{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,5,20]],"date-time":"2026-05-20T03:29:52Z","timestamp":1779247792887,"version":"3.51.4"},"reference-count":23,"publisher":"Wiley","issue":"9","license":[{"start":{"date-parts":[[2020,7,9]],"date-time":"2020-07-09T00:00:00Z","timestamp":1594252800000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Grant of The Reproductive and Genetic Hospital of CITIC-Xiangya","award":["KYXM-201605"],"award-info":[{"award-number":["KYXM-201605"]}]},{"name":"Science and Technology Major Project of the Ministry of Science and Technology of Hunan Province, China","award":["2017SK1032"],"award-info":[{"award-number":["2017SK1032"]}]},{"DOI":"10.13039\/501100004761","name":"Natural Science Foundation of Hunan Province","doi-asserted-by":"crossref","award":["2019JJ50397"],"award-info":[{"award-number":["2019JJ50397"]}],"id":[{"id":"10.13039\/501100004761","id-type":"DOI","asserted-by":"crossref"}]}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Molec Gen &amp;amp; Gen Med"],"published-print":{"date-parts":[[2020,9]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:sec><jats:title>Background<\/jats:title><jats:p>Prezygotic de novo mutations may be inherited from parents with germline mosaicism and are often overlooked when the resulting phenotype affects only one child. We aimed to identify paternal germline mosaicism in an index family and provide a strategy to determine germline mosaicism.\u2018<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods<\/jats:title><jats:p>Whole\u2010exome sequencing was performed on an Alport syndrome\u2010affected child. Variants were validated using Sanger sequencing in the pedigree analysis. An apparent de novo mutation was tested by next\u2010generation sequencing (NGS) following chromosome microdissection of the mutant region (MicroSeq) to clarify its homologous chromosome source. Mosaic mutation in sperm samples was detected using targeted next\u2010generation sequencing (TNGS). Self\u2010prepared mosaic DNA samples of the 3% and 0.1% mutant fractions were used to evaluate the TNGS detection sensitivity.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>Two novel heterozygous variants, maternally inherited c.1322delT (p.Ile441Thrfs*17) and the de novo mutation c.2939T&gt;A (p.Leu980Ter), in the <jats:italic>COL4A3<\/jats:italic> gene were discovered in the propositus. MicroSeq identified c.2939T&gt;A in the paternal chromosome, which was <jats:italic>in trans<\/jats:italic> with c.1322delT. The frequency of c.2937A was 2.65% in the father's sperm sample. We also showed that a 500X depth coverage may detect a mosaic mutation with an allele frequency as low as 2%\u20133% using TNGS.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusion<\/jats:title><jats:p>MicroSeq is a valuable tool to identify the allele source of de novo mutations in a single patient. TNGS can be used to assess the mosaic ratios of known sites. We provided a systematic algorithm to detect germinal mosaicism in a single patient. This algorithm may have implications for genetic and reproductive counseling on germline mosaicism.<\/jats:p><\/jats:sec>","DOI":"10.1002\/mgg3.1394","type":"journal-article","created":{"date-parts":[[2020,7,9]],"date-time":"2020-07-09T10:00:56Z","timestamp":1594288856000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":11,"title":["Identification of paternal germline mosaicism by MicroSeq and targeted next\u2010generation sequencing"],"prefix":"10.1002","volume":"8","author":[{"given":"Congling","family":"Dai","sequence":"first","affiliation":[{"name":"Institute of Reproductive and Stem Cell Engineering School of Basic Medical Science Central South University Hunan China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Dehua","family":"Cheng","sequence":"additional","affiliation":[{"name":"Institute