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[Serological and Molecular Biological Analysis of 56 Cases with Weak D Antigen Expression and 14 Cases with Anti-D
Hong-Mei Yang1,2, Si-Fei Ma1,2, Xin Zou1,2
1Changzhou Blood Center, Changzhou 213000, Jiangsu Province, China.
Objective:
To explore the serological weak expression of the RhD antigen in 56 samples and the serological and molecular biological characteristics of 14 samples with anti-D pattern detected by the RhD negative confirmation test, and to investigate the correlation between haplotype genotypes and RhCcEe phenotypes for accurate identification of anti-D and anti-LW.
Methods:
A total of 56 samples with serologically confirmed weak RhD antigen expression were selected from 169 538 blood donors, and haplotype detection was conducted using PacBio SMRT third-generation sequencing technology. For the 14 serologically confirmed RhD negative samples exhibiting anti-D pattern, absorption-elution tests, SSP-PCR (c.1227G>A), differential tests for anti-D and anti-LW antibodies, and haplotype detection were performed.
Results:
Ten known alleles (51/56) and 5 novel alleles (5/56) were identified. The known alleles included RHD*weak partial 15(20/56), RHD*DVI.3(14/56), RHD*weak D type33 (5/56), RHD*weak partial 15/RHD*01EL.01(4/56), RHD*DCS2 (3/56), RHD*weak partial 15/RHD*01N.04(1/56), RHD*weak D type38 (1/56), RHD*weak D type17(1/56), RHD*DV.5(1/56), RHD*DFR1(1/56). The novel alleles consisted of RHD*01N.04 with c.781C>G, RHD*01EL.01 with.c.487-2A>G, RHD*01N.01 with c.781C>G, RHD*weak D33 with c.953G>C, RHD*01N.01 with RHD-CE(10) recombination. The RhCcEe phenotypes for the 14 anti-D pattern samples were ccee (7/14), Ccee (2/14), CCee (2/14), CcEe (1/14), ccEe (1/14), and ccEE (1/14); the alleles were RHD*01N.01(7/14) and RHD*01 EL.01(7/14).
Conclusion:
Through serological and molecular biology analysis, the RhD blood type was accurately detected, multiple alleles were identified. The RhCcEe phenotype and allele profiles of 14 samples with anti-D patterns were clarified, which helps to correctly identify anti-D and anti-LW.
