PCR-SSP RHD基因定型方法的初步应用

目的采用序列特异性引物PCR技术(PCR-SSP)进行RHD基因定型.方法采用新近报道的PCR-SSP RHD定型方法检测10例RhD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RhD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RHD基因型.结果全部样品的PCR-SSP检测结果与血清学表型相符,1例汉族RhD阴性个体鉴定为D放散型,携带RHD1227A等位基因,1例个体的检测结果提示携带融合RHD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RHD基因相应外显子相同,而缺失其余外显子,由于RHD基因和RHCE(e)基因的第2外显子序列相同,笔者采用PCR方法检测RHD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RHD-CE(2-9)-D2等位基因.结论 PCR-SSP可用于中国人Rh血型D抗原基因定型.

Application of a PCR-SSP method for RHD genotyping

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.