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-D hybrid, 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.%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RhCE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RhCE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.
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