title: Identification of 217 unreported mutations in the F8 gene in a group of 1,410 unselected Italian patients with hemophilia A.
abstract: To provide a National database, 1,410 unrelated hemophilia A (HA) patients were investigated using screening methods denaturing high-performance liquid chromatography (DHPLC), conformational-sensitive gel electrophoresis (CSGE)] and/or direct sequencing. F8 gene mutations were identified in 877 (81%), 146 (82%), and 133 (89%) families with severe, moderate, or mild HA, respectively. Among the 382 different mutations detected, 217 (57%) have not previously been reported in the F8 Haemophilia A Mutation, Structure, Test and Resource Site (HAMSTeRS) database. Mutations leading to a null allele accounted for 82, 15%, and less than 1% of severe, moderate, or mild HA, respectively. A missense mutation was identified in 16%, 68%, and 81% of severe, moderate, or mild HA, respectively. They included 105 missense mutations (48%), 41 small deletions (19%), 25 splice site mutations (12%), 24 nonsense mutations (11%), 18 insertions (8%), three large deletions (1%), and one deletion plus insertion. Unreported mutations were distributed throughout the F8 gene, as they affected all F8 exons but exon 20. We report a wide spectrum of mutations collected in a large National database. The type of mutation was a strong predictor of the clinical phenotype. This database is expected to considerably improve the genetic counseling and medical care of HA families in Italy.
authors.affiliation: Cattedra di Genetica Medica, Dipartimento di Scienze Biomediche, Università degli Studi di Foggia, viale Pinto, Foggia, 71100, Italy.
authors.forename: R
authors.initials: R
authors.lastname: Santacroce
authors.affiliation: Laboratorio di Ematologia ed Emofilia, IV Divisione di Pediatria, Istituto Gaslini, Genova, Italy.
authors.forename: M
authors.initials: M
authors.lastname: Acquila
authors.affiliation: Servizio Trasfusionale, Castelfranco Veneto Hospital (TV), ASL 8 Regione Veneto, Veneto, Italy.
authors.forename: D
authors.initials: D
authors.lastname: Belvini
authors.affiliation: Dipartimento di Biochimica e Biotecnologie Mediche, Università di Napoli "Federico II", CEINGE Biotecnologie Avanzate, Napoli, Italy.
authors.forename: G
authors.initials: G
authors.lastname: Castaldo
authors.affiliation: "Angelo Bianchi Bonomi" Hemophilia and Thrombosis Center, University of Milan and IRCCS Maggiore Hospital, Mangiagalli and regina Elena Foundation, Milan, Italy.
authors.forename: I
authors.initials: I
authors.lastname: Garagiola
authors.affiliation: Divisione di Ematologia, "S. Bortolo" Hospital, Vicenza, Italy.
authors.forename: S H
authors.initials: SH
authors.lastname: Giacomelli
authors.affiliation: Dipartimento di Scienze Mediche e Chirurgiche, Università di Padova, Padova, Italy.
authors.forename: A M
authors.initials: AM
authors.lastname: Lombardi
authors.affiliation: SOD Diagnostica Genetica, Azienda Ospedaliero-Universitaria Careggi Firenze, Firenze, Italy.
authors.forename: B
authors.initials: B
authors.lastname: Minuti
authors.affiliation: UOS di Genetica Molecolare e Citogenetica, Centro di Riferimento Regionale per l'Emofilia e le Malattie Emorragiche Congenite, University Hospital of Parma, Parma, Italy.
authors.forename: F
authors.initials: F
authors.lastname: Riccardi
authors.affiliation: Servizio Trasfusionale, Castelfranco Veneto Hospital (TV), ASL 8 Regione Veneto, Veneto, Italy.
authors.forename: R
authors.initials: R
authors.lastname: Salviato
authors.affiliation: "Angelo Bianchi Bonomi" Hemophilia and Thrombosis Center, University of Milan and IRCCS Maggiore Hospital, Mangiagalli and regina Elena Foundation, Milan, Italy.
authors.forename: L
authors.initials: L
authors.lastname: Tagliabue
authors.affiliation: Unita' di Emostasi e Trombosi, I.R.C.C.S. "Casa Sollievo della Sofferenza", S. Giovanni Rotondo, Italy.
authors.forename: E
authors.initials: E
authors.lastname: Grandone
authors.affiliation: Cattedra di Genetica Medica, Dipartimento di Scienze Biomediche, Università degli Studi di Foggia, viale Pinto, Foggia, 71100, Italy. m.margaglione@unifg.it.|Unita' di Emostasi e Trombosi, I.R.C.C.S. "Casa Sollievo della Sofferenza", S. Giovanni Rotondo, Italy. m.margaglione@unifg.it.
authors.forename: M
authors.initials: M
authors.lastname: Margaglione
mesh_terms: D017398:Alternative Splicing; D002851:Chromatography, High Pressure Liquid; D005091:Exons; D005169:Factor VIII / Q000737:chemistry / Q000235:genetics*; D006467:Hemophilia A / Q000097:blood / Q000235:genetics*; D007558:Italy; D009154:Mutation; D020125:Mutation, Missense; D011487:Protein Conformation; D017384:Sequence Deletion
chemical_list: C078147:F8 protein, human; D005169:Factor VIII