Identification of 217 unreported mutations in the F8 gene in a group of 1,410 unselected Italian patients with hemophilia A

Abstract

To provide a National database, 1,410 unrelated hemophilia A (HA) patients were investigated using screening methods denaturing high-performance liquid chromatography (DHPLC), conformational-sensitive gel electrophoresis (CSGE)] and/or direct sequencing. F8 gene mutations were identified in 877 (81%), 146 (82%), and 133 (89%) families with severe, moderate, or mild HA, respectively. Among the 382 different mutations detected, 217 (57%) have not previously been reported in the F8 Haemophilia A Mutation, Structure, Test and Resource Site (HAMSTeRS) database. Mutations leading to a null allele accounted for 82, 15%, and less than 1% of severe, moderate, or mild HA, respectively. A missense mutation was identified in 16%, 68%, and 81% of severe, moderate, or mild HA, respectively. They included 105 missense mutations (48%), 41 small deletions (19%), 25 splice site mutations (12%), 24 nonsense mutations (11%), 18 insertions (8%), three large deletions (1%), and one deletion plus insertion. Unreported mutations were distributed throughout the F8 gene, as they affected all F8 exons but exon 20. We report a wide spectrum of mutations collected in a large National database. The type of mutation was a strong predictor of the clinical phenotype. This database is expected to considerably improve the genetic counseling and medical care of HA families in Italy.

Genotyping of Intron Inversions and Point Mutations in Exon 14 of the FVIII Gene in Iranian Azeri Turkish Families with Hemophilia A

“…Exon 14, with a length of 3 kb, accounts for approximately 43 % of the coding region of the F8, and many different mutations have been reported in this portion of the gene. Our findings explain the occurrence of a few mutations in exon 14, and our study has a little discrepancy with a previous report [28]. Identification of carriers in all families with HA is a very important issue and would improve the genetic counseling and quality of life of these families.…”

“…In previous reports, gross exon deletions may be comprised of almost 4 % of mutations in patients with severe HA [28]. In the present study, all patients were normal for gross deletions.…”

Factor VIII genetic mutations and protein alterations in hemophilia A: A review

“…We have also found the change of arginine 1781 to histidine in 3 unrelated patients in A3 domain (Faridi et al, 2011). Santacroce et al (2008) were able to detected 384 and 67 (Feng et al, 2010) different mutations and these were mostly effected in the A domains (Table 2).…”

Chapter 1 Profiling of Mutations in the F8 and F9, Causative Genes of Hemophilia A and Hemophilia B"

“…The strategy for direct sequencing analysis is largely similar to that for HA, but is simpler and less costly because the F9 gene is smaller and is encoded by a smaller number of exons. In addition, as in HA, mutation scanning by CSGE can also be applied for direct sequencing analyses, but the detection sensitivity of CSGE needs to be validated in each laboratory prior to clinical implementation (Santacroce et al, 2008). Large deletion mutations, which can be detected by MLPA analyses, should be suspected when 1 or more reactions to amplify a genomic segment fail.…”