中国浙江义乌人群RhD真阴性和RhDel表型的分子机制研究

本研究探讨中国浙江义乌人群RhD阴性表型的分子机理。在献血人群中用盐水凝集试验筛选RhD阴性样本，并对这些阴性样本进一步通过抗人球蛋白试验和吸收放散技术鉴定真阴性和RhDel表型。然后，采用PCR—SSP法特异性扩增RhD真阴性和Rhdel样本的RHD基因10个外显子，对扩增阳性的外显子分别进行DNA序列分析。结果发现：在38例初筛RhD阴性的样本中，吸收放散法得到30例RhD真阴性和8例RhDel样本（占盐水法阴性样本的21．1％）。在30例RhD真阴性中，28例样本PCR扩增10个外显子均为阴性，2例样本仅外显子1，2和10为阳性；8例RhDel样本的10个外显子扩增均为阳性，测序发现8个样本均存在1227G〉A变异。结论：中国浙江义乌人群RhD真阴性以RHD基因全缺失为主，也存在一定比例的外显子部分缺失，RhDel表型分子机制均为1227G〉A变异。

Molecular Basis for Real RhD Negative and RhDel Phenotypes in Yiwu Population of Zhejiang Province in China

This study was purposed to investigate the molecular basis for RhD negative phenotype in Yiwu population in Zhejiang Province of China. The RhD negative samples were screened by saline agglutination test in blood donors. Some real RhD negative and RhDel phenotypes were identified using anti-human globulin test and absorbtion elution test. Ten exons of RHD gene in these samples were amplified by PCR-SSP, and positive exons were DNA sequenced. The results indicated that 30 real RhD negative and 8 RhDel phenotypes were identified in 38 initial RhD negative samples. Ten exons were complete negative in 28 real RhD negative samples and only exon 1, 2 and 10 were positive in 2 real RhD negative samples amplified by PCR. All 10 exons in 8 RbDel samples were positive and a DNA variant （ 1227G 〉 A） was found in 8 RhDel samples. It is concluded that all exons are absenced in most real RhD negative phenotypes, and the partial exons absence is also found in some real negative phenotypes among Yiwu population in Zhejiang province of China. The G to A mutation at position 1227 is found in all RhDel phenotypes.