Development of PCR-ELISA technique for determination of HLA DRB1*01 group alleles

We have developed an allotyping assay for detection of four HLA DRB1*01 group alleles based on polymerase chain reaction and solution hybridization in a microtiter plate. Using group specific primers a region within exon 2 of HLA DRB1 gene was amplified by PCR. Labeling of PCR product was achieved by adding small amount of Dig-dUTP in place of dTTP. Labeled PCR product was hybridized to allele (HLA DRB1*0101, *0102, *0103 and *0104) specific and a group (HLA DRB1*01) specific oligonucleotide probes in separate wells of the plate. Hybridized amplicones were detected by an enzymatic procedure. Ninety DNA samples were tested in parallel with PCR-SSP typing. The results were found to be well correlated by two methods. These results further suggest that, PCR-ELISA would be a rapid, specific and simple method that can be used for high resolution HLA typing before bone marrow and stem cell transplantation.