HLA typing methods used in clinical laboratories utilize either sequence-specific primers (SSP) to amplify or sequence certain HLA alleles, or sequence-specific oligonucleotide probes (SSOP) to detect them. However, control DNA standards for primer- and probe-based HLA typing systems are not available commercially, and market research among HLA typing laboratories indicates a desire for kits to monitor primer- and probe- based HLA typing reagents. Phase I studies indicate the feasibility of producing HLA DNA standards that consist of modified gene sequences cloned in a plasmid vector. The authors predict that the constructs will be useful as controls for both SSP-PCR and SSOP assays.
The specific aims of this Phase II proposal are: 1) to develop and test additional HLA DNA standards, including those for low resolution HLA-A, -B, and DQB primers and probes, 2) to optimize this set of low-resolution HLA DNA standards, and 3) to evaluate the feasibility of streamlining the standards for future kit development.
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