Serological and molecular biological analysis of a patient with RHD-CE(3-7)-D gene recombination and RHCE*cE variant
Objective To investgate a case of RHD-CE(3-7)-D gene recombination and RHCE*cE variant for serological and molecular biological characteristics,to provide experimental basis for the clinical safe blood transfusion.Methods ABO,RhD,and RhCcEe antigens were identified using the micro-column gel card method.Blood group was determined using the blood test tube method,and irregular antibodies were screened using the anti-human globulin card method.The genotypes of RhD,RhC,Rhc,RhE,and Rhe were detected using PCR-SSP,and the entire sequence of the RHD/RHCE gene was analyzed using three-genera-tion full-length sequencing.Results ABO,RhD,and RhCcEe blood group antigens were tested as follows:A(-),B(-),RhD(1+),RhC(4+),Rhc(1+),RhE(1+),Rhe(4+)using a micro-column gel card.Upon repeating the test using the test tube method,the results were A(-),B(-),RhD(w+),RhC(4+),RhC(w+),RhE(w+),RhE(4+).Anti-human globulin card screening for irregular antibody was negative.Geno-typing results of RhDCcEe by PCR-SSP:RHD(+),RHC(+),RHc(+),RHE(+),RHe(+).Analysis of the RHD/RHCE gene showed that RHD haploid 1 had a complete deletion in exons 1-10,while haploid 2 exons showed a RHD-CE gene recombination,it was confirmed that its recombination type was RHD-CE(3-7)-D.The starting point of this recombination was identified in exon 2(g.20238-20312),and the end point was deter-mined in exon 8(g.49184-50480).Additionally,a new base point mutation was found in exon 6 of the RHCE gene(827C>A).Conclusion RHD-CE(3-7)-D gene recombination and RHCE*cE(827C>A)novel allele mutation may cause weak expression of D and CE blood group antigens.This provides a foundation for safe blood transfusions in clinical settings.
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