Original Article

The RHD Genotyping of Rh-Negative and Weak D Phenotype among Blood Donors in Southeast Iran

RHD genotyping

Abstract

Background: The D antigen is a subset of Rh blood group antigens involved in the hemolytic disease of the newborn [HDFN] and hemolytic transfusion reaction [HTR]. The hybrid Rhesus box that was created after RH gene deletion, was known as a mechanism of the Rh-negative phenotype. Hybrid marker identification is used to confirm the deletion of the RHD gene and to determine zygosity. This study aims to detect this marker in Rh-negative and weak D phenotype blood donors of the southeast of Iran.

Materials and Methods: The molecular analysis of the hybrid Rhesus box was performed on the 200 Rh-negative blood donors in Sistan and Baluchestan province, southeast Iran. The presence of alleles responsible for the D variants was assessed by DNA sequencing in 26 weak D phenotype donors.

Results: Of the 200 Rh-negative blood samples, 198 samples were homozygous (99%), and two samples were heterozygous (1%). Heterozygous samples had RHD*01N.73 allele and the RHD*01N.18 allele. Of the 26 samples with weak D phenotype, 16 partial DLO (61%), 4 partial DBT1 (15.3%), 2 partial DV type 2 (7.7%), 1 weak D type 1, 1 weak D type 4.2.3, 1weak D type 105 and 1 RHD (S103P) (4%) were determined.

Conclusion: Since RHD gene deletion is the main mechanism of the Rh-negativity in Sistan and Baluchestan provinces, a hybrid Rhesus box marker can be used in resolving RhD typing discrepancies by RHD genotyping methods.

