Application of Allele Specific Oligonucleotide (ASO) PCR in determination of Rh antigens in thalassemic patients

Thalassemia is a genetic disease whit relative or complete lack of alpha or beta globin chain. Patients with moderate and severe form need to have multitransfusion in early life. Occurrence of all immunization against blood group antigens in patients with thalassemia is relatively high and may have difficulties in treatment and transfusion. Antibody production against this blood groups cause lots of problems like preparation of compatible blood for transfusion. Correct diagnosis of blood group phenotype due to existence of dual population of donor and patient RBC would be difficult. In this study, randomly from 40 thalassemic patients, before blood transfusion, 4cc of peripheral blood collected in tubes containing EDTA. Also, 10 healthy individuals who had no history of blood transfusion were in the study as control for serological (agglutination) and molecular results. Phenotyping of patients and control group was done by tube agglutination method by commercial antibody. For molecular test, Allele Specific Primer Amplification (ASPA) PCR was performed for each antigens in separate micro tubes. In this study' we could set up a method that can amlify any of Rh C c E and E gene separately by a pair of specific primer in a same thermal conditions.Comparison of results of two methods showed that in the control group with no transfusion history. Similar results in phenotyping and genotyping was observed. But in patients, results of two methods had lots of differences. Discussion and The advantage of this method over PCR-RFLP method is that' all four genes can be amplified the in a same concentration and temperature conditions, and ability to determine the individual's antigen, immediately by electrophoresis. Therefore, Since the above method is simple and inexpensive for medical and research centers it is recommended.