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

ABSTRACT

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