Khosroshahi Behzad Nazel, Oodi Arezoo, Namjou Saba, Gholamali Tahereh, Amirizadeh Naser
Blood Transfusion Research Center, High Institute for Research and Education in Transfusion Medicine, Hemmat Highway, Next to the Milad Tower, Tehran, Iran.
Indian J Hematol Blood Transfus. 2019 Jan;35(1):119-124. doi: 10.1007/s12288-018-0992-3. Epub 2018 Aug 1.
D antigen is the most important and immunogenic antigen of the Rh blood group. The RhD-negative phenotype has different genetic backgrounds with variable distribution in different populations. Hybrid , resulting from gene deletion, is used in genotyping studies of the Rh blood group as a marker to identify the gene deletion. This study for the first time identified genetic mechanisms for the occurrence of RhD-negative phenotype among the Iranian population. 200 RhD-negative blood donors were randomly selected from Tehran Blood Transfusion Center. The phenotype of D, C, Ε, e and c antigens was serologically identified, and DNA was extracted from buffy coat. The molecular analysis of hybrid was performed by PCR-SSP and PCR-RFLP. Moreover, the presence of different exons of gene was investigated by real-time PCR on extracted DNA. Hybrid was detected in all samples, and PCR-RFLP confirmed that 198 (99%) were for an gene deletion and 2 were for hybrid in which one (0.5%) had a weak D type 11 and the other one (0.5%) had a --- hybrid allele. Similar to Caucasians, the frequency of gene deletion was high among the Iranian population studied in this investigation, so hybrid can be used as an efficient marker to detect gene deletion in our population.
D抗原是Rh血型系统中最重要且具有免疫原性的抗原。RhD阴性表型具有不同的遗传背景，在不同人群中的分布各异。由基因缺失导致的杂合子在Rh血型系统基因分型研究中被用作识别基因缺失的标记。本研究首次确定了伊朗人群中RhD阴性表型出现的遗传机制。从德黑兰输血中心随机选取200名RhD阴性献血者。通过血清学方法鉴定D、C、E、e和c抗原的表型，并从 Buffy 层中提取DNA。采用聚合酶链反应-序列特异性引物法（PCR-SSP）和聚合酶链反应-限制性片段长度多态性法（PCR-RFLP）对杂合子进行分子分析。此外，通过对提取的DNA进行实时聚合酶链反应研究基因不同外显子的存在情况。在所有样本中均检测到杂合子，PCR-RFLP证实198例（99%）为基因缺失的杂合子，2例为杂合子，其中1例（0.5%）为弱D11型，另1例（0.5%）为---杂合等位基因。与高加索人相似，在本研究的伊朗人群中基因缺失的频率较高，因此杂合子可作为检测我们人群中基因缺失的有效标记。
