Molecular basis of weak D expression in the Indian population and report of a novel, predominant variant RHD allele.

BACKGROUND: The Rh blood group system is the most polymorphic system and is implicated in hemolytic transfusion reaction and hemolytic disease of the fetus and newborn. Molecular genetics of the RH genes have been extensively studied in Caucasians, Africans, and East Asians and the variant alleles giving rise to weak and partial D phenotypes have been reported. However, limited genetic studies have been carried out in the large Indian population, even though the variability of Rh expression has been documented. STUDY DESIGN AND METHODS: In this study we sought to characterize the molecular bases of weak D expression in Indians. RHD gene in samples presenting with a weak D phenotype by serologic analyses (n = 223) was genotyped by conventional molecular approaches. RESULTS: In addition to referenced and novel single-nucleotide variations, a novel approximately 12-kb duplication event, including Exon 3, was identified predominantly in variant D samples (130/223, 58.3%) and characterized at the nucleotide sequence level. Functional analyses suggested that this genetic variation quantitatively affects the expression of the normal transcript and then subsequently the expression of the normal RhD protein. CONCLUSION: We describe a major novel, variant RHD allele in Indians that can be easily identified routinely by implementing a simple genotyping assay. Although we may consider this variation as a weak partial D variant, further studies and observations are needed to confirm the same. These findings may contribute to improve significantly Rh blood group diagnostics in more than one billion Indians.

Screening for DEL phenotype in RhD negative Indians.

BACKGROUND: DEL phenotype represents a very weak form of D variant detected only by adsorption and elution technique. DEL phenotype individuals mistyped as RhD-negative can lead to alloimmunization after transfusion or pregnancy. Molecular techniques have now been used to identify DEL variants. They are commonly encountered in the East Asian population with RHD(K409K) being the most frequent allele. RHD(M295I) is the most common DEL allele in Caucasians. As there is a paucity of data on DEL phenotype in the Indian population, the study aims to screen RhD negative individuals for two most common DEL mutations. MATERIAL AND METHODS: EDTA blood was collected from 900 RhD negative individuals. Serological analysis included testing for the five major Rh antigens- C, c, D, E, and e by tube technique. Samples showing negative reaction for the presence of D antigen by Indirect Antiglobulin test were further tested for DEL phenotype by adsorption and elution technique. Molecular analysis involved DNA extraction and testing by PCR-SSP for RHD(K409K) and RHD(M295I) DEL alleles. RESULTS: Rh phenotyping showed 153 Rh negative individuals with r'r, ten with r''r and 737 with rr phenotype. All the samples tested negative for RhD antigen by adsorption and elution method. The two common DEL mutations RHD(K409K) and RHD(M295I) were also not detected in the study population. CONCLUSION: The study population showed the absence of the two common DEL alleles, concluding the variant to be rare. A comprehensive study with a larger sample size to look for other DEL mutations should be performed.