{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,22]],"date-time":"2026-09-22T22:52:19Z","timestamp":1790117539363,"version":"4.0.1"},"reference-count":46,"publisher":"Public Library of Science (PLoS)","issue":"7","license":[{"start":{"date-parts":[[2026,7,21]],"date-time":"2026-07-21T00:00:00Z","timestamp":1784592000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["www.plosone.org"],"crossmark-restriction":false},"short-container-title":["PLoS One"],"abstract":"<jats:sec id=\"sec001\">\n                    <jats:title>Introduction<\/jats:title>\n                    <jats:p>Recently, Molecular diagnosis of Duchenne muscular dystrophy (DMD) and Becker muscular dystrophy (BMD) has become increasingly important in the management of these patients, with techniques such as multiplex ligation-dependent amplification (MLPA) and next-generation sequencing (NGS) coming to the fore. Therefore, this study aims to evaluate the diagnostic accuracy of MLPA, NGS, and the algorithm MLPA-NGS for confirmatory diagnosis of DMD\/BMD.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec id=\"sec002\">\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>We systematically searched databases (PubMed, Embase, Scopus, Cochrane and Web of Science) until July 2025 for studies evaluating the diagnostic accuracy of MLPA and\/or NGS testing in patients with clinical suspicion of DMD, considering multiplex PCR or biopsy as the reference test. A meta-analysis was performed using a random-effects model to estimate the sensitivity, specificity, and detection rate of each test. The QUADAS-2 tool was used to assess the risk of bias and the GRADE criteria were used to identify the certainty of evidence.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec id=\"sec003\">\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>\n                      We included 10 studies (3786 patients) evaluating the use of MLPA and 14 studies (4333 patients) evaluating the use of NGS. For MLPA, the sensitivity was 0.80 (95%CI: 0.76\u20130.84;\n                      <jats:italic>I<\/jats:italic>\n                      <jats:sup>\n                        <jats:italic>2<\/jats:italic>\n                      <\/jats:sup>\n                      : 86%), the specificity was 0.93 (95%CI: 0.87\u20130.96;\n                      <jats:italic>I<\/jats:italic>\n                      <jats:sup>\n                        <jats:italic>2<\/jats:italic>\n                      <\/jats:sup>\n                      : 16%), and AUC 0.90 (CI-95%: 0.89\u20130.92). For NGS, the detection rate was 0.77 (95% CI: 0.61\u20130.87;\n                      <jats:italic>I<\/jats:italic>\n                      <jats:sup>\n                        <jats:italic>2<\/jats:italic>\n                      <\/jats:sup>\n                      : 94%). Furthermore, the detection rate increased to 0.97 (95% CI: 0.94\u20130.99;\n                      <jats:italic>I<\/jats:italic>\n                      <jats:sup>\n                        <jats:italic>2<\/jats:italic>\n                      <\/jats:sup>\n                      : 95%) when NGS was performed after MLPA. We observe a low risk of bias but with very low certainty in the estimations.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec id=\"sec004\">\n                    <jats:title>Conclusions<\/jats:title>\n                    <jats:p>In patients with clinical suspicion of DMD, the MLPA test is very good but with very low certainty. However, in these patients with a negative MLPA, adding the NGS test would allow improve the detection rate. Therefore, the sequential use of these tests could be considered in patients who persist in the clinical suspicion.