Identification of factor VIII gene mutations in patients with haemophilia A

Abstract

This study aimed to identify common FVIII gene mutations in Iraqi patients by molecular analysis and to investigate the correlations between mutations and disease severity. This study included 25 Iraqi patients with haemophilia A, 18 of whom were male haemophilia patients and 7 of whom were carrier mothers, from seven unrelated families. After determining the purity and concentration of the extracted DNA, selected areas of the FVIII gene, including exons 18, 22, 23, and 24 and intron 22, were amplified. Sequencing of all exons and intron 22 was conducted in all patients and controls. Sequencing analysis revealed that many mutations were distributed among the exons (18, 22, 23, and 24) and intron 22; most of the identified mutations were point mutations in exons, predominantly in exon 24. Exon 18 mutations were detected in one male and one mother as a carrier. Exon 22 mutations were detected in four patients, and exon 23 mutations were detected in two males and two carrier mothers. Exon 24 mutations were detected in 12 haemophilia patients and two carrier mothers. Additionally, seven inversion mutations were detected in haemophilia patients. The results also revealed positive correlations between disease severity and mutations in intron 22 and exon 24. We therefore concluded that exon 24 mutations are the most frequent type of mutation that occurs in haemophilia patients in Iraq, followed by intron 22 mutations, and that almost all severe haemophilia cases have mutations located in these two gene segments. Keywords: Camptothecin, Carotenoid, Cellulase enzyme, Extraction, HPLC