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- W2079985742 abstract "Minor histocompatibility antigens (mHA) induce T cell mediated immune responses that have been associated with increased risk of graft versus host disease and allograft rejection. Unlike HLA genes, mHA are encoded by genetically and functionally unrelated genes located throughout the chromosome. The role of mHA in stem cell transplantation and the frequencies of mHA alleles in large study populations remain unknown due in part to the lack of a suitable high throughput method for mHA genotyping. Here we describe the development and utility of a single tube, expandable, multiplexed assay for genotyping mHA (HA-1, HA-2, HA-3, HA-8, HB-1, CD31125, and CD31563) using the Luminex platform. The assay utilizes a multiplexed allele-independent gated amplification of mHA genes followed by differential detection of allele-specific primer products using the MultiCode system (Eragen Biosciences, Madison, WI). The alleles are interrogated using a multiplex allele-specific primer extension reaction with primers tagged with EraCodes, the products hybridized to Luminex beads, and the hybridization duplex detected using streptavidin-PE. Using this assay, we generated an estimate of mHA allele frequencies in a limited local donor population and are assessing the influence of mHA disparities on outcomes of stem cell transplantation in a large cohort of unrelated recipient-donor pairs. Minor histocompatibility antigens (mHA) induce T cell mediated immune responses that have been associated with increased risk of graft versus host disease and allograft rejection. Unlike HLA genes, mHA are encoded by genetically and functionally unrelated genes located throughout the chromosome. The role of mHA in stem cell transplantation and the frequencies of mHA alleles in large study populations remain unknown due in part to the lack of a suitable high throughput method for mHA genotyping. Here we describe the development and utility of a single tube, expandable, multiplexed assay for genotyping mHA (HA-1, HA-2, HA-3, HA-8, HB-1, CD31125, and CD31563) using the Luminex platform. The assay utilizes a multiplexed allele-independent gated amplification of mHA genes followed by differential detection of allele-specific primer products using the MultiCode system (Eragen Biosciences, Madison, WI). The alleles are interrogated using a multiplex allele-specific primer extension reaction with primers tagged with EraCodes, the products hybridized to Luminex beads, and the hybridization duplex detected using streptavidin-PE. Using this assay, we generated an estimate of mHA allele frequencies in a limited local donor population and are assessing the influence of mHA disparities on outcomes of stem cell transplantation in a large cohort of unrelated recipient-donor pairs." @default.
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- W2079985742 date "2005-02-01" @default.
- W2079985742 modified "2023-09-25" @default.
- W2079985742 title "Multiplexed genotyping of human minor histocompatibility antigens" @default.
- W2079985742 doi "https://doi.org/10.1016/j.bbmt.2004.12.156" @default.
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