<\/jats:p>\n                  <\/jats:sec>","DOI":"10.1371\/journal.pone.0345550","type":"journal-article","created":{"date-parts":[[2026,7,21]],"date-time":"2026-07-21T17:28:38Z","timestamp":1784654918000},"page":"e0345550","update-policy":"https:\/\/doi.org\/10.1371\/journal.pone.corrections_policy","source":"Crossref","is-referenced-by-count":1,"title":["Accuracy of the molecular diagnosis of duchenne and becker muscular dystrophy: A systematic review with meta-analysis"],"prefix":"10.1371","volume":"21","author":[{"given":"Fabian A.","family":"Chavez-Ecos","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0076-0985","authenticated-orcid":true,"given":"Carlos","family":"Quispe-Vicu\u00f1a","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Marco","family":"Malaga","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Niels","family":"Pacheco-Barrios","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Andrely","family":"Huerta-Rosario","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Victor","family":"Velasquez-Rimachi","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Peggy","family":"Martinez Esteban","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Carlos","family":"Alva-Diaz","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"340","published-online":{"date-parts":[[2026,7,21]]},"reference":[{"issue":"1","key":"pone.0345550.ref001","doi-asserted-by":"crossref","first-page":"13","DOI":"10.1038\/s41572-021-00248-3","article-title":"Duchenne muscular dystrophy","volume":"7","author":"D Duan","year":"2021","journal-title":"Nat Rev Dis Primers"},{"key":"pone.0345550.ref002","doi-asserted-by":"crossref","DOI":"10.1186\/s13023-020-01430-8","article-title":"Global epidemiology of Duchenne muscular dystrophy: an updated systematic review and meta-analysis","volume":"15","author":"S Crisafulli","year":"2020","journal-title":"Orphanet J Rare 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Systematic Review and Meta-analysis of Diagnostic Test Accuracy Studies: The PRISMA-DTA Statement","volume":"319","author":"and the PRISMA-DTA Group","year":"2018","journal-title":"JAMA"},{"key":"pone.0345550.ref013"},{"key":"pone.0345550.ref014","doi-asserted-by":"crossref","first-page":"529","DOI":"10.7326\/0003-4819-155-8-201110180-00009","article-title":"QUADAS-2: a revised tool for the quality assessment of diagnostic accuracy studies","volume":"155","author":"PF Whiting","year":"2011","journal-title":"Ann Intern Med"},{"key":"pone.0345550.ref015","doi-asserted-by":"crossref","first-page":"129","DOI":"10.1016\/j.jclinepi.2019.12.020","article-title":"GRADE guidelines: 21 part 1. 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Test accuracy: inconsistency, imprecision, publication bias, and other domains for rating the certainty of evidence and presenting it in evidence profiles and summary of findings tables","volume":"122","author":"HJ Sch\u00fcnemann","year":"2020","journal-title":"J Clin Epidemiol"},{"issue":"4","key":"pone.0345550.ref017","first-page":"203","article-title":"Measures of Diagnostic Accuracy: Basic Definitions","volume":"19","author":"A-M \u0160imundi\u0107","year":"2009","journal-title":"EJIFCC"},{"key":"pone.0345550.ref018","doi-asserted-by":"crossref","first-page":"727","DOI":"10.1002\/mus.25396","article-title":"Consecutive analysis of mutation spectrum in the dystrophin gene of 507 Korean boys with Duchenne\/Becker muscular dystrophy in a single center","volume":"55","author":"A Cho","year":"2017","journal-title":"Muscle Nerve"},{"issue":"3","key":"pone.0345550.ref019","doi-asserted-by":"crossref","first-page":"137","DOI":"10.1016\/j.neuarg.2018.05.001","article-title":"An\u00e1lisis del espectro mutacional de la distrofia muscular de Duchenne en un grupo de pacientes colombianos","volume":"10","author":"M Garc\u00eda-Acero","year":"2018","journal-title":"Neurolog\u00eda Argentina"},{"issue":"8","key":"pone.0345550.ref020","doi-asserted-by":"crossref","first-page":"435","DOI":"10.1038\/jhg.2015.43","article-title":"DMD mutation spectrum analysis in 613 Chinese patients with dystrophinopathy","volume":"60","author":"R Guo","year":"2015","journal-title":"J Hum Genet"},{"issue":"1","key":"pone.0345550.ref021","doi-asserted-by":"crossref","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":"X Kong","year":"2019","journal-title":"BMC Med Genet"},{"key":"pone.0345550.ref022","doi-asserted-by":"crossref","first-page":"2177","DOI":"10.1007\/s00415-019-09380-3","article-title":"Mutation