{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,4,7]],"date-time":"2026-04-07T22:01:45Z","timestamp":1775599305887,"version":"3.50.1"},"reference-count":36,"publisher":"Springer Science and Business Media LLC","issue":"1","license":[{"start":{"date-parts":[[2021,2,17]],"date-time":"2021-02-17T00:00:00Z","timestamp":1613520000000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"},{"start":{"date-parts":[[2021,2,17]],"date-time":"2021-02-17T00:00:00Z","timestamp":1613520000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"}],"funder":[{"name":"Special Foundation for High\u2010level Talents of Guangdong","award":["2016TX03R171"],"award-info":[{"award-number":["2016TX03R171"]}]}],"content-domain":{"domain":["link.springer.com"],"crossmark-restriction":false},"short-container-title":["Sci Rep"],"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>Hearing loss is one of the most common birth disorders in humans, with an estimated prevalence of 1\u20133 in every 1000 newborns. This study investigates the molecular etiology of a hearing loss cohort using a stepwise strategy to effectively diagnose patients and address the challenges posed by the genetic heterogeneity and variable mutation spectrum of hearing loss. In order to target known pathogenic variants, multiplex PCR plus next-generation sequencing was applied in the first step; patients which did not receive a diagnosis from this were further referred for exome sequencing. A total of 92 unrelated patients with nonsyndromic hearing loss were enrolled in the study. In total, 64% (59\/92) of the patients were molecularly diagnosed, 44 of them in the first step by multiplex PCR plus sequencing. Exome sequencing resulted in eleven diagnoses (23%, 11\/48) and four probable diagnoses (8%, 4\/48) among the 48 patients who were not diagnosed in the first step. The rate of secondary findings from exome sequencing in our cohort was 3% (2\/58). This research presents a molecular diagnosis spectrum of 92 non-syndromic hearing loss patients and demonstrates the benefits of using a stepwise diagnostic approach in the genetic testing of nonsyndromic hearing loss.<\/jats:p>","DOI":"10.1038\/s41598-021-83493-6","type":"journal-article","created":{"date-parts":[[2021,2,19]],"date-time":"2021-02-19T11:46:37Z","timestamp":1613735197000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":10,"title":["Molecular diagnosis of non-syndromic hearing loss patients using a stepwise approach"],"prefix":"10.1038","volume":"11","author":[{"given":"Jing","family":"Wang","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jiale","family":"Xiang","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Lisha","family":"Chen","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Hongyu","family":"Luo","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Xiuhua","family":"Xu","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Nan","family":"Li","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Chunming","family":"Cui","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jingjing","family":"Xu","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Nana","family":"Song","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jiguang","family":"Peng","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Zhiyu","family":"Peng","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"297","published-online":{"date-parts":[[2021,2,17]]},"reference":[{"issue":"20","key":"83493_CR1","doi-asserted-by":"publisher","first-page":"2151","DOI":"10.1056\/NEJMra050700","volume":"354","author":"CCN Morton","year":"2006","unstructured":"Morton, C. C. N. & Walter, E. Newborn hearing screening\u2014A silent revolution. N. Engl. J. Med. 354(20), 2151\u20132164 (2006).","journal-title":"N. Engl. J. Med."