Identification of the RHD novel allele c. 801+2T>G and study of its effect on RhD phenotype in vitro
Journal Title: Chinese Journal of Blood Transfusion - Year 2024, Vol 37, Issue 12
Abstract
[Objective] To further identify the RhD phenotype and RHD genotype in the individual who have RhD negative phenotype in the primary screening, and to analyze the effect of c. 801+2T>G mutation on RhD phenotype by minigene splicing assay. [Methods] The serologic test was performed for RhD phenotype identification and absorption-elution test was performed by using monoclonal anti-D. Sanger sequencing was used to analyze the sequence of RHD genes and the newly identified splicing site mutations of RHD genes were used to construct pSplicePOLR2G micro gene expression plasmids. By using an in vitro micro gene splicing system, the mRNA splicing results were detected and analyzed using agarose and capillary electrophoresis to predict their impact on RhD phenotype. [Results] The serological test results showed that the patient's blood type was RhD-negative, but the anti-D absorption-elution test was positive, indicating a Del phenotype. The rare genotype RHD*(1227A/801+2G) was identified in this individual. The c. 801+2T>G was a novel mutation at 5'-splice site of intron 5. The minigene splicing assay showed that c. 801+2T>G resulted in a complete skipping of RHD exon 5 in the mature transcript, forming a transcript without exon 5. [Conclusion] An individual carrying a novel mutation c. 801+2T>G in the RHD gene was found to exhibit a Del phenotype, but also carry the Asian Del allele c. 1227G>A. It was speculated that the c. 801+2T>G mutation caused RhD negative or Del phenotype based on the results of minigene splicing assay in vitro.
Authors and Affiliations
Shuangshuang JIA, Jizhi WEN, Ling WEI, Runqing ZHANG, Guangping LUO, Yanli JI
Keywords
Related Articles
- EP ID EP754462
- DOI 10.13303/j.cjbt.issn.1004-549x.2024.12.015
- Views 5
- Downloads 0
Expert consensus on RhD blood group testing strategies in blood transfusion compatibility testing laboratories
RhD antigen is the most immunogenic antigens in the Rh blood group system and can cause hemolytic transfusion reactions (HTRs) and severe hemolytic disease of the fetus and newborn (HDFN). RhD-negative red blood cell (RB...
Current situation of blood donation of ethnic minorities in Pu′er City, 2019~2021.
Objective To investigate the participation of ethnic minorities in voluntary blood donation, so as to provide research basis for understanding and promoting blood donation of local ethnic minorities, and further improve...
Identification and pedigree investigation of B(A) and cisAB blood groups
Objective To investigate the serological and molecular genetic characteristics of B(A) and cisAB blood groups discovered in our laboratory. Methods ABO blood group serology and genetic tests were used to identify blood g...
Analysis on testing ability of blood testing laboratories in domestic blood establishments across China
Objective To analyze testing ability of blood testing laboratories in domestic blood establishments, and to comprehensively understand the resource allocation, workload and unqualified blood samples. Methods All blood te...
Analyse the risk factors for producing anti-HLA antibodies in patients with hematological diseases
Objective To explore the risk factors for the production of anti-HLA antibodies in patients with hematological diseases before hematopoietic stemcell transplantation. Methods The results and clinical data of 1 008 patien...