D variants in the population of D‐negative blood donors in the north‐eastern region of Croatia

• DOI: 10.1111/tme.12726
• Publication Date: 2020-10-09T09:34:13Z
• Authors: Hana Safic Stanic; Vesna Dogic; Ivona Herceg; Sandra Jagnjic; Jasna Bingulac‐Po...; Ivana Babic; Ante Corusic; Irena Jukic
• Keywords: Adult; Male; Rh-Hr Blood-Group System; Adolescent; Genotype; Genotyping Techniques; Croatia; Blood Donors; Exons; Middle Aged; Real-Time Polymerase Chain Reaction; 3. Good health; 03 medical and health sciences; 0302 clinical medicine; D antigen ; D variant ; RHD genotyping ; partial D ; weak D; Humans; Female; Polymorphism, Single-Stranded Conformational; Aged

AbstractObjectivesThe aim of this study was to determine RHESUS D GENE (RHD)allelic variants among Croatian D‐negative blood donors and compare our results with respective data from other European countries.BackgroundAltered or reduced D antigen expression can result in D variants, which can be mistyped and can lead to the alloimmunisation of the blood recipient.RHDgenotyping can distinguish D variants: weak D, partial D and DEL, thus preventing alloimmunisation.Material/methodsA total of 6523 samples obtained from D‐negative Croatian donors were screened for the presence ofRHDusing the real‐time polymerase chain reaction (PCR) method. PCR‐SSP was performed for D variant genotyping by using commercial genotyping kits (Inno‐Train, Kronberg, Germany). Genomic DNA sequencing for all 10 exons of theRHDwas performed when the genotyping kits failed to assign a D variant.ResultsRHDmolecular screening revealed 23 (0.35%)RHD‐PCR positive samples, all C/E positive, in decreasing frequency: 11 hybridRHD‐CE (2‐9) D‐CEvariants, 4 weak partial D type 11 and 2 weak D type 2. Six samples remained unresolved and were sequenced. For 12 of 23 samples (excluding large hybrids), an adsorption/elution of anti‐D serum was performed, confirming that all 12 were RhD+. The calculated frequency of clinically significant D alleles in RhD‐negative blood donors was 1:543 (0.18%) or 1:53 (1.89%) in C/E blood donors.ConclusionData on the significant frequency of D variants among serologically D‐negative blood donors in the north‐eastern region of Croatia could help in introducingRHDmolecular screening of blood donors in a routine workflow.