},{"issue":"5","key":"83493_CR2","doi-asserted-by":"publisher","first-page":"260","DOI":"10.1016\/j.mcp.2015.03.008","volume":"29","author":"B Vona","year":"2015","unstructured":"Vona, B., Nanda, I., Hofrichter, M. A., Shehata-Dieler, W. & Haaf, T. Non-syndromic hearing loss gene identification: A brief history and glimpse into the future. Mol. Cell Probes 29(5), 260\u2013270 (2015).","journal-title":"Mol. Cell Probes"},{"key":"83493_CR3","unstructured":"Van Camp, G. & Smith. R. Hereditary hearing loss homepage. https:\/\/hereditaryhearingloss.org. Accessed: August 2019."},{"key":"83493_CR4","doi-asserted-by":"publisher","first-page":"2744","DOI":"10.1038\/s41436-019-0594-y","volume":"21","author":"M Li","year":"2019","unstructured":"Li, M. et al. Extrusion pump ABCC1 was first linked with nonsyndromic hearing loss in humans by stepwise genetic analysis. Genet. Med. 21, 2744\u20132754 (2019).","journal-title":"Genet. Med."},{"issue":"1","key":"83493_CR5","doi-asserted-by":"publisher","first-page":"16094","DOI":"10.1038\/nrdp.2016.94","volume":"3","author":"AMH Korver","year":"2017","unstructured":"Korver, A. M. H. et al. Congenital hearing loss. Nat. Rev. Dis. Primers 3(1), 16094 (2017).","journal-title":"Nat. Rev. Dis. Primers"},{"issue":"12","key":"83493_CR6","doi-asserted-by":"publisher","first-page":"1600","DOI":"10.1038\/gim.2018.48","volume":"20","author":"Q Guan","year":"2018","unstructured":"Guan, Q. et al. AUDIOME: a tiered exome sequencing-based comprehensive gene panel for the diagnosis of heterogeneous nonsyndromic sensorineural hearing loss. Genet. Med. 20(12), 1600\u20131608 (2018).","journal-title":"Genet. Med."},{"issue":"5","key":"83493_CR7","doi-asserted-by":"publisher","first-page":"536","DOI":"10.1038\/gim.2017.143","volume":"20","author":"NAM Almontashiri","year":"2018","unstructured":"Almontashiri, N. A. M. et al. Recurrent variants in OTOF are significant contributors to prelingual nonsydromic hearing loss in Saudi patients. Genet. Med. 20(5), 536\u2013544 (2018).","journal-title":"Genet. Med."},{"issue":"4","key":"83493_CR8","doi-asserted-by":"publisher","first-page":"441","DOI":"10.1007\/s00439-016-1648-8","volume":"135","author":"CM Sloan-Heggen","year":"2016","unstructured":"Sloan-Heggen, C. M. et al. Comprehensive genetic testing in the clinical evaluation of 1119 patients with hearing loss. Hum. Genet. 135(4), 441\u2013450 (2016).","journal-title":"Hum. Genet."},{"issue":"4","key":"83493_CR9","doi-asserted-by":"publisher","first-page":"364","DOI":"10.1038\/gim.2015.89","volume":"18","author":"G Bademci","year":"2016","unstructured":"Bademci, G. et al. Comprehensive analysis via exome sequencing uncovers genetic etiology in autosomal recessive nonsyndromic deafness in a large multiethnic cohort. Genet. Med. 18(4), 364\u2013371 (2016).","journal-title":"Genet. Med."},{"issue":"3","key":"83493_CR10","doi-asserted-by":"publisher","first-page":"308","DOI":"10.1038\/ejhg.2016.182","volume":"25","author":"C Zazo Seco","year":"2017","unstructured":"Zazo Seco, C. et al. The diagnostic yield of whole-exome sequencing targeting a gene panel for hearing impairment in The Netherlands. Eur. J. Hum. Genet. 25(3), 308\u2013314 (2017).","journal-title":"Eur. J. Hum. Genet."},{"issue":"1","key":"83493_CR11","doi-asserted-by":"publisher","first-page":"16783","DOI":"10.1038\/s41598-017-16846-9","volume":"7","author":"D Baux","year":"2017","unstructured":"Baux, D. et al. Combined genetic approaches yield a 48% diagnostic rate in a large cohort of French hearing-impaired patients. Sci. Rep. 7(1), 16783 (2017).","journal-title":"Sci. Rep."