ORIGINAL PAPER FYX real-time polymerase chain reaction with melting curve analysis associated with a complete one-step real-time FY genotyping

• Published in: Vox sanguinis Vol. 92; no. 2; p. 142
• Main Authors: Ansart-Pirenne, H, Martin-Blanc, S, P.-Y. Le Pennec, Rouger, P, J.-P. Cartron, Tournamille, C
• Format: Journal Article
• Language: English
• Published: Amsterdam S. Karger AG 01.02.2007
• Subjects: Black people; Blood transfusions; Genetics; Genotype & phenotype; Medical technology; White people
• ISSN: 0042-9007; 1423-0410
• DOI: 10.1111/j.1423-0410.2006.00872.x

The Duffy (FY) blood group system is controlled by four major alleles: FY*A and FY*B, the Caucasian common alleles, encoding Fya and Fyb antigens; FY*X allele responsible for a poorly expressed Fyb antigen, and FY*Fy a silent predominant allele among Black population. Despite the recent development of a real-time fluorescent polymerase chain reaction (PCR) method for FY genotyping FY*X genotyping has not been described by this method. This study focused on the real-time FY*X genotyping development associated with a complete, one-step real-time FY genotyping, based on fluorescence resonance energy transfer (FRET) technology. Seventy-two blood samples from Fy(a+b-) Caucasian blood donors were studied by real-time PCR only. Forty-seven Caucasian and Black individual blood samples, referred to our laboratory, were studied by PCR-RFLP and real-time PCR. For each individual, the result of the genotype was compared to the known phenotype. The FY*X allele frequency calculated in an Fy(a+b-) Caucasian blood donors population was 0 014. With the Caucasian and Black patient samples we found a complete correlation between PCR-RFLP and the real-time PCR method whatever the alleles combination tested. When the known phenotype was not correlated to FY*X genotype, the presence of the Fyb antigen was always confirmed by adsorption-elution. The real-time technology method is rapid and accurate for FY genotyping. From now, we are able to detect the FY*X allele in all the alleles combinations studied. Regarding its significant frequency, the detection of the FY*X allele is useful for the correct typing of blood donors and recipients considering the therapeutic use of blood units and the preparation of test red blood cells for antibody screening. [PUBLICATION ABSTRACT]