Rapid identification of minor histocompatibility antigen HA-1 subtypes H and R using fluorescence-labeled oligonucleotides

• Published in: Tissue antigens 2000-11, Vol.56 (5), p.449-452
• Main Authors: Kreiter, S., Wehler, T., Landt, O., Huber, C., Derigs, H.-G., Heß, G.
• Format: Article
• Language: English
• Subjects: allogeneic transplantation; Fluorescent Dyes; GvHD; HA-1; HA-1 antigen; histocompatibility antigen HLA; Humans; hybridization probes; LightCycler instrument; Minor Histocompatibility Antigens - classification; Minor Histocompatibility Antigens - genetics; Oligonucleotides; Oligopeptides - classification; Oligopeptides - genetics; PCR; Time Factors
• ISSN: 0001-2815; 1399-0039
• DOI: 10.1034/j.1399-0039.2000.560509.x

Acknowledgments: We thank Brigitte Schuch and Karola Schmidt for excellent technical assistance. This work was supported by a grant of the Deutsche Krebshilfe (Nr. 70‐2427 and Nr. 70‐2428). : Donor‐recipient disparitiy of the minor histocompatibility antigen HA‐1 is relevant for the development of graft‐versus‐host disease after HLA‐matched sibling allogeneic bone marrow transplantation in HLA‐A*0201‐positive individuals. Two different alleles of HA‐1 with a single amino acid polymorphism have been identified. Here we describe a time‐ and cost‐efficient method for HA‐1 typing of genomic DNA, using site‐specific hybridization probes with the LightCycler. This method was compared with standard techniques as sequencing or allele‐specific polymerase chain reaction (PCR) and proved to be specific, reliable and reproducible. We conclude that HA–1‐subtyping using fluorescent‐labeled oligonucleotides represents a attractive method for the screening of samples before allogeneic transplantation in HLA‐A*0201‐positive individuals.