1. Levine P, Stetson RE. An unusual case of intra-group agglutination. JAMA. 1984;251(10):1316-7.
2. Agre P, Cartron JP. Molecular biology of the Rh antigens. Blood. 1991 1;78(3):551-63.
3. Avent ND, Reid ME. The Rh blood group system: a review. Blood. 2000;95(2):375-87.
4. Cotorruelo C, Biondi C, Borras SG, et al. Early detection of RhD status in pregnancies at risk of hemolytic disease of the newborn. Clin Exp Med. 2002; 2(2):77-81.
5. Sassi A, Ouchari M, Houissa B, et al. RHD genotyping and its implication in transfusion practice. Transfus Apher Sci. 2014; 51(3):59-63.
6. Sandler SG, Chen LN, Flegel WA. Serological weak D phenotypes: a review and guidance for interpreting the RhD blood type using the RHD genotype. Br J Haematol. 2017;179(1):10-19.
7. Wagner FF, Flegel WA. RHD gene deletion occurred in the Rhesus box. Blood. 2000;95(12):3662-8.
8. Flegel WA. Molecular genetics and clinical applications for RH. Transfu Apher Sci. 2011;44(1):81-91.
9. Flegel WA. Molecular genetics of RH and its clinical application. Transfus Clin Biol. 2006;13(1-2):4-12.
10. Flegel WA, Von Zabern I, Wagner FF. Six years' experience performing RHD genotyping to confirm D− red blood cell units in Germany for preventing anti‐D immunizations. Transfusion. 2009;49(3):465-71.
11. Dezan MR, Oliveira VB, Gomes ÇN et al. High frequency of variant RHD genotypes among donors and patients of mixed origin with serologic Weak‐D phenotype. J Clin Lab Anal. 2018;32(9): e22596.
12. Daniels G. Variants of RhD–current testing and clinical consequences. Br J Haematol. 2013;161(4):461-70.
13. Garratty G. Do we need to be more concerned about weak D antigens?. Transfusion. 2005; 45(10):1547-51.
14. Kulkarni S, Parchure DS, Gopalkrishnan V, et al. Screening for DEL phenotype in RhD negative Indians. J Clin Lab Anal. 2018;32(3): e22288.
15. Perco P, Shao CP, Mayr WR, et al. Testing for the D zygosity with three different methods revealed altered Rhesus boxes and a new weak D type. Transfusion. 2003;43(3):335-9.
16. Ahmadi MH, Hantuoshzadeh S, Okhovat MA, et al. Fetal RHD Genotyping from Circulating Cell-Free Fetal DNA in Plasma of Rh Negative Pregnant Women in Iran. Indian J Hematol Blood Transfus. 2016;32(4):447-453.
17. Doescher A, Flegel WA, Petershofen EK, et al. Weak D type 1.1 exemplifies another complexity in weak D genotyping. Transfusion. 2005;45(10):1568-73.
18. Khosroshahi BN, Oodi A, Namjou S, et al. RHD Genotyping by Molecular Analysis of Hybrid Rhesus box in RhD-Negative Blood Donors from Iran. Indian J Hematol Blood Transfus. 2019;35(1):119-24.
19. Li Q, Hou L, Guo ZH, et al. Molecular basis of the RHD gene in blood donors with DEL phenotypes in Shanghai. Vox San. 2009;97(2):139-46.
20. Johnsen JM. Using red blood cell genomics in transfusion medicine. Hematology Am Soc Hematol Educ Program . 2015;2015:168-76.
21.Taeo M, Jeon S, Yu H, et al. Case of D-Variant from a Frameshift Mutation RHD 711delC. Korean J Blood Transfus. 2019;30(2):168-73.
22. Haer‐Wigman L, Veldhuisen B, Jonkers R, et al. RHD and RHCE variant and zygosity genotyping via multiplex ligation–dependent probe amplification. Transfusion. 2013;53(7):1559-74.
23. Sandler SG, Flegel WA, Westhoff CM, et al. It's time to phase in RHD genotyping for patients with a serologic weak D phenotype. Transfusion. 2015;55(3):680-9.
24. Van Sandt VS, Gassner C, Emonds MP, et al. RHD variants in F landers, B elgium. Transfusion. 2015; 55(6pt2):1411-7.
25. Laget L, Izard C, Durieux-Roussel E, et al. Relevance and costs of RHD genotyping in women with a weak D phenotype. Transfus Clin Biol. 2019;26(1):27-31.
26. Ansart‐Pirenne H, Asso‐Bonnet M, Pennec PYL, et al. RhD variants in Caucasians: consequences for checking clinically relevant alleles. Transfusion. 2004;44(9):1282-6.
27. Lejon Crottet S, Haer‐Wigman L, Gowland P, et al. Serologic and molecular investigations of DAR 1 (weak D Type 4.2), DAR 1.2, DAR 1.3, DAR 2 (DARE), and DARA. Transfusion. 2013;53(11pt2):3000-8.
28. Wallace M, Lomas‐Francis C, Beckers E, et al. DBT: a partial D phenotype associated with the low‐incidence antigen Rh32. Transfus Med. 1997;7(3):233-8.
29. Arnoni CP, Latini FRM, Muniz JG, et al. How do we identify RHD variants using a practical molecular approach? Transfusion. 2014;54(4):962-9.
30. Hussein E, Teruya J. Serologic findings of RhD alleles in Egyptians and their clinical implications. Transfus Apher Sci. 2014;51(2):184-7.
31. Kulkarni S, Colah R, Gorakshakar A, et al. Frequency of partial D in Western India. Transfus Med. 2008;18(2):91-6.
32. Ogasawara K, Sasaki K, Isa K, et al. Weak D alleles in Japanese: a c. 960G> A silent mutation in exon 7 of the RHD gene that affects D expression. Vox Sang. 2016;110(2):179-84.
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IssueVol 15, No 4 (2021) QRcode
SectionOriginal Article(s)
DOI https://doi.org/10.18502/ijhoscr.v15i4.7476
Keywords
Hybrid Rhesus box; D variant;RHD gene deletion; RhD-negative phenotype; Weak D

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How to Cite
1.
Sadeghi-Bojd Y, Amirizadeh N, Oodi A. The RHD Genotyping of Rh-Negative and Weak D Phenotype among Blood Donors in Southeast Iran. Int J Hematol Oncol Stem Cell Res. 2021;15(4):213-220.