Multi Locus High Resolution Typing of SLA Class II Genes, SLA-DQA, -DQB1 and -DRB1, using Genomic DNA Multiplex PCR and Direct Sequencing

We previously reported high resolution typing results of SLA-DQA, DQB1 and DRB1 using genomic PCR and direct sequencing. In this update, we improved our previous protocols and developed multiplex PCR for the simultaneous amplification of those threeSLA class II genes. Then, the PCR results were subjected to separate direct sequencing for SLA-DQA, DQB1 and DRB1 with specific primers for each locus. Using this method, we typed 30 selected individuals comprising 26 class II haplotypes. The results were completely identical to the results of individual locus typing. In addition, this new method helps to overcome the co-amplification of pseudogenes from some samples in the previous DRB1 typing. Our new method can be widely used in the field of SLA typing and contribute to the increase of our knowledge on polymorphisms and functions of SLA molecules.