Genetic testing analysis of samples with multiple RhD negative confirmations and inconsistent serological results from voluntary blood donors in Jinan region from 2012 to 2022
Objective:To explore the reasons of repeatedly inconsistent RhD negative serological confirmation results in voluntary blood donors using molecular biology method.Methods:From January 2012 to December 2022,the samples of voluntary blood donors who were initially screened negative for RhD were confirmed by mi-crocolumn agglutination,and RhC,c,E and e were identified and subtyped.Repeatedly inconsistent RhD negative confirmation samples were selected for RhD alleles typing by fluorescent PCR-SSP method.For the specimens whose genotyping could not be confirmed,10 exons of RHD gene were specifically amplified and their genotypes were identified by two directional Sanger sequencing.Results:12 samples in total with repeatedly inconsistent re-sults of the negative confirmation were found to contain RhC or RhE antigens.Five gene types were detected through genetic testing of PCR-SSP and Sanger sequencing,including 8 cases of RHD*01EL.01/RHD*01N.01,1 case of RHD*01N.03/RHD*01N.01,1 case of RHD*15/RHD*01N.01,1 case of RHD*01N.01/RHD*01N.01 and 1 case of RHD*01W.24/RHD*01N.01.Conclusion:Repeatedly inconsis-tent RhD negative confirmation results were found using serological method.Serological and molecular biology method were combined to identify the RhD of voluntary blood donors to ensure the accuracy of blood type identifi-cation and ensure the safety of blood transfusion.
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