{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,1]],"date-time":"2026-07-01T10:50:53Z","timestamp":1782903053346,"version":"3.54.5"},"reference-count":28,"publisher":"Wiley","license":[{"start":{"date-parts":[[2026,6,30]],"date-time":"2026-06-30T00:00:00Z","timestamp":1782777600000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc-nd\/4.0\/"},{"start":{"date-parts":[[2026,6,30]],"date-time":"2026-06-30T00:00:00Z","timestamp":1782777600000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/doi.wiley.com\/10.1002\/tdm_license_1.1"}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Int J Lab Hematology"],"abstract":"ABSTRACT<\/jats:title>\n \n Background<\/jats:title>\n \n Hemophilia A, an X\u2010linked bleeding disorder caused by pathogenic variants in the\n F8<\/jats:italic>\n gene, requires precise genetic diagnosis for optimal management. Conventional stepwise sequence and copy number variation (CNV) analyses are time\u2010consuming and may leave some cases unresolved.\n <\/jats:p>\n <\/jats:sec>\n \n Objectives<\/jats:title>\n To evaluate the effectiveness of combining next\u2010generation sequencing (NGS)\u2010based sequence and CNV analysis in a single step for resolving previously undiagnosed Hemophilia A cases and identifying differential diagnoses.<\/jats:p>\n <\/jats:sec>\n \n Methods<\/jats:title>\n \n A cohort of 578 male patients with Factor VIII deficiency was assessed. Initial analyses included intron 22\/intron 1 inversion testing and\/or\n F8<\/jats:italic>\n sequencing. Twenty\u2010seven unresolved cases underwent a targeted 57\u2010gene coagulopathy panel using NGS with CNV analysis; CNVs were confirmed by multiplex ligation\u2010dependent probe amplification (MLPA).\n <\/jats:p>\n <\/jats:sec>\n \n Results<\/jats:title>\n \n The integrated NGS approach resolved 19 of 27 unresolved cases, increasing the overall diagnostic yield from 95.3% to 98.6%. Ten CNVs in\n F8<\/jats:italic>\n (8 deletions, 2 duplications) were identified and confirmed by MLPA. Variants in\n VWF<\/jats:italic>\n and\n LMAN1<\/jats:italic>\n genes led to diagnoses of von Willebrand disease (\n n<\/jats:italic>\n \u2009=\u20096) and combined Factor V\/VIII deficiency (\n n<\/jats:italic>\n \u2009=\u20091). One\n F8<\/jats:italic>\n intronic variant was detected, while 8 cases remained unresolved by the NGS\u2010based approach.\n <\/jats:p>\n <\/jats:sec>\n \n Conclusions<\/jats:title>\n Integrating NGS\u2010based CNV analysis into genetic diagnostic workflows enables simultaneous detection of sequence variants and CNVs, improves diagnostic yield, and facilitates identification of related bleeding disorders. This streamlined approach supports adoption as a first\u2010line test for mild\/moderate Hemophilia A and a secondary test for severe cases.<\/jats:p>\n <\/jats:sec>","DOI":"10.1111\/ijlh.70186","type":"journal-article","created":{"date-parts":[[2026,7,1]],"date-time":"2026-07-01T10:35:27Z","timestamp":1782902127000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Advanced Molecular Analysis in Hemophilia A in a Single Step: Next Generation Sequencing (\n NGS<\/scp>\n ) and Copy Number Variation (\n CNV<\/scp>\n ) Analysis"],"prefix":"10.1111","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-0582-8881","authenticated-orcid":false,"given":"Enise","family":"Avci Durmusalioglu","sequence":"first","affiliation":[{"name":"Department of Pediatric Genetics Ege University Faculty of Medicine Izmir Turkey"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0657-2408","authenticated-orcid":false,"given":"Esra","family":"Isik","sequence":"additional","affiliation":[{"name":"Department of Pediatric Genetics Ege University Faculty of Medicine Izmir Turkey"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Basak","family":"Durmus Ozen","sequence":"additional","affiliation":[{"name":"Department of Pediatric Genetics Ege University Faculty of Medicine Izmir Turkey"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mehmet Mert","family":"Topaloglu","sequence":"additional","affiliation":[{"name":"Department of Pediatric Genetics Ege University Faculty of Medicine Izmir Turkey"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nihal","family":"Karadas","sequence":"additional","affiliation":[{"name":"Department of Pediatric Hematology Ege University Faculty of Medicine Izmir Turkey"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4910-2142","authenticated-orcid":false,"given":"Kaan","family":"Kavakli","sequence":"additional","affiliation":[{"name":"Department of Pediatric Hematology Ege University Faculty of Medicine Izmir Turkey"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Melike","family":"Sezgin Evim","sequence":"additional","affiliation":[{"name":"Department of Pediatric Hematology Uludag University Faculty of Medicine Bursa Turkey"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tolga","family":"Aslan","sequence":"additional","affiliation":[{"name":"Genomize Bilisim ve Biyoteknoloji Anonim Sirketi Istanbul Turkey"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Er\u015fen","family":"Kavak","sequence":"additional","affiliation":[{"name":"Genomize Bilisim ve Biyoteknoloji Anonim Sirketi Istanbul Turkey"},{"name":"Department of Molecular Biology and Genetics, Faculty of Arts and Sciences Bogazici University Istanbul Turkey"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Adalet Meral","family":"Gunes","sequence":"additional","affiliation":[{"name":"Department of Pediatric Hematology Uludag University Faculty of Medicine Bursa Turkey"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2513-7188","authenticated-orcid":false,"given":"Funda","family":"Tayfun Kupesiz","sequence":"additional","affiliation":[{"name":"Department of Pediatric Hematology Akdeniz University Faculty of Medicine Antalya Turkey"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alphan","family":"Kupesiz","sequence":"additional","affiliation":[{"name":"Department of Pediatric Hematology Akdeniz University Faculty of Medicine Antalya Turkey"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Fahri","family":"Sahin","sequence":"additional","affiliation":[{"name":"Department of Hematology Ege University Faculty of Medicine Izmir Turkey"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tahir","family":"Atik","sequence":"additional","affiliation":[{"name":"Department of Pediatric Genetics Ege University Faculty of Medicine Izmir Turkey"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2026,6,30]]},"reference":[{"key":"e_1_2_11_2_1","doi-asserted-by":"publisher","DOI":"10.1111\/hae.14046"},{"key":"e_1_2_11_3_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41572\u2010021\u201000278\u2010x"},{"key":"e_1_2_11_4_1","doi-asserted-by":"publisher","DOI":"10.1111\/jth.15805"},{"key":"e_1_2_11_5_1","volume-title":"GeneReviews","author":"Konkle B. 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