浙江汉族DEL表型基因分型方法的建立

为了快速鉴定DEL表型，建立浙江汉族DEL表型的基因分型方法，根据RHD1227A等位基因序列设计等位基因特异性．聚合酶链反应（allele specific-polymerase chain reaction，AS-PCR），用于DEL表型的基因分型。用已知RHD1227A多态性的样本和已知DEL表型的样本评价基因分型方法的灵敏度和特异性。结果表明：在50例RHD1227A阳性和50例RHD1227A阴性的Rh阴性样本中基因分型方法的灵敏度和特异性都是100％；在33例DEL阳性样本和89例DEL阴性的样本中，基因分型方法的灵敏度为100％，有2例样本血清学结果为阴性而基因分型结果为阳性，重新用血清学方法和序列分析方法复核这2例样本，发现2例都是血清学漏检，因而基因分型方法的特异性是100％。结论：设计的基因分型方法可以有效地鉴定浙江汉族Rh阴性人群中的DEE表型。

Establishment of Genotyping Method for DEL Phenotype in Zhejiang Har Population

In order to establish a genotyping method for DEL phenotype in Zhejiang Han population, an AS-PCR method was developed according to the RHD 1227A allele sequence. Its specificity and sensitivity were assessed in two Rh negative populations whose RHD 1227A or DEL phenotype status was known. The results showed that in evaluation of the method by detecting 50 RHD 1227A positive and 50 RHD 1227A negative individuals, the genotyping method displayed a sensitivity of 100% and a specificity of 100%; in evaluation of the method by detecting 33 DEL positive and 89 DEL negative individuals, the sensitivity was 100%, however, there were two serologically negative samples which were confirmed as positive using genotyping method. After re-testing these two samples with serological method and sequence analysis, it was found that original serological method gave false negative results and genotyping method still showed 100% specificity. The minimal target DNA concentration of this genotyping method is 8.13 ng/microl. In conclusion, designed genotyping method can be used to identify DEL phenotype efficiently in Zhejiang Han Rh negative population.