Skip to content
DraconiticResearch
Search Sources API
203,234 studies31 sourcesUpdated 17 min ago
Peer-reviewedCC BY-NC-ND 4.0International Journal of Laboratory Hematology

Advanced Molecular Analysis in Hemophilia A in a Single Step: Next Generation Sequencing (NGS) and Copy Number Variation (CNV) Analysis

  • Enise Avci Durmusalioglu1
  • Esra Isik1
  • Basak Durmus Ozen1
  • Mehmet Mert Topaloglu1
  • Nihal Karadas2
  • Kaan Kavakli2
  • Melike Sezgin Evim3
  • Tolga Aslan4
  • Erşen Kavak4,5
  • Adalet Meral Gunes3
  • Funda Tayfun Kupesiz6
  • Alphan Kupesiz6
2 more authors
  • Fahri Sahin7
  • Tahir Atik1
7 affiliations
  1. 1Department of Pediatric Genetics Ege University Faculty of Medicine Izmir Turkey
  2. 2Department of Pediatric Hematology Ege University Faculty of Medicine Izmir Turkey
  3. 3Department of Pediatric Hematology Uludag University Faculty of Medicine Bursa Turkey
  4. 4Genomize Bilisim ve Biyoteknoloji Anonim Sirketi Istanbul Turkey
  5. 5Department of Molecular Biology and Genetics, Faculty of Arts and Sciences Bogazici University Istanbul Turkey
  6. 6Department of Pediatric Hematology Akdeniz University Faculty of Medicine Antalya Turkey
  7. 7Department of Hematology Ege University Faculty of Medicine Izmir Turkey
Read full textDOICite

Abstract

Background: Hemophilia A, an X‐linked bleeding disorder caused by pathogenic variants in the F8 gene, requires precise genetic diagnosis for optimal management. Conventional stepwise sequence and copy number variation (CNV) analyses are time‐consuming and may leave some cases unresolved.

Objectives: To evaluate the effectiveness of combining next‐generation sequencing (NGS)‐based sequence and CNV analysis in a single step for resolving previously undiagnosed Hemophilia A cases and identifying differential diagnoses.

Methods: A cohort of 578 male patients with Factor VIII deficiency was assessed. Initial analyses included intron 22/intron 1 inversion testing and/or F8 sequencing. Twenty‐seven unresolved cases underwent a targeted 57‐gene coagulopathy panel using NGS with CNV analysis; CNVs were confirmed by multiplex ligation‐dependent probe amplification (MLPA).

Results: The integrated NGS approach resolved 19 of 27 unresolved cases, increasing the overall diagnostic yield from 95.3% to 98.6%. Ten CNVs in F8 (8 deletions, 2 duplications) were identified and confirmed by MLPA. Variants in VWF and LMAN1 genes led to diagnoses of von Willebrand disease (n = 6) and combined Factor V/VIII deficiency (n = 1). One F8 intronic variant was detected, while 8 cases remained unresolved by the NGS‐based approach.

Conclusions: Integrating NGS‐based 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‐line test for mild/moderate Hemophilia A and a secondary test for severe cases.

Related

  1. Molecular overlap between primary immune thrombocytopenia and inherited platelet disorders: A pediatric case report with a novel heterozygous ITGB3 variant identified by next-generation sequencing
    Félix David Duarte Rodríguez, Óscar Eduardo González Figueredo, Alejandra Calderón Gasca
    Therapeutic Advances in HematologyPeer-reviewedCC BY-NC 4.0Full textGeneral medicinePMCEurope PMC
  2. Longitudinal TP53 dynamics in repeated tissue-based next-generation sequencing in non-small cell lung cancer: a real-world study with molecular lineage adjudication and sequential credibility verification.
    Dan Zhao, Deyang Xie, Kun Li +3
    Journal of thoracic diseasePeer-reviewedCC BY-NC-NDFull textGeneral medicineEurope PMC
  3. Quantifying the diagnostic time gap in precision oncology: A retrospective analysis of potential time savings from same-day cartridge-based molecular testing versus next-generation sequencing in cytology specimens
    Kevin Tran, Eveline Chen, W. Fraser Symmans +1
    PLOS OnePeer-reviewedCC BY 4.0Full textOncologyPLOS
  4. Diagnostic performance of next-generation sequencing in thyroid nodules and molecular correlates of tumor aggressiveness in papillary thyroid carcinoma
    Songze Zhang, Nan Zhao, Lan Qin +1
    Frontiers in EndocrinologyPeer-reviewedCC BY 4.0Full textEndocrinology & metabolismCrossrefEurope PMCPMC
  5. Rare copy number variation in Post-traumatic stress disorder using whole genome sequencing and biobank data
    S. Kramer, S. Farajzadeh Valilou, M. Singh +12
    medRxivPreprintAll rights reservedFull textHuman geneticsmedRxiv

