The spectrum of FVIII gene variants detected by next generation sequencing in 236 Chinese non-inversion hemophilia A pedigrees

Chen, Juanjuan; Li, Qiang; Lin, Shiu-Shiung; Li, Fenxia; Huang, Lv‐Yin; Jin, Wangjie; Yang, Xu; Li, Yihong; Li, Kun; Xiong, Yufeng; Fan, Daiming; Luo, Dixian; Li, Liyan

doi:10.1016/j.thromres.2021.02.027

Cited by 5 publications

(4 citation statements)

References 23 publications

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“…In addition, no disease-causing variant could be identified in the respective genes in 2% of our patients with hemophilia A and 1% of patients with hemophilia B, which is comparable with the CHAMP data and with some reports [ [24] , [25] , [26] ], but low in comparison with other studies, that failed to identify a disease-causing variant in up to 11% of the patients. [ [27] , [28] , [29] , [30] , [31] ] Of note, recent evolutions in genetic testing have improved variant detection and may explain the differences between various studies.…”

Section: Discussionmentioning

confidence: 99%

F8/F9 variants in the population-based PedNet Registry cohort compared with locus-specific genetic databases of the European Association for Haemophilia and Allied Disorders and the Centers for Disease Control and Prevention Hemophilia A or Hemophilia B Mutation Project

Labarque¹,

Mancuso²,

Kartal-Kaess³

et al. 2023

Research and Practice in Thrombosis and Haemostasis

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Section: Discussionmentioning

confidence: 99%

F8/F9 variants in the population-based PedNet Registry cohort compared with locus-specific genetic databases of the European Association for Haemophilia and Allied Disorders and the Centers for Disease Control and Prevention Hemophilia A or Hemophilia B Mutation Project

Labarque¹,

Mancuso²,

Kartal-Kaess³

et al. 2023

Research and Practice in Thrombosis and Haemostasis

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“…Prenatal testing is crucial for the mothers of affected patients in order to prevent the recurrence of variants in subsequent pregnancies. To date, numerous F8 and F9 variants have been reported in different populations, including Chinese cohorts ( https://dbs.eahad.org/ ) ( Xue et al, 2010 ; Guo et al, 2018 ; Luna-Záizar et al, 2018 ; Chen et al, 2021 ). This study aimed to investigate the clinical and genetic features of hemophilia in southern China by recruiting patients with HA and HB and conducting clinical as well as genetic analyses.…”

Section: Introductionmentioning

confidence: 99%

Variant spectrum of F8 and F9 in hemophilia patients from southern China and 26 novel variants

Li,

He,

Chen

et al. 2023

Front. Genet.

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Hemophilia, an X-linked recessive disorder, is characterized by spontaneous or trauma-induced prolonged bleeding. It is classified as hemophilia A when caused by variants in the F8 gene, and hemophilia B when caused by F9 variants. Few studies have described hemophilia variants in the Chinese population. This study aimed to investigate the clinical and genetic profiles of 193 hemophilia patients from southern China. Utilizing Sanger sequencing, multiplex ligation-dependent probe amplification, gap detection, long-range PCR, and multiplex PCR, we identified both F8 and F9 gene variants. Pregnant women with a history of hemophilia A offspring underwent amniocentesis or villus sampling for the variant detection. Variants in F8 and F9 were pinpointed in 183 patients, with 26 being novel discoveries. Notably, genetic testing was absent in the initial evaluation of 133 out of 161 patients, leading to a protracted average definitive diagnosis timeline of 2 years. Remarkably, two hemophilia A cases with anticipated severe phenotypes due to protein-truncating variants presented with only moderate or mild clinical manifestations. Among the 40 fetuses tested, 34 were males, with 17 exhibiting hemizygous variants in the F8 gene. Our results contribute to the broader understanding of F8 and F9 variant spectrum and highlight the underuse of genetic analyses in southern China.

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“… 20 21 In recent years, next-generation sequencing (NGS) has been employed for high-throughput genetic screening of SNVs and indels of F8 including the deep intronic regions. 22 23 24 When PCR failure and NGS suggest the absence of one or more exons in male patients, multiplex ligation-dependent probe amplification (MLPA) allows confirmation of deletions. MLPA can also define the carrier status in female relatives for large gene deletions, being masked by the amplification of the normal allele using PCR, as well as duplications that are unidentifiable by standard PCR.…”

Section: Introductionmentioning

confidence: 99%

“…For patients negative for Inv22, different methods were then applied for Inv1, SNVs/indels, and large deletions/duplications. 20 22 26 27 28 While overall cost-effective, the whole diagnosis process can take a long time for patients negative for inversions. To overcome the limit of short-read length in detecting highly homologous regions, Johnsen et al developed a NGS-based approach that captured Inv22 and Inv1 from Ksp221 digested/ligated DNA, and other variants from untreated genomic DNA simultaneously.…”

Section: Introductionmentioning

confidence: 99%

Comprehensive Analysis of Hemophilia A (CAHEA): Towards Full Characterization of the F8 Gene Variants by Long-Read Sequencing

Liu

et al. 2023

Thromb Haemost

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Hemophilia A (HA) is the most frequently occurring X-linked bleeding disorder caused by heterogeneous variants in the F8 gene, one of the largest genes known. Conventional molecular analysis of F8 requires a combination of assays, usually including long range (LR)-PCR or inverse PCR for inversions, Sanger sequencing or next-generation sequencing for single-nucleotide variants (SNVs) and indels, and multiplex ligation-dependent probe amplification for large deletions or duplications. This study aimed to develop a LR-PCR and long-reads sequencing (LRS)-based assay termed comprehensive analysis of hemophilia A (CAHEA) for full characterization of F8 variants. The performance of CAHEA was evaluated in 272 samples from 131 HA pedigrees with a wide spectrum of F8 variants by comparing to conventional molecular assays. CAHEA identified F8 variants in all the 131 pedigrees, including 35 intron 22-related gene rearrangements, three intron 1 inversion (Inv1), 85 SNVs and indels, one large insertion and seven large deletions. Compared to the conventional methods combined altogether, CAHEA assay demonstrated 100% sensitivity and specificity for identifying various type of F8 variants and had the advantages of directly determining the break regions/points of large inversions, insertions and deletions, which enabled analyzing the mechanisms of recombination at the junction sites and pathogenicity of the variants. CAHEA represents a comprehensive assay towards full characterization of F8 variants including intron 22 and intron 1 inversions, SNVs/indels, large insertions and deletions, greatly improving the genetic screening and diagnosis for HA.

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The spectrum of FVIII gene variants detected by next generation sequencing in 236 Chinese non-inversion hemophilia A pedigrees

Cited by 5 publications

References 23 publications

F8/F9 variants in the population-based PedNet Registry cohort compared with locus-specific genetic databases of the European Association for Haemophilia and Allied Disorders and the Centers for Disease Control and Prevention Hemophilia A or Hemophilia B Mutation Project

F8/F9 variants in the population-based PedNet Registry cohort compared with locus-specific genetic databases of the European Association for Haemophilia and Allied Disorders and the Centers for Disease Control and Prevention Hemophilia A or Hemophilia B Mutation Project

Variant spectrum of F8 and F9 in hemophilia patients from southern China and 26 novel variants

Comprehensive Analysis of Hemophilia A (CAHEA): Towards Full Characterization of the F8 Gene Variants by Long-Read Sequencing

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