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The routinely used serologic methods are robust in accurately typing standard D− or D+ blood. However, they result in discrepancy in weak or partial D blood, which requires genetic analysis. We have previously used denaturing high‐performance liquid chromatography (DHPLC) to screen the entire RHD‐coding sequence. However, DHPLC is technically challenging, labor‐intensive, and time‐consuming. To overcome these inconveniences, we sought to develop a new two‐step approach.

A total of 430 blood samples with D phenotype ambiguity were recruited for this study. The three most frequent weak D alleles (i.e., weak D, Type 1; weak D, Type 2; and weak D, Type 3), which altogether account for 60% to 90% of the atypical RHD alleles in the Caucasian population, were first identified by Tm‐shift genotyping. The remaining unidentified samples were then …