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PCR-SSP RHD基因定型方法的初步应用

Application of a PCR-SSP method for RHD genotyping

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【作者】 周一炎秦建江

【Author】 ZHOU Yi-yan1, QIN Jian-jiang2(1.Department of Shenzhen Blood Center, Shenzhen, Guangdong 518035, P.R.China; 2.Department of Hami Regional Blood Bank, Xinjiang 830000, P.R.China)

【机构】 深圳市血液中心新疆哈密地区中心血站 广东深圳518035新疆830000

【摘要】 目的采用序列特异性引物PC R技术(PC R-SSP)进行R H D基因定型。方法采用新近报道的PC R-SSP R H D定型方法检测10例R hD阳性个体,包括2例哈萨克族人、2例蒙古族人、2例藏族人和4例维吾尔族人,以及11例R hD阴性个体(汉族10例、维吾尔族人1例),并通过测序和内含子检测分析1例汉族个体的R H D基因型。结果全部样品的PC R-SSP检测结果与血清学表型相符,1例汉族R hD阴性个体鉴定为D放散型,携带R H D1227A等位基因,1例个体的检测结果提示携带融合R H D基因。测序分析显示该融合基因的第1、2和第10外显子序列与正常R H D基因相应外显子相同,而缺失其余外显子,由于R H D基因和R H C E(e)基因的第2外显子序列相同,笔者采用PC R方法检测R H D基因第2内含子,结果为阴性,说明该个体携带的融合基因为R H D-C E(2-9)-D2等位基因。结论PC R-SSP可用于中国人R h血型D抗原基因定型。

【Abstract】 RHD genotyping through previously reported sequence specific primer-polymerase chain reaction (PCR-SSP) method. Rh D phenotype was predicted and genotyped through a PCR-SSP method in 10 Rh-positive samples from 2 kazakh, 2 Mongol, 2 Tibetan and 4 uigur, and 11 Rh-negative sample from 10 Hans and 1 uigur. RHD sequencing and intron detection were applied in one sample to determine hybrid RHD allele. The PCR-SSP results of all samples are consistent with serological phenotypes. One of Rh-negative Hans was determined as Del, carrying RHD1227A allele, and one was indicated carrying hybrid RHD. Sequencing analysis of this sample showed that exon 1, 2 and 10 have identical sequences as normal RHD while others was tested negative. As RHD and RHCE(e) has same sequence in exon 2, RHD exon 2 with PCR in the sample were detected. The result was negative, which indicated this individual carried RHD-CE(2-9)-D2 allele. [Conclusion] The PCR-SSP method is applicable for RHD genotyping in Chinese.

【关键词】 R h血型D抗原R H D基因R H C E基因基因交换
【Key words】 rhesus blood groupD antigenRHDRHCEgene conversion
  • 【文献出处】 中国现代医学杂志 ,China Journal of Modern Medicine , 编辑部邮箱 ,2005年07期
  • 【分类号】Q75
  • 【被引频次】1
  • 【下载频次】72
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