深圳献血人群Duffy血型基因多态性的研究

Polymorphism of Duffy blood group gene in Shenzhen blood donors

目的建立Duffy血型的分子生物学检测方法,研究深圳献血人群Duffy血型基因多态性。方法在2011年8～11月本中心900名无血源关系献血者标本中,用卡式微柱凝胶抗球蛋白方法鉴定Duffy血型表现型,建立Duffy血型的基因分型方法 PCR-SSP法及PCR产物直接测序法,对900例标本DNA进行PCR-SSP法检测,其中50例做Duffy血型基因编码区域序列测定。结果血清学结果为Fy(a+b-)855例,Fy(a+b+)43例,Fy(a-b+)2例,Fy(a-b-)0例。900例Duffy血型PCR-SSP法基因分型结果与血清学表现型完全一致。50例FY基因编码序列部分测序结果与血清学、PCR-SSP法吻合,表现型FY(a+b-)标本测序结果可见FY基因第2外显子第125位核苷酸碱基为纯合子G;表现型FY(a-b+)标本测序结果为第125位核苷酸碱基为纯合子A。结论 Duffy血型PCR-SSP基因分型方法及PCR产物直接测序法可以正确地鉴定Duffy基因型,适用于Duffy血型基因多态性的研究。深圳地区Fya的基因频率为0.973 9,Fyb的基因频率为0.026 1。

Objective To establish molecular method for Duffy blood group genotyping, and to study the polymorphism of Duffy blood group gene in Shenzhen blood donors. Methods A total of 900 blood samples from unrelated blood donors were phenotyped for Fy~ and Fyb by the antiglobulin gel test. PCR-SSP and direct DNA sequencing of the FY gene coding re- gion were established. An of them were detected by PCR-SSP. Fifty DNA samples were studied by direct DNA sequen- cing. Results A total of 855 of them were Fy（ a ＋ b - ） ,43 of them were Fy（ a ＋ b ＋ ）, and 2 of them were Fy（ a - b ＋ ）, respectively. And Fy （ a - b - ） had not been found in our study. The serological results were completely in accordance with FY genotypes by PCR-SSP. The fifty samples sequencing results were consistent with the results via serology trod PCR- SSP. The results of sequencing showed the individuals in Fy（ a ＋ b - ） and Fy（ a - b ＋ ） phenotype differed by a single substitution at nt 125G 〉 A in exon 2 of FY gene. Conclusion DNA-based PCR-SSP and direct DNA sequencing can be correcdy genotyped for FY gene. It will be usefnl in the polymorphism of FY gene research. Fya gene frequency was 0. 973 9, Fyb gene frequency was 0. 026 1 in Shenzhen blood donors.