pattern in 606 Duchenne muscular dystrophy children with a 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Tomar","year":"2019","journal-title":"Am J Med Genet C Semin Med Genet"},{"issue":"11","key":"pone.0345550.ref028","doi-asserted-by":"crossref","first-page":"856","DOI":"10.3390\/genes10110856","article-title":"Predominance of Dystrophinopathy Genotypes in Mexican Male Patients Presenting as Muscular Dystrophy with A Normal Multiplex Polymerase Chain Reaction DMD Gene Result: A Study Including Targeted Next-Generation Sequencing","volume":"10","author":"MA Alc\u00e1ntara-Ortigoza","year":"2019","journal-title":"Genes (Basel)"},{"issue":"2","key":"pone.0345550.ref029","doi-asserted-by":"crossref","first-page":"199","DOI":"10.1111\/cge.12975","article-title":"Genetic profile of Brazilian patients with dystrophinopathies","volume":"92","author":"PAD de Almeida","year":"2017","journal-title":"Clin Genet"},{"issue":"6","key":"pone.0345550.ref030","doi-asserted-by":"crossref","DOI":"10.1371\/journal.pone.0232654","article-title":"Comprehensive genetic analysis of 961 unrelated Duchenne Muscular Dystrophy patients: Focus on diagnosis, prevention and therapeutic possibilities","volume":"15","author":"SH Kumar","year":"2020","journal-title":"PLoS One"},{"key":"pone.0345550.ref031","doi-asserted-by":"crossref","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":"M Okubo","year":"2016","journal-title":"J Hum Genet"},{"key":"pone.0345550.ref032","doi-asserted-by":"crossref","first-page":"309","DOI":"10.1007\/s12098-017-2455-5","article-title":"Next Generation Sequencing in Diagnosis of MLPA Negative Cases Presenting as Duchenne\/ Becker Muscular Dystrophies","volume":"85","author":"B Singh","year":"2018","journal-title":"Indian J Pediatr"},{"issue":"7","key":"pone.0345550.ref033","doi-asserted-by":"crossref","first-page":"556","DOI":"10.1007\/s13312-019-1553-z","article-title":"Molecular and histopathological 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Several novel variants identified","volume":"30","author":"M Bakhshandeh","year":"2023","journal-title":"eNeurologicalSci"},{"key":"pone.0345550.ref039","doi-asserted-by":"crossref","first-page":"321","DOI":"10.1007\/978-1-4939-8691-0_22","article-title":"Next-Generation Sequencing and Mutational Analysis: Implications for Genes Encoding LINC Complex Proteins","volume":"1840","author":"PL Nagy","year":"2018","journal-title":"Methods Mol Biol"},{"issue":"3","key":"pone.0345550.ref040","doi-asserted-by":"crossref","first-page":"251","DOI":"10.1016\/S1474-4422(18)30024-3","article-title":"Diagnosis and management of Duchenne muscular dystrophy, part 1: diagnosis, and neuromuscular, rehabilitation, endocrine, and gastrointestinal and nutritional management","volume":"17","author":"DJ Birnkrant","year":"2018","journal-title":"Lancet Neurol"},{"key":"pone.0345550.ref041","article-title":"Gu\u00eda de pr\u00e1ctica cl\u00ednica para la detecci\u00f3n temprana, atenci\u00f3n integral, seguimiento y rehabilitaci\u00f3n de pacientes con diagn\u00f3stico de distrofia muscular","author":"Sistema General de Seguridad Social en Salud","year":"2014"},{"issue":"3","key":"pone.0345550.ref042","doi-asserted-by":"crossref","first-page":"3245","DOI":"10.3390\/ijms13033245","article-title":"Use of the MLPA assay in the molecular diagnosis of gene copy number alterations in human genetic diseases","volume":"13","author":"L Stuppia","year":"2012","journal-title":"Int J Mol Sci"},{"issue":"1","key":"pone.0345550.ref043","doi-asserted-by":"crossref","first-page":"91","DOI":"10.4103\/0971-6866.96667","article-title":"Utility of MLPA in mutation analysis and carrier detection for Duchenne muscular dystrophy","volume":"18","author":"PK Verma","year":"2012","journal-title":"Indian J Hum Genet"},{"issue":"9","key":"pone.0345550.ref044","article-title":"MLPA followed by target-NGS to detect mutations in the dystrophin gene of Peruvian patients suspected of 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