},{"key":"83493_CR12","doi-asserted-by":"publisher","first-page":"32","DOI":"10.1111\/cge.13754","volume":"98","author":"BS Budde","year":"2020","unstructured":"Budde, B. S. et al. Comprehensive molecular analysis of 61 Egyptian families with hereditary nonsyndromic hearing loss. Clin. Genet. 98, 32\u201342 (2020).","journal-title":"Clin. Genet."},{"issue":"11","key":"83493_CR13","doi-asserted-by":"publisher","first-page":"2442","DOI":"10.1038\/s41436-019-0535-9","volume":"21","author":"J Shen","year":"2019","unstructured":"Shen, J. et al. Consensus interpretation of the p.Met34Thr and p.Val37Ile variants in GJB2 by the ClinGen Hearing Loss Expert Panel. Genet. Med. 21(11), 2442\u20132452 (2019).","journal-title":"Genet. Med."},{"issue":"7","key":"83493_CR14","doi-asserted-by":"publisher","first-page":"565","DOI":"10.1038\/gim.2013.73","volume":"15","author":"RC Green","year":"2013","unstructured":"Green, R. C. et al. ACMG recommendations for reporting of incidental findings in clinical exome and genome sequencing. Genet. Med. 15(7), 565\u2013574 (2013).","journal-title":"Genet. Med."},{"key":"83493_CR15","doi-asserted-by":"crossref","unstructured":"Schiffmann, R. Fabry disease. Neurocutaneous Syndr. 2015:231\u2013248.","DOI":"10.1016\/B978-0-444-62702-5.00017-2"},{"issue":"18","key":"83493_CR16","doi-asserted-by":"publisher","first-page":"1870","DOI":"10.1001\/jama.2014.14601","volume":"312","author":"Y Yang","year":"2014","unstructured":"Yang, Y. et al. Molecular findings among patients referred for clinical whole-exome sequencing. JAMA 312(18), 1870\u20131879 (2014).","journal-title":"JAMA"},{"issue":"3","key":"83493_CR17","doi-asserted-by":"publisher","first-page":"328","DOI":"10.1016\/j.ajhg.2018.07.009","volume":"103","author":"MLB Schwartz","year":"2018","unstructured":"Schwartz, M. L. B. et al. A model for genome-first care: Returning secondary genomic findings to participants and their healthcare providers in a large research cohort. Am. J. Hum. Genet. 103(3), 328\u2013337 (2018).","journal-title":"Am. J. Hum. Genet."},{"issue":"6319","key":"83493_CR18","doi-asserted-by":"publisher","first-page":"aaf6814","DOI":"10.1126\/science.aaf6814","volume":"354","author":"FE Dewey","year":"2016","unstructured":"Dewey, F. E. et al. Distribution and clinical impact of functional variants in 50,726 whole-exome sequences from the DiscovEHR study. Science 354(6319), aaf6814 (2016).","journal-title":"Science"},{"issue":"2","key":"83493_CR19","doi-asserted-by":"publisher","first-page":"325","DOI":"10.1038\/s41431-018-0284-2","volume":"27","author":"L Haer-Wigman","year":"2019","unstructured":"Haer-Wigman, L. et al. 1 in 38 individuals at risk of a dominant medically actionable disease. Eur. J. Hum. Genet. 27(2), 325\u2013330 (2019).","journal-title":"Eur. J. Hum. Genet."},{"issue":"Suppl 1","key":"83493_CR20","doi-asserted-by":"publisher","first-page":"61S","DOI":"10.1177\/0003489415575060","volume":"124","author":"K Tsukada","year":"2015","unstructured":"Tsukada, K., Nishio, S. Y., Hattori, M. & Usami, S. Ethnic-specific spectrum of GJB2 and SLC26A4 mutations: Their origin and a literature review. Ann. Otol. Rhinol. Laryngol. 124(Suppl 1), 61S-76S (2015).","journal-title":"Ann. Otol. Rhinol. Laryngol."},{"issue":"10","key":"83493_CR21","doi-asserted-by":"publisher","first-page":"2231","DOI":"10.1038\/s41436-019-0481-6","volume":"21","author":"Q Wang","year":"2019","unstructured":"Wang, Q. et al. Nationwide population genetic screening improves outcomes of newborn screening for hearing loss in China. Genet. Med. 21(10), 2231\u20132238 (2019).","journal-title":"Genet. Med."