First report of molecular diagnosis of Tunisian hemophiliacs A: Identification of 8 novel causative mutations

Diagnostic Pathology

Emna Gouider²  Amel Ben Ammar Elggaaied³  Balkis Meddeb²  Christine Vinciguerra¹  Dorothé Pellechia¹  Kaouther Zahra²  Asma Jlizi³  Edvard Wigren⁴  Houssein Khodjet-el-khil³  Hejer Elmahmoudi³ 

[1] Biological hematology department EAM 4174, Hospices Civils de Lyon Université de Lyon, Lyon, France;Hemophilia Treatment Center, Aziza Othmana Hospital, Tunis, Tunisia;Laboratory of Genetics, Immunology and Human Pathologies, Tunis, Tunisia;Department of Medical Biochemistry and Biophysics, Karolinska Institutet, Stockholm, Sweden

关键词: Tunisia;    Molecular analysis;    Inhibitors;    Intron 1 inversion;    Intron 22 inversion;    Mutations;    Hemophilia A;   

Others  :  808014
DOI  :  10.1186/1746-1596-7-93

 received in 2012-06-20, accepted in 2012-07-17,  发布年份 2012

【 摘 要 】

Introduction

Hemophilia A is an X linked recessive hemorrhagic disorder caused by mutations in the F8 gene that lead to qualitative and/or quantitative deficiencies of coagulation factor VIII (FVIII). Molecular diagnosis of hemophilia A is challenging because of the high number of different causative mutations that are distributed throughout the large F8 gene. Molecular studies of these mutations are essential in order to reinforce our understanding of their pathogenic effect responsible for the disorder.

Aim

In this study we have performed molecular analysis of 28 Tunisian hemophilia A patients and analyzed the F8 mutation spectrum.

Methods

We screened the presence of intron 22 and intron 1 inversion in severe hemophilia A patients by southern blotting and polymerase chain reaction (PCR). Detection of point mutations was performed by dHPLC/sequencing of the coding F8 gene region. We predict the potential functional consequences of novel missense mutations with bioinformatics approaches and mapping of their spatial positions on the available FVIII 3D structure.

Results

We identified 23 different mutations in 28 Tunisian hemophilia A patients belonging to 22 unrelated families. The identified mutations included 5 intron 22 inversions, 7 insertions, 4 deletions and 7 substitutions. In total 18 point mutations were identified, of which 9 are located in exon 14, the most mutated exonic sequence in the F8 gene. Among the 23 mutations, 8 are novel and not deposited in the HAMSTeRS database nor described in recently published articles.

Conclusion

The mutation spectrum of Tunisian hemophilia A patients is heterogeneous with the presence of some characteristic features.

Virtual slides

The virtual slide(s) for this article can be found here:

http://www.diagnosticpathology.diagnomx.eu/vs/1693269827490715 webcite

【 授权许可】

   
2012 Elmahmoudi et al.; licensee BioMed Central Ltd.

【 图 表 】

Figure 1.

Figure 2.

Figure 3.

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