Thromb Haemost 2005; 94(03): 661-664
DOI: 10.1160/TH05-03-0144
New Technologies and Diagnostic Tools
Schattauer GmbH

Rapid identification of female haemophilia A carriers with deletions in the factor VIII gene by quantitative Real-Time PCR analysis

Eduardo F. Tizzano
,
María J. Barceló
,
Manel Baena
,
Mónica Cornet
,
Adoración Venceslá
,
José Mateo
,
Jordi Fontcuberta
,
Montserrat Baiget

Financial support: This work was supported by Fundació Catalana d′Hemofilia.
Further Information

Publication History

Received: 01 March 2005

Accepted after major revision: 12 May 2005

Publication Date:
07 December 2017 (online)

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Summary

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 factor VIII gene as well as to other diseases whose molecular pathology consists of deletions or duplications.