RHD genotype and zygosity analysis in the Chinese Southern Han D+, D− and D variant donors using the multiplex ligation‐dependent probe amplification assay

• Published in: Vox sanguinis 2017-10, Vol.112 (7), p.660-670
• Main Authors: Ji, Y. L., Luo, H., Wen, J. Z., Haer‐Wigman, L., Veldhuisen, B., Wei, L., Wang, Z., Ligthart, P., Lodén‐van Straaten, M., Fu, Y. S., Schoot, C. E., Luo, G. P.
• Format: Article
• Language: English
• Subjects: Alleles; Amplification; Antigens; Asian Continental Ancestry Group - genetics; Assaying; Bioassays; Blood & organ donations; Blood cells; Blood Donors; Deoxyribonucleic acid; DNA; DNA probes; DNA sequencing; Erythrocytes; Exons; Genotype; Genotype & phenotype; Genotyping; Humans; Molecular Diagnostic Techniques - methods; Molecular Diagnostic Techniques - standards; multiplex ligation‐dependent probe amplification; Multiplex Polymerase Chain Reaction - methods; Multiplex Polymerase Chain Reaction - standards; Multiplexing; Phenotype; Population genetics; Rh-Hr Blood-Group System - genetics; RHD variant alleles; Sampling methods; the Chinese population; Zygosity
• ISSN: 0042-9007; 1423-0410
• DOI: 10.1111/vox.12554

Background and Objectives Several comprehensive genotyping platforms for determining red blood cell (RBC) antigens have been established and validated for use in the Caucasian and Black populations, but not for the Chinese. The multiplex ligation‐dependent probe amplification (MLPA) assay was validated for RHD genotyping in the Chinese. Materials and Methods The blood samples of 200 D+, 200 D− and 62 D variant Chinese donors were collected. RhD antigen was routinely typed by serological method. D variant phenotype was determined by an anti‐D panel (D‐Screen), when RBCs were available. The RHD genotype and its zygosity were analysed with the RH‐MLPA technique. When the MLPA was unable to identify a RHD variant, direct sequencing of all exons of the RHD gene was performed. Results In 200 D+ donors, DD (168/200, 84%), D (12/200, 6%), DDD genotype (1/200) and D variant allele carriers (19/200, 9·5%) were found. In 200 D− donors, six reported RHD alleles, RHD*01EL.01, RHD*01N.03, RHD*01N.05, RHD*01N.16, RHD*DFR2 and RHD*weak partial 15 and one novel RHD*1154T allele were identified in 36·5% (73/200) of them. In 62 D variant donors, three novel RHD alleles, RHD*79_81delCTC, RHD*710T and RHD*689A, and twelve reported alleles, RHD*DVI.3, RHD*weak partial 15, RHD*DVI.4, RHD*01EL.01, RHD*01N.03, RHD*DLO, RHD*DV.5, RHD*D‐CE(2‐10), RHD*730C, RHD*weak D type 25, 33 and 72, were identified, either alone or in combination. Conclusion The RH‐MLPA assay correctly identified the common RHD variant alleles in the Chinese population. However, DNA sequencing was required to identify certain alleles; probes to detect these alleles should be added into the assay.