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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。
Objective To analyze the genotyping characteristics of RHD variants in Panyu area. Methods A total of 26 172 blood donors who participated in the blood donation from November 2010 to August 2011 were screened for RhD-negative by microtiter plate method. RhD negative specimens were identified by antiglobulin method. In the test, Rh phenotype was confirmed by anti-globulin method. The phenotypes were determined by PCR. The template DNA was amplified by PCR-SSP. The amplification products were amplified Electrophoresis analysis, RHD genotyping based on electrophoresis patterns. Serological and genotyping results do not match the samples were sequenced. Results Sixty negative RhDs were detected by primary screening. 59 cases were negative by RhD and the negative frequency was 0.23%. Ten cases of Del phenotypes were detected by absorption and desorption assay. The genotypes of 60 cases with negative primary screening revealed 40 cases of RHDEXON1 ~ 10 complete deletion genotype, the frequency was about 67.8%; 10 cases of RHD 1227A DEL genotype, the frequency was about 16.9%; 7 cases of part D, the frequency of occurrence About 11.9%; 1 case of RHD weak D15 genotype, frequency of about 1.2%; 2 cases of RHD genotypes positive for further base sequencing analysis. Conclusion RHD 1227A DEL, RHD-CE (2-9) -D, RHD-CE (5) -D, RHD-CE (6-9) -D and RHD weak D15 Other genotypes, serological typing and genotyping can more accurately identify RhD blood group.