Identification of mutations in 15 Hungarian families with hereditary protein C deficiency

• Published in: British journal of haematology 2000-10, Vol.111 (1), p.129-135
• Main Authors: DAVID, M, LOSONCZY, H, SAS, G, NAGY, A, KUTSCHER, G, MEYER, M
• Format: Article
• Language: English
• Subjects: Adult; Biological and medical sciences; Electrophoresis, Polyacrylamide Gel; Exons; Female; Frameshift Mutation; Gene Deletion; Hematologic and hematopoietic diseases; Hematology; Humans; Male; Medical sciences; Middle Aged; Mutation, Missense; Nucleic Acid Denaturation; Platelet diseases and coagulopathies; Polymerase Chain Reaction; Polymorphism, Single-Stranded Conformational; Protein C - genetics; Protein C Deficiency - genetics; Sequence Analysis, DNA; Venous Thrombosis - genetics
• ISSN: 0007-1048; 1365-2141
• DOI: 10.1046/j.1365-2141.2000.02324.x

Hereditary protein C deficiency represents about 2-9% of inherited thrombophilias. Our aim was to elucidate the molecular basis of protein C deficiency in 25 members of 15 Hungarian families with venous thromboembolic diseases and to identify hitherto undescribed genetical defects in the protein C (PROC) gene. The exons of the PROC gene were screened for mutations with the combination of polymerase chain reaction (PCR) and denaturing gradient gel electrophoresis (DGGE), and/or PCR and single-strand conformation polymorphism (SSCP) analysis. The amplified DNA fragments with aberrant migration during DGGE and SSCP were sequenced. Mutations were determined in the PROC gene in all of the examined families: one nonsense mutation, one rare frameshift deletion and nine different missense mutations, of which three were novel (1493 A-->G, 35Asp-->Gly; 6231 G-->A, 173Gly-->Glu; 8476 C-->T, 254Thr-->Ile). The combination of hereditary protein C deficiency with other hereditary thrombophilias was rather common. DGGE and SSCP were proved to be efficient and cost-effective screening methods in the genetic analysis of hereditary protein C deficiency.