},{"issue":"1","key":"83493_CR22","doi-asserted-by":"publisher","first-page":"52","DOI":"10.1089\/gtmb.2014.0241","volume":"19","author":"J Qing","year":"2015","unstructured":"Qing, J. et al. Prevalence of mutations in GJB2, SLC26A4, and mtDNA in children with severe or profound sensorineural hearing loss in southwestern China. Genet. Test Mol. Biomark. 19(1), 52\u201358 (2015).","journal-title":"Genet. Test Mol. Biomark."},{"key":"83493_CR23","doi-asserted-by":"publisher","first-page":"26","DOI":"10.1186\/1479-5876-7-26","volume":"7","author":"P Dai","year":"2009","unstructured":"Dai, P. et al. GJB2 mutation spectrum in 2,063 Chinese patients with nonsyndromic hearing impairment. J. Transl. Med. 7, 26 (2009).","journal-title":"J. Transl. Med."},{"issue":"3","key":"83493_CR24","doi-asserted-by":"publisher","first-page":"245","DOI":"10.1111\/j.1399-0004.2007.00862.x","volume":"72","author":"QJ Wang","year":"2007","unstructured":"Wang, Q. J. et al. A distinct spectrum of SLC26A4 mutations in patients with enlarged vestibular aqueduct in China. Clin. Genet. 72(3), 245\u2013254 (2007).","journal-title":"Clin. Genet."},{"key":"83493_CR25","doi-asserted-by":"publisher","first-page":"2614","DOI":"10.1038\/s41436-019-0563-5","volume":"21","author":"AE Shearer","year":"2019","unstructured":"Shearer, A. E., Shen, J., Amr, S., Morton, C. C. & Smith, R. J. Newborn Hearing Screening Working Group of the National Coordinating Center for the Regional Genetics Network. A proposal for comprehensive newborn hearing screening to improve identification of deaf and hard-of-hearing children. Genet. Med. 21, 2614\u20132630 (2019).","journal-title":"Genet. Med."},{"issue":"1","key":"83493_CR26","doi-asserted-by":"crossref","first-page":"7","DOI":"10.1101\/gr.9.1.7","volume":"9","author":"BJ Keats","year":"1999","unstructured":"Keats, B. J. & Berlin, C. I. Genomics and hearing impairment. Genome Res. 9(1), 7\u201316 (1999).","journal-title":"Genome Res."},{"issue":"11","key":"83493_CR27","doi-asserted-by":"publisher","first-page":"1593","DOI":"10.1002\/humu.23630","volume":"39","author":"AM Oza","year":"2018","unstructured":"Oza, A. M. et al. Expert specification of the ACMG\/AMP variant interpretation guidelines for genetic hearing loss. Hum. Mutat. 39(11), 1593\u20131613 (2018).","journal-title":"Hum. Mutat."},{"issue":"1","key":"83493_CR28","doi-asserted-by":"publisher","first-page":"21","DOI":"10.1056\/NEJMoa1516767","volume":"376","author":"JE Posey","year":"2017","unstructured":"Posey, J. E. et al. Resolution of disease phenotypes resulting from multilocus genomic variation. N. Engl. J. Med. 376(1), 21\u201331 (2017).","journal-title":"N. Engl. J. Med."},{"issue":"5","key":"83493_CR29","doi-asserted-by":"publisher","first-page":"37","DOI":"10.1186\/gm554","volume":"6","author":"AE Shearer","year":"2014","unstructured":"Shearer, A. E. et al. Copy number variants are a common cause of non-syndromic hearing loss. Genome Med. 6(5), 37 (2014).","journal-title":"Genome Med."},{"issue":"1","key":"83493_CR30","doi-asserted-by":"publisher","first-page":"30","DOI":"10.1186\/s13039-017-0333-5","volume":"10","author":"R Yao","year":"2017","unstructured":"Yao, R. et al. Evaluation of three read-depth based CNV detection tools using whole-exome sequencing data. Mol. Cytogenet. 10(1), 30 (2017).","journal-title":"Mol. Cytogenet."},{"issue":"4","key":"83493_CR31","doi-asserted-by":"publisher","first-page":"521","DOI":"10.1007\/s00439-020-02118-6","volume":"139","author":"L Guo","year":"2020","unstructured":"Guo, L. et al. Concurrent hearing and genetic screening in a general newborn population. Hum. Genet. 