RhD(-)无偿献血者中RhD表型与基因型检测情况分析

Analysis of RhD Phenotype and Genotype in RhD(-)Blood Donors

目的寻找一种有效针对无偿献血者的RhD血型基因定型方法。方法对30例RhD(–)无偿献血者血液样本进行RhD基因定型、弱D检测和Rh CE基因定型。应用荧光定量PCR检测RHD启动子、第4内含子,以及第7、10外显子。结果 30例RhD(–)无偿献血者中,21例未检测到RHD基因,2例为杂交型,其余7例存在RHD启动子、第4内含子和第7、10外显子,其中5例为DEL型,1例为弱D,1例为部分D。30例无偿献血者的Rh CE基因型为,20例C–E–c+e+,6例C+E–c+e–,2例C+E–c+e+,1例C–E+c+e–。结论采用RHCE分型作为Rh(–)定性的筛选检查,能较好地区分D变异体。

Objective To built an effective method for RhD genotyping. Methods RhD phenotyping, weak D testing and RhCE phenotyping were performed in 30 samples from members of the RhD-negative club in Nanjing Red Cross Blood Center. The RhD promoter, intron 4, and exons 7 and 10 were analysed by real-time PCR. Results Of 30 RhD- negative persons, 21 showed complete deletion of the RhD gene, 2 showed the results consistent with RhD-CE-Dhybrid, and 7 showed amplification of RhD promoter, intron 4, and exons 7 and 10. Of the latter group, 5 were in the DEL blood group, 1 were weak D, 1 was partial D. The RhD-negative phenotype samples consisted of 20 C-E-c＋e＋, 6 C＋E-c＋e-, 2 C＋E-c＋e＋ and 1 C-E＋c＋e-. The C-E-c＋e＋ phenotype showed 100% positive predictive value for detecting D-negative cases. Conclusion RhCE phenotyping is efficient to distinguish D-variants in RhD （-） screening test.