D variants in the population of D-negative blood donors in the north-eastern region of Croatia.
D antigen
D variant
RHD genotyping
partial D
weak D
Journal
Transfusion medicine (Oxford, England)
ISSN: 1365-3148
Titre abrégé: Transfus Med
Pays: England
ID NLM: 9301182
Informations de publication
Date de publication:
Feb 2021
Feb 2021
Historique:
received:
30
04
2020
revised:
16
09
2020
accepted:
22
09
2020
pubmed:
10
10
2020
medline:
3
11
2021
entrez:
9
10
2020
Statut:
ppublish
Résumé
The 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. Altered or reduced D antigen expression can result in D variants, which can be mistyped and can lead to the alloimmunisation of the blood recipient. RHD genotyping can distinguish D variants: weak D, partial D and DEL, thus preventing alloimmunisation. A total of 6523 samples obtained from D-negative Croatian donors were screened for the presence of RHD using 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 the RHD was performed when the genotyping kits failed to assign a D variant. RHD molecular screening revealed 23 (0.35%) RHD-PCR positive samples, all C/E positive, in decreasing frequency: 11 hybrid RHD-CE (2-9) D-CE variants, 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. Data on the significant frequency of D variants among serologically D-negative blood donors in the north-eastern region of Croatia could help in introducing RHD molecular screening of blood donors in a routine workflow.
Sections du résumé
OBJECTIVES
OBJECTIVE
The 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.
BACKGROUND
BACKGROUND
Altered or reduced D antigen expression can result in D variants, which can be mistyped and can lead to the alloimmunisation of the blood recipient. RHD genotyping can distinguish D variants: weak D, partial D and DEL, thus preventing alloimmunisation.
MATERIAL/METHODS
METHODS
A total of 6523 samples obtained from D-negative Croatian donors were screened for the presence of RHD using 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 the RHD was performed when the genotyping kits failed to assign a D variant.
RESULTS
RESULTS
RHD molecular screening revealed 23 (0.35%) RHD-PCR positive samples, all C/E positive, in decreasing frequency: 11 hybrid RHD-CE (2-9) D-CE variants, 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.
CONCLUSION
CONCLUSIONS
Data on the significant frequency of D variants among serologically D-negative blood donors in the north-eastern region of Croatia could help in introducing RHD molecular screening of blood donors in a routine workflow.
Substances chimiques
Rh-Hr Blood-Group System
0
Rho(D) antigen
0
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
43-47Informations de copyright
© 2020 British Blood Transfusion Society.
Références
Daniels G. Variants of RhD - current testing and clinical consequences. Br J Haematol. 2013;161:461-470.
Flegel WA. The genetics of the Rhesus blood group system. Blood Transfus. 2007;5:52-57.
International Society of Blood Transfusion. Table of Blood Group Antigens v.8_180620, 2018. http://www.isbtweb.org/fileadmin/user_upload/Red_Cell_Terminology_and_Immunogenetics/Table_of_blood_group_antigens_within_systems_v8_180620.pdf. Accessed December 20, 2019.
Sandler SG, Flegel WA, Westhoff CM, et al. It´s time to phase in RHD genotyping for patients with a serologic weak D phenotype? College of American Pathologists Transfusion Medicine Resource Committee Work Group. Transfusion. 2015;55:680-689.
Luo X, Keller MA, James I, et al. Strategies to identify candidates for D variant genotyping. Blood Transfus. 2018;16:293-301.
Wagner FF. Getting comfortable with RH blood group system terminologies and database. Vox Sang. 2018;113(suppl. 1):11-12.
Kim KH, Kim KE, Woo KS, Han JY, Kim JM, Park KU. Primary anti-D immunization by DEL red blood cells. Korean J Lab Med. 2009;29:361-365.
Wagner FF, Frohmajer A, Flegel WA. RHD positive haplotypes in D negative Europeans. BMC Genet. 2001;2:10.
Yasuda H, Ohto H, Sakuma S, Ishikawa Y. Secondary anti-D immunization by Del red blood cells. Transfusion. 2005;45:1581-1584.
Christiansen M, Sorensen BS, Grunnet N. RHD positive among C/E+ and D-blood donors in Denmark. Transfusion. 2010;50:1460-1464.
Crottet SL, Henny C, Meyer S, et al. Implementation of a mandatory donor RHD screening in Switzerland. Transfus Apher Sci. 2014;50:169-174.
Flegel WA, von Zabern I, Wagner FE. Six years' experience performing RHD genotyping to confirm D- red blood cell units in Germany for preventing anti D-immunizations. Transfusion. 2009;49:465-471.
Wagner FF. RHD PCR of D-negative blood donors. Transfus Med Hemother. 2013;40:172-181.
Hromadnikova I, Vechetova L, Vesela K, et al. Non-invasive fetal RHD exon 7 and exon 10 genotyping using real-time PCR testing of fetal DNA in maternal plasma. Fetal Diagn Ther. 2005;20:275-280.
Legler TJ, Maas JH, Köhler M, et al. RHD sequencing, a new tool for decision making on transfusion therapy and provision of Rh prophylaxis. Transfus Med. 2001;11:383-388.
Polin H, Danzer M, Gasznern W, et al. Identification of RHD alleles with the potential of anti-D immunization among seemingly D-blood donors in Upper Austria. Transfusion. 2009;49:676-681.
Wagner T, Körmöczi GF, Buchta C, et al. Anti-D immunization by DEL red blood cells. Transfusion. 2005;45:520-526.
Dogic V, Bingulac-Popovic J, Babic I, et al. Distribution of weak D types in the Croatian population. Transfus Med. 2011;21:278-279.
Gassner C, Doescher A, Drnovsek TD, et al. Presence of RHD in serologically D-, C/E+ individuals, a European multicenter study. Transfusion. 2005;45:527-538.
Hussein E, Teruya J. Serologic findings of RhD alleles in Egyptians and their clinical implications. Transfus Apher Sci. 2014;51:184-187.
Moussa H, Tsochandaridis M, Chakroun T, et al. Molecular background of D-negative phenotype in the Tunisian population. Transfus Med. 2012;22:192-198.
Perez-Alvarez I, Hayes C, Hailemariam T, Shin E, Hutchinson T, Klappern E. RHD genotyping of serologic RhD-negative blood donors in a hospital-based blood donor center. Transfusion. 2019;59:2422-2428.
Gowland P, Gassner C, Hustinx H, et al. Molecular RHD screening of RhD negative donors can replace standard serological testing for RhD negative donors. Transfus Apher Sci. 2014;50:163-168.
Seo MH, Won EJ, Hong YJ, et al. An effective diagnostic strategy for accurate detection of RhD variants including Asian DEL type in apparently RhD-negative blood donors in Korea. Vox Sang. 2016;111:425-430.
Orzinska A, Guz K, Polin H, et al. RHD variants in Polish blood donors routinely typed as D−. Transfusion. 2013;2013(53):2945-2953.
Körmöczi GF, Gassner C, Shao CP, Uchikawa M, Legler TJ. A comprehensive analysis of DEL types, partial DEL individuals are prone to anti-D alloimmunization. Transfusion. 2005;45(10):1561-1567.
Kwon DH, Sandler SG, Flegel WA. DEL phenotype. Immunohematology. 2017;33:125-132.
Scott SA, Nagl L, Tilley L, et al. The RHD(1227G>A) DEL-associated allele is the most prevalent DEL allele in Australian D- blood donors with C+ and/or E+ phenotypes. Transfusion. 2014;54:2931-2940.