Identification of seven novel mutations of F8C by DHPLC.
Hemophilia A is an X-linked recessive disorder resulting from deficiency of Factor VIII (F8C), an important protein in blood coagulation. A large number of disease producing mutations have been reported in the F8C gene. However, a comprehensive analysis of mutations is difficult to conduct due to the large gene size, its many scattered exons, and the high frequency of de novo mutations. In this study, we performed analysis using PCR, Conformation Sensitive Gel Electrophoresis (CSGE), Denaturing High Performance Liquid Chromatography (DHPLC) and direct sequencing. We found seven novel mutations causing severe, moderate and mild Hemophilia A: IVS14-1G>A, G458V, T1695S, L1758P, Q2311P, 1441delT, 1269-1271insA. At least four variants detected by DHPLC (IVS14-1G>A, Q2311P,_R698W and D1241Q) were not detectable by CSGE.
Frusconi
Sabrina
S
Cytogenetic and Genetic Unit, "Careggi" Hospital, Florence, Italy. citogenbibl@ao-careggi.toscana.it
Passerini
Ilaria
I
Girolami
Francesca
F
Masieri
Maddalena
M
Linari
Silvia
S
Longo
Giovanni
G
Morfini
Massimo
M
Torricelli
Francesca
F
D000595:Amino Acid Sequence; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004247:DNA / Q000737:chemistry / Q000235:genetics; D004252:DNA Mutational Analysis; D005169:Factor VIII / Q000235:genetics*; D006467:Hemophilia A / Q000235:genetics* / Q000473:pathology; D006801:Humans; D008969:Molecular Sequence Data; D009154:Mutation; D017386:Sequence Homology, Amino Acid
D005169:Factor VIII; D004247:DNA