Characterization of RHD alleles present in serologically RHD-negative donors determined by a sensitive microplate technique

Autor: Afonso José Pereira Cortez, Flavia Roche Moreira Latini, Janaína Guilhem Muniz, Rosangela de Medeiros Person, Lilian Castilho, Tatiane Aparecida de Paula Vendrame, Carine Prisco Arnoni
Rok vydání: 2019
Předmět:
Zdroj: Vox sanguinisReferences. 114(8)
ISSN: 1423-0410
Popis: Background and objectives Weak D phenotypes with very low antigen densities and DEL phenotype may not be detected in RhD typing routine and could be typed as D-negative, leading to D alloimmunization of D-negative recipients. The present study aimed to investigate the presence of RHD-positive genotypes in blood donors typed as D-negative by an automated system using the solid-phase methodology as a confirmatory test. Methods Two screenings were performed in different selected donor populations. For the first screening, we selected 1403 blood donor samples typed as D-negative regardless of the CE status, and in the second screening, we selected 517 donor samples typed as D-negative C+ and/or E+. RhD typing was performed by microplate in an automated equipment (Neo-Immucor®), and the confirmatory test was performed by solid-phase technique using Capture R® technology. A multiplex PCR specific to RHD and RHDψ was performed in a pool of 6 DNA samples. Sequencing of RHD exons was performed in all RHD-positive samples, and a specific PCR was used to identify the D-CE(4-7)-D hybrid gene. Results and conclusion No weak D type was found in either screening populations. Additionally, 353 (18·4%) D-negative samples presented previously reported non-functional RHD genes, 2 samples had a DEL allele, and 6 samples demonstrated new alleles, including one novel DEL allele. Our study identified six new RHD alleles and showed that the inclusion of a confirmatory test using serological methodology with high sensitivity can reduce the frequency of weak D samples typed as D-negative.
Databáze: OpenAIRE
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