[Molecular blood genotyping in patients with Thalassemia major in Tehran adult thalassemic clinic]

Blood typing by serologic methods after transfusion has limitations due to presence of donor red cells in recipients. Accurate determination of red blood cells [RBCs] antigens is very important in multitransfused patients including beta-thalassemics and sickle cell anemics. So, the aim of this study was to evaluate DNA- based methods as supplement to the hemagglutination technique to determine the red blood cell [RBC] antigen profile of multitransfused patients with beta- thalassemia. DNA was extracted from peripheral blood of 20 apparently normal people and 44 patients including 35 with beta- thalassemia [out of whom 19 had clinical evidence of delayed hemolytic transfusion reaction], 8 with thalassemia intermedia [out of whom 2 had hemolytic reaction], and one with sickle cell thalassemia. RHD/ RHC/ RHc/ RHE/ RHe/ JKA/ JKB/ FYA/ FYB/ KELL1/ KELL2 alleles were determined by PCR and were then compared with the hemagglutination method. Phenotype and genotype results were the same in all controls. The phenotypes and genotypes of 53 blood antigens of 26 patients were incompatible. Most of the discrepancies [19 cases] occurred in the Rh system, and fifteen in the Duffy and Kidd systems. The results show that screening platelet concentrates for bacterial contamination is necessary for blood transfusion centers and hospital blood banks. Blood typing by serologic method was not accurate in this study but genotyping could determine true blood groups in multitransfused patients and help in selection of RBCs without alloimmunized antigens in future transfusion attempts. Specificity, sensitivity, positive and negative predictive values of hemagglutination method for RhD antigen had good values in comparison to the molecular method. This might be due to pre- transfusion determination of RhD for thalassemic patients so as to receive Rh- matched blood units. It seems pre-transfusion blood typing of Rh and Kell antigens, which are the cause of hemolytic reactions, in comparison to the molecular method could be cost effective. In addition, typing of Rh and Kell antigens in some regular blood donors could be helpdul for selecting antigen-negative RBCs for transfusion dependent patients

Mini-pool screening for HCV infection in Iranian blood donors: preliminary results

Screening the blood donors for serological markers reduced the incidence of transfusion transmitted infections especially post-transfusion hepatitis C. However, there remains residual risk due to pre-seroconversion period. HCV RNA [PCR] of blood donations reduced theresidual risk of transfusion-transmitted HCV infection. In this study, blood donations werescreened for HCV RNA by RT-PCR method.An extra plasma sample was collected from 1026 blood donors. 1000 out of 1026 sampleswere negative for HBsAg, anti-HCV [EIA, third generation], anti-HIV and RPR. Every 5samples were pooled. The sensitivity of HCV-RNA detection by RT-PCR method was 380geq/ml according to Proficiency VQC panel. 1000 donations in 200 pools were tested.False reactivity of samples considered positive accounts for 5.5% of cases, and 5.5% were invalid due to non-specilic bands. 6% of the pools were false-positive. A false positive result was defined as positive on initial testing but negative on repeat single testing. However, all ofthe samples were negative for HCV RNA by RT-PCR method.No sample was found to be serologically negative and HCV RNA positive. However, further studies are recommended for further clarification

HLA-Class I and II frequencies in ethnic Hamedani people, and the comparison of serological and molecular [PCR] methods for HLA-Class II

In this study our aim was to determine HLA-Class I and II antigens freguencies of Hamedani ethnic group. In addition to demographic studies and disease association, it has wide application in bone marrow donor registeries. In order to establish DNA-based HLA typing in central Laboratory of Iranian Blood Transfusion Organization, a comparison of serological and molecular [sequence specific primers "SSP"] methods for HLA-DRB has been performed. The study was descriptive and the population under study were selected out of the native people of Hamedan; 100 healthy volunteer blood donors were chosen by questionnaire. 10ml heparinized and 3ml EDTA blood were collected from each selected donor. EDTA [PCR] samples were then frozen.N.I.H standard microlymphocytotoxicity and Nylon wool T and B cells separation was used for serological I and II typing. HLA-Class I plates were prepared from Iranian Blood Fractionation and Research Company and for Class II we used Biotest DR/DQ Typing Trays. PCR was done using "Roche high pure DNA extraction" Kit and HLA-DRBSSP [Biotest]. The most and least frequent HLA-B antigens were B5 group [B51/B52] and B16 [38,39] respectively. Because of low resolution of HLA-DRB Kit, no significant difference was observed between serological and PCR methods. Although some blanks have been determined by PCR.The HLA-DRB determination by PCR is mandatory for donor/recipient pairs [even sibling] for bone marrow transplantation; for donors it should be done by high resolution kits