Abstract: 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.

Key words: RhD, Genotyping, Blood donor