中国人群中Del表型分子背景研究  

Study on the molecular background of Del phenotype in Chinese population

作　　者：李勤[1] 叶璐夷[2] 郭忠慧[2] 钱旻[1] 朱自严[2] 

机构地区：[1]华东师范大学生命科学学院,上海上200062 [2]海市血液中心血型参比实验室

出　　处：《中华医学遗传学杂志》2006年第5期486-491,共6页Chinese Journal of Medical Genetics

基　　金：国家自然科学基金(30371312)~~

摘　　要：目的研究Del表型的分子基础,并探寻新的Del等位基因。方法 515名经血清学方法确定的RhD阴性血样用吸收放散试验筛选并确认Del表型。Del的基因组DNA通过多重PCR(multiplex poly-merase chain reaction,MPX PCR)检测RHD基因;用序列特异性引物PCR(sequence-specific primer-polymerase chainreaction,PCR-SSP)检测Del等位基因:RHD(K409K)和RHD(M295I)。经PCR-SSP筛选,结果为阴性的样品通过基因组DNA测序及cDNA测序分析其分子背景。结果吸收放散试验共检出Del 79名。经多重PCR检测,79份样品均带有RHD第3、4、5、6、7、9特异性外显子。PCR-SSP结果显示,75名Del具有RHD(K409K)等位基因,无Del样品带有RHD(M295I)等位基因。经测序发现4名新RHD等位基因:RHD3G→A(GenBank DQ310735) ,RHD28C→T,RHD53T→C(GenBank DQ451877、DQ451878) ,RHD251T→C(GenBank DQ310734)。结论 Rh血型系统非常复杂,新的D变异表型和新的RHD等位基因可能被不断发现。Objective To elucidate the molecular background of Del phenotype in the Chinese population and explore new Del alleles. Methods Five hundred and fifteen RhD negative blood samples was tested by Rh typing test, indirect antiglobulin test and adsorption and elution assay to screen the Del phenotype, DNA of all the Del samples was analysed by multiplex polymerase chain reaction （MPX PCR） for the presence of RHD and by sequence-specific primer polymerase chain reaction （PCR-SSP） for Del alleles： RHD 1227A and RHD 885T. Samples which showed the negative result by PCR-SSP, were additionally analysed by genomic DNA sequencing and cDNA sequencing. Results Seventynine Del samples were found by adsorption and elution assay. All these samples bad RHD exons 3, 4, 5, 6, 7 and 9. Except 4 Del samples, other 75 Del samples carried the RHD 1227A allele. None of the samples bad the RHD 885T allele. Four novel RHD alleles were found in these four Del sample. There were RHD 3G→A （GenBank DQ310735）, RHD 28C→T, RHD 53T→C （GenBank DQ451877,DQ451878）, RHD 251T→C （GenBank DQ310734）. Conclusion Rh blood group system is very complex. New D variation phenotypes and new RHD alleles may be discovered ceaselessly.

关 键 词：RHD基因 DEL表型 遗传多态性 等位基因 测序 

分 类 号：R394[医药卫生—医学遗传学]