Document Type : Research Paper
Department of Medical Biotechnology, Faculty of Medical Science, Tarbiat Modarres University, Tehran, I.R. Iran.
Department of Hematology, Faculty of Medical Science, Tarbiat Modarres University, Tehran, I.R. Iran.
Department of Medical Biochemistry, Faculty of Medical Science, Tarbiat Modarres University, Tehran, I.R. Iran.
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, PCRELISA would be a rapid, specific and simple method
that can be used for high resolution HLA typing before bone marrow and stem cell transplantation.