year":"1994","journal-title":"Neurology"},{"key":"B32","doi-asserted-by":"publisher","first-page":"e53083","DOI":"10.1371\/journal.pone.0053083","article-title":"Comprehensive Mutation Analysis for Congenital Muscular Dystrophy: a Clinical PCR-Based Enrichment and Next-Generation Sequencing Panel","volume":"8","author":"Valencia","year":"2013","journal-title":"PloS one"},{"key":"B33","doi-asserted-by":"publisher","first-page":"814","DOI":"10.3389\/fphar.2019.00814","article-title":"Molecular Genetics Analysis of 70 Chinese Families with Muscular Dystrophy Using Multiplex Ligation-dependent Probe Amplification and Next-Generation Sequencing","volume":"10","author":"Wang","year":"2019","journal-title":"Front. Pharmacol."},{"key":"B34","doi-asserted-by":"publisher","first-page":"1013","DOI":"10.1007\/s00438-014-0847-z","article-title":"Whole Dystrophin Gene Analysis by Next-Generation Sequencing: a Comprehensive Genetic Diagnosis of Duchenne and Becker Muscular Dystrophy","volume":"289","author":"Wang","year":"2014","journal-title":"Mol. Genet. Genomics"},{"key":"B35","doi-asserted-by":"publisher","first-page":"110","DOI":"10.1038\/ejhg.2013.82","article-title":"Targeted Next-Generation Sequencing as a Comprehensive Test for Patients with and Female Carriers of DMD\/BMD: a Multi-Population Diagnostic Study","volume":"22","author":"Wei","year":"2014","journal-title":"Eur. J. Hum. Genet."},{"key":"B36","doi-asserted-by":"publisher","first-page":"1535","DOI":"10.1212\/01.con.0000440659.41675.8b","article-title":"The Muscular Dystrophies","volume":"19","author":"Wicklund","year":"2013","journal-title":"CONTINUUM: Lifelong Learn. Neurol."},{"key":"B37","doi-asserted-by":"publisher","first-page":"8","DOI":"10.1186\/s13039-017-0301-0","article-title":"Identification of a Novel DMD Duplication Identified by a Combination of MLPA and Targeted Exome Sequencing","volume":"10","author":"Wu","year":"2017","journal-title":"Mol. Cytogenet."},{"key":"B38","doi-asserted-by":"publisher","first-page":"29","DOI":"10.1186\/1471-2350-14-29","article-title":"MLPA-based Genotype-Phenotype Analysis in 1053 Chinese Patients with DMD\/BMD","volume":"14","author":"Yang","year":"2013","journal-title":"BMC Med. Genet."},{"key":"B39","doi-asserted-by":"publisher","first-page":"66","DOI":"10.1016\/j.cca.2019.01.014","article-title":"Molecular Genetic Testing and Diagnosis Strategies for Dystrophinopathies in the Era of Next Generation Sequencing","volume":"491","author":"Zhang","year":"","journal-title":"Clinica Chim. Acta"},{"key":"B40","doi-asserted-by":"publisher","first-page":"e622","DOI":"10.1002\/mgg3.622","article-title":"Application Whole Exome Sequencing for the Clinical Molecular Diagnosis of Patients with Duchenne Muscular Dystrophy; Identification of Four Novel Nonsense Mutations in Four Unrelated Chinese DMD Patients","volume":"7","author":"Zhang","year":"","journal-title":"Mol. Genet. Genomic Med."}],"container-title":["Frontiers in Genetics"],"original-title":[],"link":[{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fgene.2021.770350\/full","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2021,12,3]],"date-time":"2021-12-03T11:49:32Z","timestamp":1638532172000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fgene.2021.770350\/full"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2021,12,3]]},"references-count":39,"alternative-id":["10.3389\/fgene.2021.770350"],"URL":"https:\/\/doi.org\/10.3389\/fgene.2021.770350","relation":{},"ISSN":["1664-8021"],"issn-type":[{"type":"electronic","value":"1664-8021"}],"subject":[],"published":{"date-parts":[[2021,12,3]]},"article-number":"770350"}}