番禺地区RHD变异体基因分型研究

目的研究分析番禺地区RHD变异体基因分型特征。方法采用微量板法对2010年11月～2011年8月到本站参加献血的26 172名次献血者进行RhD阴性筛查;采用抗球蛋白法对初筛RhD阴性的标本进行确认,采用吸收放散试验对经抗球蛋白法确认为RhD阴性的标本进行Del表型筛选;采用RH基因变异体基因分型检测试剂盒(PCR-SSP法)对献血者模板DNA进行扩增,对扩增产物进行电泳分析,根据电泳图谱判断RHD基因分型。对血清学与基因分型结果不符的标本进行测序分析。结果初筛检出60名RhD阴性,经抗球蛋白法确认59例为RhD阴性,阴性频率为0.23%;吸收放散试验检出10例Del表型。60例初筛阴性的基因分型检测发现40例RHDEXON1～10全缺失基因型,发生频率约为67.8%;10例RHD 1227A DEL基因型,发生频率约为16.9%;7例部分D,发生频率约为11.9%;1例RHD弱D15基因型,发生频率约为1.2%;2例RHD基因分型阳性需作进一步碱基测序分析。结论本地区血清学RhD阴性献血者中,存在RHD 1227A DEL、RHD-CE(2-9)-D,RHD-CE(5)-D、RHD-CE(6-9)-D和RHD弱D15等基因型,血清学结合基因分型能更准确鉴定RhD血型。

Genotyping study of RHD variants among RhD-negative blood donors, Panyu district

Objective To study and analyze genotyping characteristics of RHD gene variants among Panyu RhD-negative blood donors.Methods Use micro-plate method to screen RhD-negative samples among 26 172 blood donors from November 2010 to August 2011 in Panyu Blood Centre.Using anti-globulin method to identify RhD-negative screening samples,using adsorption and elution test to screen Del phenotype among the above RhD-negative-identified samples,using RH gene variants genotyping assay kit(PCR-SSP) to amplify template DNA of blood donors and the amplified products were analyzed by electrophoresis.And RHD genotyping was determined according to the polyacrylamide gel electrophoresis.The samples that the serological and genotyping results were not matching were sequenced.Results 60 RhD negative samples were screened out among 26 172 blood donors,and 59 samples were confirmed RhD negative by anti-globulin method,with the negative frequency of 0.23%.Adsorption and elution test detected out 10 cases of Del phenotype.Genotyping detected the 60 RhD-negative samples,40 cases of RHD EXON1~10 deletion genotype were found,with the approximately frequency of 67.8%,and 10 cases of RHD 1227A DEL,with the approximately frequency of 16.9%,and 7 cases of partial D,with the approximately frequency of 11.9%,and 1 cases of RHD weak D15 genotype,with the approximately frequency of 1.2%,and 2 cases needed further allele sequencing.Conclusion In Panyu district,among the serological RhD-negative blood donors,RHD 1227A DEL,RHD-(2-9)CE-D,RHD-CE(5)-D,RHD-CE(6-9)-D and RHD weak D15 genotype etc.were existing.Adsorption and elution test combined with PCR-SSP,could further improve the accuracy of RhD blood group identification.