Abstract： AIM:To develop a pyrosequencing-based typing (PSBT) approach for high resolution identification of HLA-DRB alleles. METHODS:The DNA fragments from HLA-DRB exon-2 were obtained using allele-specific or multiplex PCR amplification. The PSBT of purified DNA was performed. RESULTS:The polymorphic residues of HLA-DRB genes were identified in each pyrosequencing reaction and read length up to 90 nucleotides was obtained. A blood sample containing heterozygous alleles (DRB1-0405 and DRB3-01011) was analyzed using purified DNA respectively from allele-specific or multiplex PCR reaction, the results of which were consistent with those from the traditional method. CONCLUSION:Pyrosequencing used in the analysis of HLA-DRB alleles has the advantage of high resolution and can be widely employed clinically in donor/recipient selection.