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<p>Limited data are available on the molecular basis of D antigen negativity in eastern India. Thus, we decided to explore the genetic landscape of D–, C/E+ phenotypes among blood donors by using standard <i>RHD</i> and <i>RHCE</i> genotyping approaches, which could provide valuable insight to understand the genetic basis of D antigen negativity in this population. In this single-center observational study, we tested whole blood donors over four years in Kolkata. The RhD type, weak D status, and CcEe phenotypes were determined using the standard serological method. An adsorption-elution test was performed to check for the Del phenotype. We conducted <i>RHD</i> genotyping using PCR-SSP by targeting exons 1, 5, and 10, followed by next-generation amplicon sequencing when necessary. A total of 56,445 whole blood donors were tested. Among them, 2,421 were confirmed as D– (4.29%). We found 194 D-, C/E+ samples. Complete deletion of the <i>RHD</i> gene was observed in 178 cases (91.8%), whereas 16 (8.2%) carried an <i>RHD</i> allele, indicating the presence of an <i>RHD-CE-D</i> hybrid box. Further testing revealed 14 donors were carrying the <i>RHD-CE(2–9)-D</i> allele <i>(RHD*01&#xa0;N.03)</i>, while the <i>RHD-CE(2–7)-D</i> allele (<i>RHD*01N.05</i>) was identified in two donors. Overall, <i>RHD-CE-D</i> hybrid alleles are not highly prevalent in D–, C/E+ blood donors in Eastern India. Western recommendations of <i>RHD</i> genotyping for all D– donors should be interpreted cautiously in the Indian population. The <i>RHD</i> genotyping strategy effectively predicts <i>RHD-CE-D</i> hybrid alleles, particularly when there is an absence of PCR product for <i>RHD</i> exon 5.</p>

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The Genetic Landscape of RhD-Negative Blood Donors with C and/or E-Positive Phenotypes in Eastern India

  • 期刊论文

  • 发表日期：

    2026年4月27日

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  • Najla Haneefa Basheela,
  • Ayesha Sinha,
  • Durba Biswas,
  • Debapriya Basu,
  • Suvro Sankha Datta

摘要

Limited data are available on the molecular basis of D antigen negativity in eastern India. Thus, we decided to explore the genetic landscape of D–, C/E+ phenotypes among blood donors by using standard RHD and RHCE genotyping approaches, which could provide valuable insight to understand the genetic basis of D antigen negativity in this population. In this single-center observational study, we tested whole blood donors over four years in Kolkata. The RhD type, weak D status, and CcEe phenotypes were determined using the standard serological method. An adsorption-elution test was performed to check for the Del phenotype. We conducted RHD genotyping using PCR-SSP by targeting exons 1, 5, and 10, followed by next-generation amplicon sequencing when necessary. A total of 56,445 whole blood donors were tested. Among them, 2,421 were confirmed as D– (4.29%). We found 194 D-, C/E+ samples. Complete deletion of the RHD gene was observed in 178 cases (91.8%), whereas 16 (8.2%) carried an RHD allele, indicating the presence of an RHD-CE-D hybrid box. Further testing revealed 14 donors were carrying the RHD-CE(2–9)-D allele (RHD*01 N.03), while the RHD-CE(2–7)-D allele (RHD*01N.05) was identified in two donors. Overall, RHD-CE-D hybrid alleles are not highly prevalent in D–, C/E+ blood donors in Eastern India. Western recommendations of RHD genotyping for all D– donors should be interpreted cautiously in the Indian population. The RHD genotyping strategy effectively predicts RHD-CE-D hybrid alleles, particularly when there is an absence of PCR product for RHD exon 5.