Ansart-Pirenne H, Martin-Blanc S, Le Pennec P-Y, Rouger P, Cartron J-P, Tournamille C
Institut National de la Transfusion Sanguine, Centre National de Référence pour les Groupes Sanguins, 20 rue Bouvier, 75522 Paris cedex 11, France.
Vox Sang. 2007 Feb;92(2):142-7. doi: 10.1111/j.1423-0410.2006.00872.x.
The Duffy (FY) blood group system is controlled by four major alleles: FYA and FYB, the Caucasian common alleles, encoding Fy(a) and Fy(b) antigens; FYX allele responsible for a poorly expressed Fy(b) antigen, and FYFy a silent predominant allele among Black population. Despite the recent development of a real-time fluorescent polymerase chain reaction (PCR) method for FY genotyping FYX genotyping has not been described by this method. This study focused on the real-time FYX 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 FYX 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 FYX genotype, the presence of the Fy(b) 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 FYX allele in all the alleles combinations studied. Regarding its significant frequency, the detection of the FYX 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.
达菲（FY）血型系统由四个主要等位基因控制：FYA和FYB，这是白种人中常见的等位基因，分别编码Fy(a)和Fy(b)抗原；FYX等位基因导致Fy(b)抗原表达较弱，而FYFy是黑人人群中占主导的沉默等位基因。尽管最近开发了一种用于FY基因分型的实时荧光聚合酶链反应（PCR）方法，但该方法尚未用于FYX基因分型。本研究聚焦于基于荧光共振能量转移（FRET）技术，开发与完整的一步式实时FY基因分型相关的实时FYX基因分型。
仅通过实时PCR研究了72份来自Fy(a+b-)白种人献血者的血样。对送至我们实验室的47份白种人和黑人个体血样进行了PCR-RFLP和实时PCR研究。对于每个个体，将基因型结果与已知表型进行比较。
在Fy(a+b-)白种人献血者群体中计算出的FYX等位基因频率为0.014。对于白种人和黑人患者样本，无论测试的等位基因组合如何，我们发现PCR-RFLP与实时PCR方法之间完全相关。当已知表型与FYX基因型不相关时，通过吸附洗脱总能确认Fy(b)抗原的存在。
实时技术方法用于FY基因分型快速且准确。从现在起，我们能够在所研究的所有等位基因组合中检测到FYX等位基因。鉴于其显著的频率，考虑到血液单位的治疗用途以及用于抗体筛查的测试红细胞的制备，检测FYX等位基因对于正确分型献血者和受血者很有用。
