Quantitative Real-Time PCR Assay for Rapid Identification of Deletion Carriers in Hemophilia

Gene deletions are common events in various hereditary diseases, but conventional PCR often fails to detect such defects in heterozygous patients (1). This highlights the need for a suitable method, easily applicable in a diagnostic laboratory (2). Large deletions account for ∼5% of cases of hemophilia, inherited as a recessive X-linked disease, making determination of carrier status difficult. It is of particular importance when a pregnant woman is carrying a male fetus. Prenatal testing that uses invasive methods of sample collection carries a risk of miscarriage or fetal injury, and is unnecessary if the woman is not a carrier. We designed a real-time quantitative PCR assay to establish heterozygous status for deletion carriers and to avoid such risky situations in a family in which a large deletion of the F9 gene was responsible for a severe …