Jalali Seyedeh Farzaneh, Oodi Arezoo, Azarkeivan Azita, Gudarzi Samira, Amirizadeh Naser
Blood Transfusion Research Center, High Institute for Research and Education in Transfusion Medicine, Hemmat Exp. Way, Next To the Milad Tower, Tehran, P.O. Box: 14665-1157, Iran.
Indian J Hematol Blood Transfus. 2020 Jul;36(3):550-555. doi: 10.1007/s12288-020-01283-y. Epub 2020 Apr 29.
We aimed to determine the JK genotype in thalassemia patients from Iran using different molecular methods to compare with phenotyping results. We also aimed to standardize for the first time, the Tetra-Primer ARMS PCR method for JK genotyping. The serology method cannot correctly determine the phenotype of blood group antigens in patients with multiple blood transfusions. Peripheral blood samples were taken from two hundred alloimmunized thalassemic patients in Tehran Adult Thalassemic Clinic. The samples were tested phenotypically by routine serological methods. After DNA Extraction, SSP-PCR was performed. DNA sequencing and PCR-RFLP were used to confirm the SSP-PCR results. Discrepancies were found between the phenotype and genotype in 32 out of 200 cases. In 16 cases phenotype was determined as Jk (a + b +) but genotype was , in 14 cases phenotype was Jk (a + b +) while the genotype showed , 1 case had been phenotyped as Jk (a + b -) but it was genotyped as and 1 case had been phenotyped as Jk (a - b +) but it was genotyped as . Serological results for a few samples could not be confirmed because of mix-field agglutination. The genotyping however verified the presence of Kidd alleles. Molecular methods are a valuable tool to predict blood group phenotypes in multi-transfused patients in order to select RBC units for a perfect matching improving blood transfusion and preventing alloimmunization. Also Tetra-Primer ARMS PCR is simple and cost effective methods that could be alternative by conventional Molecular methods.
我们旨在使用不同的分子方法确定来自伊朗的地中海贫血患者的JK基因型,以便与表型结果进行比较。我们还旨在首次标准化用于JK基因分型的四引物扩增受阻突变系统聚合酶链反应(Tetra-Primer ARMS PCR)方法。血清学方法无法正确确定多次输血患者血型抗原的表型。从德黑兰成人地中海贫血诊所的200例同种免疫的地中海贫血患者中采集外周血样本。通过常规血清学方法对样本进行表型检测。DNA提取后,进行序列特异性引物聚合酶链反应(SSP-PCR)。使用DNA测序和聚合酶链反应-限制性片段长度多态性分析(PCR-RFLP)来确认SSP-PCR结果。在200例病例中,有32例发现表型与基因型存在差异。16例的表型被确定为Jk(a+b+),但基因型为 ;14例的表型为Jk(a+b+),而基因型显示为 ;1例的表型为Jk(a+b-),但基因分型为 ;1例的表型为Jk(a-b+),但基因分型为 。由于混合视野凝集,一些样本的血清学结果无法得到证实。然而,基因分型证实了基德等位基因的存在。分子方法是预测多次输血患者血型表型的宝贵工具,以便选择红细胞单位进行完美匹配,改善输血并预防同种免疫。此外,四引物扩增受阻突变系统聚合酶链反应是一种简单且经济高效的方法,可以替代传统的分子方法。