title: Rapid identification of female haemophilia A carriers with deletions in the factor VIII gene by quantitative real-time PCR analysis.
abstract: Large deletions of the factorVIII gene account for approximately 5% of severe haemophilia A patients. Although deletions are readily detectable in males, the identification of heterozygosity in possible carriers of these families still constitutes a challenge. In order to identify a deleted allele over the background of the normal allele in these carriers, we developed a rapid real-time quantitative PCR approach by means of LightCycler technology and SYBR green I for monitoring product formation. The method was applied to families with independent deletions (one in exon 14 and the other in exons 23-24) of the Factor VIII gene, thereby allowing a reliable determination of carrier or non-carrier status. The method is extremely versatile and can be adapted to other deletions within the factorVIII gene as well as to other diseases whose molecular pathology consists of deletions or duplications.
authors.affiliation: Genetics, Hospital de Sant Pau Padre Claret 167 08025, Barcelona, Spain. etizzano@hsp.santpau.es
authors.forename: Eduardo F
authors.initials: EF
authors.lastname: Tizzano
authors.forename: María J
authors.initials: MJ
authors.lastname: Barceló
authors.forename: Manel
authors.initials: M
authors.lastname: Baena
authors.forename: Mónica
authors.initials: M
authors.lastname: Cornet
authors.forename: Adoración
authors.initials: A
authors.lastname: Venceslá
authors.forename: José
authors.initials: J
authors.lastname: Mateo
authors.forename: Jordi
authors.initials: J
authors.lastname: Fontcuberta
authors.forename: Montserrat
authors.initials: M
authors.lastname: Baiget
mesh_terms: D001483:Base Sequence; D052160:Benzothiazoles; D003959:Diamines; D005091:Exons; D005169:Factor VIII / Q000235:genetics*; D005260:Female; D006580:Genetic Carrier Screening / Q000379:methods*; D005820:Genetic Testing / Q000379:methods*; D006467:Hemophilia A / Q000175:diagnosis* / Q000235:genetics; D006801:Humans; D008297:Male; D009930:Organic Chemicals; D010375:Pedigree; D016133:Polymerase Chain Reaction; D011804:Quinolines; D015203:Reproducibility of Results; D017384:Sequence Deletion
chemical_list: D052160:Benzothiazoles; D003959:Diamines; D009930:Organic Chemicals; D011804:Quinolines; C098022:SYBR Green I; D005169:Factor VIII