title: Conformation sensitive gel electrophoresis for detection of factor X gene mutations.
abstract: Fifteen patients from five families with laboratory data suggesting factor X (FX) deficiency were screened for causative mutations by conformation sensitive gel electrophoresis (CSGE) followed by sequencing. All exonic and flanking intronic regions of factor X gene were amplified using PCR. After heteroduplex formation, samples were analyzed onto a polyacrylamide gel for possible mismatch. An abnormal CSGE profile indicating an heteroduplex was identified in 10/15 cases. All the 10 patients with a patter of migration suggesting a mismatch had a laboratoristic pattern of FX deficiency whereas the five cases with a normal CSGE aspect referred to the normal components of the families who did not carry any FX defect. Sequencing demonstrated that the 10 exons, which showed a suspect CSGE pattern, harbored a mutation responsible for the factor X defect. Of the five mutation identified, two were recognized to be novel mutations (a 871C>T substitution and a 1169G>T transversion in exon 8), both located in the catalytic portion of FX. CSGE may be an effective and simple procedure for screening factor X gene mutations.
authors.affiliation: Department of Medical and Surgical Sciences, Second Chair of Medicine, University of Padua Medical School, via Ospedale, 105, 35100, Padua, Italy. fabvian@unipd.it
authors.forename: Fabrizio
authors.initials: F
authors.lastname: Vianello
authors.forename: Anna Maria
authors.initials: AM
authors.lastname: Lombardi
authors.forename: Federico
authors.initials: F
authors.lastname: Dal Bello
authors.forename: Ezio
authors.initials: E
authors.lastname: Zanon
authors.forename: Laura
authors.initials: L
authors.lastname: Cabrio
authors.forename: Antonio
authors.initials: A
authors.lastname: Girolami
mesh_terms: D000293:Adolescent; D000328:Adult; D000368:Aged; D001780:Blood Coagulation Tests; D020134:Catalytic Domain / Q000235:genetics; D002648:Child; D002675:Child, Preschool; D004252:DNA Mutational Analysis / Q000379:methods; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D005170:Factor X / Q000737:chemistry / Q000235:genetics*; D005171:Factor X Deficiency / Q000175:diagnosis* / Q000235:genetics; D005192:Family Health; D005820:Genetic Testing / Q000379:methods; D020180:Heteroduplex Analysis; D006801:Humans; D008875:Middle Aged; D009154:Mutation; D010641:Phenotype
chemical_list: D005170:Factor X