139(4), 521\u2013530 (2020).","journal-title":"Hum. Genet."},{"issue":"5","key":"83493_CR32","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1093\/gigascience\/gix024","volume":"6","author":"J Huang","year":"2017","unstructured":"Huang, J. et al. A reference human genome dataset of the BGISEQ-500 sequencer. Gigascience 6(5), 1\u20139 (2017).","journal-title":"Gigascience"},{"issue":"5","key":"83493_CR33","doi-asserted-by":"publisher","first-page":"491","DOI":"10.1038\/ng.806","volume":"43","author":"MA DePristo","year":"2011","unstructured":"DePristo, M. A. et al. A framework for variation discovery and genotyping using next-generation DNA sequencing data. Nat. Genet. 43(5), 491\u2013498 (2011).","journal-title":"Nat. Genet."},{"key":"83493_CR34","doi-asserted-by":"publisher","first-page":"2409","DOI":"10.1038\/s41436-019-0553-7","volume":"21","author":"MT DiStefano","year":"2019","unstructured":"DiStefano, M. T. et al. ClinGen expert clinical validity curation of 164 hearing loss gene\u2013disease pairs. Genet. Med. 21, 2409 (2019).","journal-title":"Genet. Med."},{"issue":"Database issue","key":"83493_CR35","doi-asserted-by":"publisher","first-page":"D670","DOI":"10.1093\/nar\/gku1177","volume":"43","author":"KR Rosenbloom","year":"2015","unstructured":"Rosenbloom, K. R. et al. The UCSC Genome Browser database: 2015 update. Nucleic Acids Res. 43(Database issue), D670\u2013D681 (2015).","journal-title":"Nucleic Acids Res."},{"key":"83493_CR36","doi-asserted-by":"publisher","first-page":"311","DOI":"10.1007\/978-1-61779-507-7_15","volume":"838","author":"S Ceulemans","year":"2012","unstructured":"Ceulemans, S., van der Ven, K. & Del-Favero, J. Targeted screening and validation of copy number variations. Methods Mol. Biol. 838, 311\u2013328 (2012).","journal-title":"Methods Mol. Biol."}],"container-title":["Scientific Reports"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.nature.com\/articles\/s41598-021-83493-6.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.nature.com\/articles\/s41598-021-83493-6","content-type":"text\/html","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.nature.com\/articles\/s41598-021-83493-6.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2022,12,2]],"date-time":"2022-12-02T23:02:01Z","timestamp":1670022121000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.nature.com\/articles\/s41598-021-83493-6"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2021,2,17]]},"references-count":36,"journal-issue":{"issue":"1","published-online":{"date-parts":[[2021,12]]}},"alternative-id":["83493"],"URL":"https:\/\/doi.org\/10.1038\/s41598-021-83493-6","relation":{"has-preprint":[{"id-type":"doi","id":"10.1101\/2020.09.20.20197145","asserted-by":"object"}]},"ISSN":["2045-2322"],"issn-type":[{"value":"2045-2322","type":"electronic"}],"subject":[],"published":{"date-parts":[[2021,2,17]]},"assertion":[{"value":"12 October 2020","order":1,"name":"received","label":"Received","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"25 January 2021","order":2,"name":"accepted","label":"Accepted","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"17 February 2021","order":3,"name":"first_online","label":"First Online","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"J.X., L.C., H.L., J.P., N.S., and Z.P. were employed at BGI Genomics at the time of submission. No other conflicts relevant to this study should be reported.","order":1,"name":"Ethics","group":{"name":"EthicsHeading","label":"Competing interests"}}],"article-number":"4036"}}