Blood group genotyping in alloimmunized multi-transfused thalassemia patients from Iran.

OBJECTIVES: Serological methods may not be reliable for RBC antigen typing, especially in multi-transfused patients. The blood group systems provoking the most severe transfusion reactions are mainly Rh, Kell, Kidd, and Duffy. We intended to determine the genotype of these blood group system antigens among Iranian alloimmunized thalassemia patients using molecular methods and compare the results with serological phenotyping. METHODS: Two hundred patients participated in this study. Blood group phenotype and genotype were determined using the serological method and PCR-SSP, respectively. The genotypes of patients with incompatibility between phenotype and genotype were re-evaluated by RFLP-PCR and confirmed by DNA sequencing. RESULTS: Discrepancies between phenotype and genotype results were found in 132 alleles and 83 (41.5%) patients; however, there was complete accordance between the three genotyping methods. Most discrepancies were detected in Rh and Duffy systems with 47 and 45 cases, respectively, and the main discrepancy was in the FY*B/FY*B allele when serologically showed Fy(a+b+). All 39 undetermined phenotypes, due to mixed-field reactions, were resolved by molecular genotyping. CONCLUSION: Molecular genotyping is more reliable compared with the serological method, especially in multi-transfused patients. Therefore, the addition of blood group genotyping to serological assays can lead to an antigen-matched transfusion in these patients.

Kidd Blood Group Genotyping for Thalassemia Patient in Iran.

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 JK*A/JK*A, in 14 cases phenotype was Jk (a + b +) while the genotype showed JK*B/JK*B, 1 case had been phenotyped as Jk (a + b -) but it was genotyped as JK*A/JK*B and 1 case had been phenotyped as Jk (a - b +) but it was genotyped as JK*A/JK*B. 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.