Transitioning a multiethnic donor pool from serologic D-negative to molecularly RHD-negative at a hospital-based blood donor service

Abstract Background Some individuals carry a very low expression of the D-antigen, called a Del phenotype. Red cell units from such blood donors with DEL alleles are RhD protein-positive, despite being routinely labelled D-negative. Molecular typing offers a more sensitive method to identify Del ind...

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Main Authors: Willy A. Flegel, Kshitij Srivastava, Lorraine G. Caruccio, Pirmin Schmid, David A. Stiles, Marina U. Bueno, Nadine R. Dowling, Traci D. Paige, Sita Shrestha
Format: Article
Language:English
Published: BMC 2025-06-01
Series:Journal of Translational Medicine
Online Access:https://doi.org/10.1186/s12967-025-06716-8
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author Willy A. Flegel
Kshitij Srivastava
Lorraine G. Caruccio
Pirmin Schmid
David A. Stiles
Marina U. Bueno
Nadine R. Dowling
Traci D. Paige
Sita Shrestha
author_facet Willy A. Flegel
Kshitij Srivastava
Lorraine G. Caruccio
Pirmin Schmid
David A. Stiles
Marina U. Bueno
Nadine R. Dowling
Traci D. Paige
Sita Shrestha
author_sort Willy A. Flegel
collection DOAJ
description Abstract Background Some individuals carry a very low expression of the D-antigen, called a Del phenotype. Red cell units from such blood donors with DEL alleles are RhD protein-positive, despite being routinely labelled D-negative. Molecular typing offers a more sensitive method to identify Del individuals by detecting the presence of the RHD gene. Pools of 20 or more donor samples are routinely screened for the RHD gene in some, mostly European, donor populations. Methods A modular real-time PCR assay targeting RHD intron 4, exon 5, and exon 7 was developed for individual testing. We screened for the RHD gene among all blood donors who typed D-negative in routine serology. Results Over 15 years, 2254 D-negative donors were individually tested for the RHD gene. With a sensitivity of detecting 5 RHD positive gDNA copies per reaction, 42 donors tested positive (1.9%). Among them, 34 carried the common RHDΨ allele (80.9%), while 7 harbored 5 known RHD alleles, and 1 a novel RHD deletion. We inadvertently detected 2 other donors with DVI, establishing a population frequency of 1 in 731 for the U.S. Conclusions A modular approach for RHD screening is suitable for blood donors when sample pooling is not feasible among multiethnic donor populations. We transitioned donors since 2009 from serologic D-negative to molecularly RHD-negative status at the NIH Clinical Center. Molecular RHD screening of serologic D-negative donors is an effective way to identify individuals harboring DEL alleles that can cause alloimmunization in transfusion recipients.
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spelling doaj-art-dd2d89874aba4d1097bf7059b310d7fc2025-08-20T03:22:54ZengBMCJournal of Translational Medicine1479-58762025-06-0123111110.1186/s12967-025-06716-8Transitioning a multiethnic donor pool from serologic D-negative to molecularly RHD-negative at a hospital-based blood donor serviceWilly A. Flegel0Kshitij Srivastava1Lorraine G. Caruccio2Pirmin Schmid3David A. Stiles4Marina U. Bueno5Nadine R. Dowling6Traci D. Paige7Sita Shrestha8Laboratory Services Section, Department of Transfusion Medicine, NIH Clinical Center, National Institutes of HealthLaboratory Services Section, Department of Transfusion Medicine, NIH Clinical Center, National Institutes of HealthLaboratory Services Section, Department of Transfusion Medicine, NIH Clinical Center, National Institutes of HealthLaboratory Services Section, Department of Transfusion Medicine, NIH Clinical Center, National Institutes of HealthLaboratory Services Section, Department of Transfusion Medicine, NIH Clinical Center, National Institutes of HealthLaboratory Services Section, Department of Transfusion Medicine, NIH Clinical Center, National Institutes of HealthLaboratory Services Section, Department of Transfusion Medicine, NIH Clinical Center, National Institutes of HealthLaboratory Services Section, Department of Transfusion Medicine, NIH Clinical Center, National Institutes of HealthLaboratory Services Section, Department of Transfusion Medicine, NIH Clinical Center, National Institutes of HealthAbstract Background Some individuals carry a very low expression of the D-antigen, called a Del phenotype. Red cell units from such blood donors with DEL alleles are RhD protein-positive, despite being routinely labelled D-negative. Molecular typing offers a more sensitive method to identify Del individuals by detecting the presence of the RHD gene. Pools of 20 or more donor samples are routinely screened for the RHD gene in some, mostly European, donor populations. Methods A modular real-time PCR assay targeting RHD intron 4, exon 5, and exon 7 was developed for individual testing. We screened for the RHD gene among all blood donors who typed D-negative in routine serology. Results Over 15 years, 2254 D-negative donors were individually tested for the RHD gene. With a sensitivity of detecting 5 RHD positive gDNA copies per reaction, 42 donors tested positive (1.9%). Among them, 34 carried the common RHDΨ allele (80.9%), while 7 harbored 5 known RHD alleles, and 1 a novel RHD deletion. We inadvertently detected 2 other donors with DVI, establishing a population frequency of 1 in 731 for the U.S. Conclusions A modular approach for RHD screening is suitable for blood donors when sample pooling is not feasible among multiethnic donor populations. We transitioned donors since 2009 from serologic D-negative to molecularly RHD-negative status at the NIH Clinical Center. Molecular RHD screening of serologic D-negative donors is an effective way to identify individuals harboring DEL alleles that can cause alloimmunization in transfusion recipients.https://doi.org/10.1186/s12967-025-06716-8
spellingShingle Willy A. Flegel
Kshitij Srivastava
Lorraine G. Caruccio
Pirmin Schmid
David A. Stiles
Marina U. Bueno
Nadine R. Dowling
Traci D. Paige
Sita Shrestha
Transitioning a multiethnic donor pool from serologic D-negative to molecularly RHD-negative at a hospital-based blood donor service
Journal of Translational Medicine
title Transitioning a multiethnic donor pool from serologic D-negative to molecularly RHD-negative at a hospital-based blood donor service
title_full Transitioning a multiethnic donor pool from serologic D-negative to molecularly RHD-negative at a hospital-based blood donor service
title_fullStr Transitioning a multiethnic donor pool from serologic D-negative to molecularly RHD-negative at a hospital-based blood donor service
title_full_unstemmed Transitioning a multiethnic donor pool from serologic D-negative to molecularly RHD-negative at a hospital-based blood donor service
title_short Transitioning a multiethnic donor pool from serologic D-negative to molecularly RHD-negative at a hospital-based blood donor service
title_sort transitioning a multiethnic donor pool from serologic d negative to molecularly rhd negative at a hospital based blood donor service
url https://doi.org/10.1186/s12967-025-06716-8
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