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Molecular and family analyses revealed two novel RHD alleles in a survey of a Chinese RhD‐negative population
Author(s) -
Ye L.Y.,
Guo Z.H.,
Li Q.,
Zhu Z.Y.
Publication year - 2007
Publication title -
vox sanguinis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.68
H-Index - 83
eISSN - 1423-0410
pISSN - 0042-9007
DOI - 10.1111/j.1423-0410.2006.00886.x
Subject(s) - genetics , allele , biology , exon , haplotype , rh blood group system , population , polymerase chain reaction , gene , intron , microbiology and biotechnology , medicine , antibody , environmental health
Background and Objectives RHD alleles are considered variable in the Chinese RhD‐negative persons. The purpose of the present work was to elucidate the molecular bases of two novel RHD alleles identified in a survey of a Chinese RhD‐negative population. Materials and Methods A total of 163 RhD‐negative blood samples were investigated. The sequences of RHD exons were examined by RHD exon specific multiplex polymerase chain reaction (PCR) and PCR with sequence‐specific primers (PCR‐SSP). Characterizations of RHD intron 2 and Rhesus box were performed by PCR‐PstI digestion. The DNA and cDNA sequences of the novel alleles were determined by PCR and reverse transcriptase‐PCR (RT‐PCR) sequencing analysis. A family study was performed to investigate the segregation of a novel RHD allele. Results One hundred and twenty‐nine samples (79·1%) had no RHD gene. Twenty‐seven samples (16·6%) carried RHD‐CE‐D hybrid alleles. The remainder seven samples (4·3%) appeared to have an intact RHD gene. Three of them were sequenced for RHD gene and two novel alleles, RHD 325del and RHD intron 2 1A , were identified. The deletion of a nucleotide A at position 325 in the allele RHD 325del resulted in a stop codon at amino acid position 118. The RHD intron 2 1A allele was generated from a splice mutation and its transcript sequence had no exon 2. Family study indicated that the RHD 325del allele was inherited with a Ce haplotype. Conclusion This study provides the molecular bases of RHD alleles RHD 325del and RHD intron 2 1A . The existences of RHD 711del , RH (D1 CE2‐9 D10) , and RH (D1 CE2‐9 D10) alleles in the Chinese population were confirmed. A PCR‐SSP‐based assay for rapid detection of RHD 325del and RHD intron 2 1A alleles was established and it could be used to predict the RHD genotype in the Chinese RhD‐negative subjects.