Cite

BibTeXRIS
BibTeX
@article{durmusalioglu2026advanced,
  title = {{Advanced Molecular Analysis in Hemophilia A in a Single Step: Next Generation Sequencing (NGS) and Copy Number Variation (CNV) Analysis}},
  author = {Enise Avci Durmusalioglu and Esra Isik and Basak Durmus Ozen and Mehmet Mert Topaloglu and Nihal Karadas and Kaan Kavakli and Melike Sezgin Evim and Tolga Aslan and Erşen Kavak and Adalet Meral Gunes and Funda Tayfun Kupesiz and Alphan Kupesiz and Fahri Sahin and Tahir Atik},
  journal = {International Journal of Laboratory Hematology},
  year = {2026},
  month = jun,
  doi = {10.1111/ijlh.70186},
  url = {https://doi.org/10.1111/ijlh.70186},
  note = {CC BY-NC-ND 4.0},
}
RIS
TY  - JOUR
TI  - Advanced Molecular Analysis in Hemophilia A in a Single Step: Next Generation Sequencing (NGS) and Copy Number Variation (CNV) Analysis
AU  - Enise Avci Durmusalioglu
AU  - Esra Isik
AU  - Basak Durmus Ozen
AU  - Mehmet Mert Topaloglu
AU  - Nihal Karadas
AU  - Kaan Kavakli
AU  - Melike Sezgin Evim
AU  - Tolga Aslan
AU  - Erşen Kavak
AU  - Adalet Meral Gunes
AU  - Funda Tayfun Kupesiz
AU  - Alphan Kupesiz
AU  - Fahri Sahin
AU  - Tahir Atik
PY  - 2026
DA  - 2026/06/30
JO  - International Journal of Laboratory Hematology
DO  - 10.1111/ijlh.70186
UR  - https://doi.org/10.1111/ijlh.70186
AB  - Background: Hemophilia A, an X‐linked bleeding disorder caused by pathogenic variants in the F8 gene, requires precise genetic diagnosis for optimal management. Conventional stepwise sequence and copy number variation (CNV) analyses are time‐consuming and may leave some cases unresolved.  Objectives: To evaluate the effectiveness of combining next‐generation sequencing (NGS)‐based sequence and CNV analysis in a single step for resolving previously undiagnosed Hemophilia A cases and identifying differential diagnoses.  Methods: A cohort of 578 male patients with Factor VIII deficiency was assessed. Initial analyses included intron 22/intron 1 inversion testing and/or F8 sequencing. Twenty‐seven unresolved cases underwent a targeted 57‐gene coagulopathy panel using NGS with CNV analysis; CNVs were confirmed by multiplex ligation‐dependent probe amplification (MLPA).  Results: The integrated NGS approach resolved 19 of 27 unresolved cases, increasing the overall diagnostic yield from 95.3% to 98.6%. Ten CNVs in F8 (8 deletions, 2 duplications) were identified and confirmed by MLPA. Variants in VWF and LMAN1 genes led to diagnoses of von Willebrand disease (n = 6) and combined Factor V/VIII deficiency (n = 1). One F8 intronic variant was detected, while 8 cases remained unresolved by the NGS‐based approach.  Conclusions: Integrating NGS‐based 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‐line test for mild/moderate Hemophilia A and a secondary test for severe cases.
SN  - 1751-5521
SN  - 1751-553X
ER  -