of Reproductive and Stem Cell Engineering School of Basic Medical Science Central South University Hunan China"},{"name":"Reproductive and Genetic Hospital ofCITIC\u2010Xiangya Hunan China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Weina","family":"Li","sequence":"additional","affiliation":[{"name":"Reproductive and Genetic Hospital ofCITIC\u2010Xiangya Hunan China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0111-8193","authenticated-orcid":false,"given":"Sicong","family":"Zeng","sequence":"additional","affiliation":[{"name":"Reproductive and Genetic Hospital ofCITIC\u2010Xiangya Hunan China"},{"name":"School of medicine Hunan Normal University Hunan China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Guangxiu","family":"Lu","sequence":"additional","affiliation":[{"name":"Institute of Reproductive and Stem Cell Engineering School of Basic Medical Science Central South University Hunan China"},{"name":"Reproductive and Genetic Hospital ofCITIC\u2010Xiangya Hunan China"},{"name":"School of medicine Hunan Normal University Hunan China"},{"name":"Hunan Guangxiu Hospital Hunan China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3391-5413","authenticated-orcid":false,"given":"Qianjun","family":"Zhang","sequence":"additional","affiliation":[{"name":"Institute of Reproductive and Stem Cell Engineering School of Basic Medical Science Central South University Hunan China"},{"name":"Reproductive and Genetic Hospital ofCITIC\u2010Xiangya Hunan China"},{"name":"Hunan Guangxiu Hospital Hunan China"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2020,7,9]]},"reference":[{"key":"e_1_2_9_2_1","doi-asserted-by":"publisher","DOI":"10.1111\/cge.12156"},{"key":"e_1_2_9_3_1","doi-asserted-by":"publisher","DOI":"10.1002\/ajmg.a.20568"},{"key":"e_1_2_9_4_1","doi-asserted-by":"publisher","DOI":"10.1136\/jmg.26.9.553"},{"key":"e_1_2_9_5_1","doi-asserted-by":"publisher","DOI":"10.1126\/science.2349482"},{"key":"e_1_2_9_6_1","doi-asserted-by":"publisher","DOI":"10.1002\/humu.22685"},{"key":"e_1_2_9_7_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.tig.2015.03.013"},{"key":"e_1_2_9_8_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.ajhg.2014.07.003"},{"key":"e_1_2_9_9_1","doi-asserted-by":"publisher","DOI":"10.1038\/ki.1985.63"},{"key":"e_1_2_9_10_1","doi-asserted-by":"publisher","DOI":"10.1038\/ejhg.2015.113"},{"key":"e_1_2_9_11_1","doi-asserted-by":"publisher","DOI":"10.1016\/0002\u20109343(81)90571\u20104"},{"key":"e_1_2_9_12_1","doi-asserted-by":"publisher","DOI":"10.1111\/cga.12279"},{"key":"e_1_2_9_13_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.ebiom.2016.11.007"},{"key":"e_1_2_9_14_1","doi-asserted-by":"publisher","DOI":"10.1186\/1471\u20102407\u201014\u201013"},{"key":"e_1_2_9_15_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.ejmg.2016.08.009"},{"key":"e_1_2_9_16_1","doi-asserted-by":"publisher","DOI":"10.1177\/0003489415575062"},{"key":"e_1_2_9_17_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.nmd.2014.04.002"},{"key":"e_1_2_9_18_1","doi-asserted-by":"publisher","DOI":"10.1038\/ng0994\u201077"},{"key":"e_1_2_9_19_1","doi-asserted-by":"publisher","DOI":"10.1002\/em.2850250606"},{"key":"e_1_2_9_20_1","doi-asserted-by":"publisher","DOI":"10.1007\/s40620\u2010018\u20100518\u2010y"},{"key":"e_1_2_9_21_1","doi-asserted-by":"publisher","DOI":"10.1136\/jmg.37.3.238"},{"key":"e_1_2_9_22_1","doi-asserted-by":"publisher","DOI":"10.1038\/gim.2015.30"},{"key":"e_1_2_9_23_1","doi-asserted-by":"publisher","DOI":"10.1007\/bf00286105"},{"key":"e_1_2_9_24_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1523\u20101755.2000.00358.x"}],"container-title":["Molecular Genetics &amp; 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