采用熔解曲线分析法检测红细胞Duffy血型抗原Fy-a/b基因型

Application of Melting Curve to Analyze Genotype of Duffy Blood Group Antigen Fy-a/b

本研究建立用于红细胞Duffy血型抗原Fy-a/b基因分型的熔解曲线分析方法。根据β-actin和Fy-a/bmRNA序列设计并合成引物,建立实时荧光多重PCR-熔解曲线分析法,并用该方法与常规PCR-SSP法同时检测中国成都地区198名献血者Duffy血型抗原Fy-a/b的基因型。结果表明,熔解曲线分析法和常规PCR-SSP法的基因分型结果完全一致。198名中国成都地区献血者中Fya(+)178例(89.9%),Fya(+)Fyb(+)杂合子19例(9.6%),Fyb(+)1例(0.5%),Fya基因频率0.947,Fyb基因频率0.053,符合中国人Duffy血型分布特点。结论:本研究成功建立了红细胞Duffy血型抗原Fy-a/b基因分型的熔解曲线分析方法,该方法适用于大规模Duffy血型基因分型。中国成都地区的献血者Duffy血型基因以Fya为主。

This study was aimed to establish the real-time multiple-PCR with melting curve analysis for Duffy blood group Fy-a/b genotyping. According to the sequence of mRNA coding for fl-actin and Fy-a/b, the primers of fl-actin and Fy-a/b were synthesized. The real-time multiple-PCR with melting curve analysis for Fy-a/b genotyping was established. The Fy-a/b genotyping of 198 blood donors in Chinese Chengdu area has been investigated by melting curve analysis and PCR-SSP. The results showed that the results of Fy-a/b genotype by melting curve analysis were consistent with PCR-SSP. In all of 198 donors in Chinese Chengdu, 178 were Fy^a （ ＋ ） （ 89.9% ）, 19 were Fy^a （ ＋ ） Fy^b （ ＋ ） （9. 6% ）, and 1 was Fyb（ ＋ ） （0.5%）. The gene frequency of Fya was 0.947, while that of Fy^b was 0. 053. It is concluded that the genotyping method of Duffy blood group with melting curve analysis is established, which can be used as a high-throughput screening tool for Duffy blood group genotyping; and the Fya genotype is the major of Duffy blood group of donors in Chinese Chengdu area.