Development of PCR-ELISA Technique for Determination of HLADRB1*01 Group Alleles

Document Type: Research Paper

Authors

1 Department of Medical Biotechnology, Faculty of Medical Science, Tarbiat Modarres University, Tehran, I.R. Iran.

2 Department of Hematology, Faculty of Medical Science, Tarbiat Modarres University, Tehran, I.R. Iran.

3 Department of Medical Biochemistry, Faculty of Medical Science, Tarbiat Modarres University, Tehran, I.R. Iran.

Abstract

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.

Keywords