BACKGROUND: Antigens in the human red cell Miltenberger series are glycophorin variants of the MN (MNS) blood group system that are due to the rearrangement of glycophorin A (GPA) and glycophorin B (GPB) genes. STUDY DESIGN AND METHODS: Taking advantage of the differences between the GPA and GPB genes, a polymerase chain reaction-based method was developed to detect all the Miltenberger glycophorin variants and St(a) subtype. GPA- and GPB-specific primers were used to amplify the GPA or GPB gene, and the amplified products were used to recognize the different hybrid genes after restriction enzyme digestions. RESULTS: Among 264 Taiwanese subjects studied, Mi.III and St(a) are the most common types of Miltenberger variants found. Mi.III was present in 13 (4.92%) of 264, and St(a) was found in 8 (3.03%) of 264; 1 case (0.4%) of Mi.V was also identified from the study group. CONCLUSION: This is the first polymerase chain reaction-based method of detecting most of the Miltenberger variants and St(a). The genomic typing results were confirmed by control DNA of identified Miltenberger phenotypes. The prevalence rates of Mi.III and St(a) in this study were also consistent with other previous reports using different methods.