Tizzano Eduardo F, Barceló María J, Baena Manel, Cornet Mónica, Venceslá Adoración, Mateo José, Fontcuberta Jordi, Baiget Montserrat
Genetics, Hospital de Sant Pau Padre Claret 167 08025, Barcelona, Spain.
Thromb Haemost. 2005 Sep;94(3):661-4. doi: 10.1160/TH05-03-0144.
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.
凝血因子VIII基因的大片段缺失约占重症A型血友病患者的5%。虽然在男性中很容易检测到缺失，但在这些家族的可能携带者中鉴定杂合性仍然是一项挑战。为了在这些携带者的正常等位基因背景下鉴定缺失的等位基因，我们利用LightCycler技术和SYBR Green I开发了一种快速实时定量PCR方法，用于监测产物形成。该方法应用于凝血因子VIII基因存在独立缺失（一个在外显子14，另一个在外显子23-24）的家族，从而能够可靠地确定携带者或非携带者状态。该方法具有极高的通用性，可适用于凝血因子VIII基因内的其他缺失，以及分子病理学由缺失或重复组成的其他疾病。
