ملخص
Abstract
This study is to identify some mutations in exons 23, 24, 26 A+B, and 26 A of the factor VIII gene that causes hemophilia A among Libyan patients. Forty-three patients (40 males, 3 females), 10 carrier, and 10 control (8 males, 2 females) were enrolled in this study. The total number were 63 samples for each exon. Blood samples were collected from Libyan patients (Department of Paediatric Hematology, Tripoli Medical Center), and kept in freezer at -20 Cº. The blood samples were transported to DNA laboratory in the Genetic Engineering Department and Human Tissues Department Biotechnology Research Center, Twesha- Libya. Genomic DNA was extracted from whole blood and DNA was visualized by Agarose Gel Electrophoresis, and DNA concentration was measured by Spectrophotometer. DNA was amplified by polymerase chain reaction (PCR) technique. Mutations were identified by restriction enzymes Taq I and Xba I. Taq I was used to detect nonsense mutations in Exon 23, 2147 [ Arg Stop codon (Term)], Exon 24, 2209 [ Arg Stop codon (Term)], and Exon 26 A+B, 2307 [ Arg Stop codon (Term)]. Our result showed no mutations were detected by using restriction enzyme Taq I in Exons 23, codon 2147, Exon 24, codon 2209, and Exon 26A+B, codon 2307. But by using restriction enzyme Xba I, we found six insertion mutations in Exon 26A, 3919 (ins GA) 11.3 % of 53 patients, and two partial deletion mutations in Exon 23 (3.8 % of 53 patients) refer to area failed to be amplified by PCR.