SBIR-STTR Award

The objective of this research is the development of a HLA (human leukocyte antigens) typing method at DNA level using oligonucleotide array technology. The HLA genes are the most polymorphic gene in human with over 550 known alleles in HLA-A, -B, - C and -DR and -DQ loci. Characterization of the HLA genes at the allele level is crucial for successful human organ/marrow transplantation. Here we describe an advanced HLA typing method for one-step typing of the three major HLA loci, HLA-A, -B and -DR at intermediate to high allelic resolution level. Our long term goal is to c:evelop a fully automated HLA typing system for the six major HLA loci (A, B, C, DR, DQ and DP). This technology allows a high resolution genotyping with tens of thousands different oligonucleotide probes per assay through a single hybridization reaction. In phase I period, we will design initial oligonucleotide arrays, develop a sample preparation method, and evaluate the sensitivity and specificity of the method. The HLA typing using this technology is expected to overcome limitations faced by current DNA-based HLA typing methods such as resolution, robustness, and speed. PROPOSED COMMERCIAL APPLICATIONS: A DNA-level tissue typing method capable of a one step complete typing of the major Class I and II HLA alleles using oligonucleotide arrays should provide a valuable tool in a large scale screening as well as a routine rapid HLA typing of organ/marrow donors and recipients. Revenues will be derived from the sales of the system.

The objective of this proposal is to develop a commercial HLA (human leucocyte antigen) typing system using a microsphere-based DNA probe array. Characterization of HLA gene at DNA level has become a crucial tool in identifying HLA compatible donor-recipient pairs for successful human organ/marrow transplantation. Here, we describe an advanced yet tangible method of HLA typing for the five major HLA loci, HLA-A, B, C, DRB, and DQB to accommodate the needs of clinical laboratories. The microsphere-based array platform has a unique combined advantage of DNA probe array, and flexibility, cost-effectiveness, and sample processing capability of flow cytometry. Multi-analyte DNA hybridization assay using a panel of HLA locus-specific probe is conducted and analyzed in a single reaction. In phase II period, we will continue and expand probe development for both low and high resolution typing of the five major HLA loci, develop sample amplification method, develop data analysis software, and rigorously validate the final system through beta-testing. Flexibility, cost-effectiveness, and automation potential of the proposed system is expected fulfill the existing needs of HLA typing at DNA level for laboratories involved in solid organ transplant as well as high throughput screening for the national bone marrow donor registry.