Novel DNA Microarray for HLA Typing

描述（申请人提供）：骨髓移植是越来越多疾病的首选临床治疗方法。骨髓供体和受体的匹配是基于一组复杂的细胞表面蛋白，称为HLA（人类白细胞抗原）系统。虽然HLA I类A和B以及HLA II类DRB1被认为是对匹配至关重要的主要抗原，但该系统具有高度多态性，在每个遗传位点上都有大量等位基因指导个体多态性。虽然DNA方法目前为成功移植提供了最佳水平的匹配，但这些方法成本高，劳动密集，并且在提供优化移植结果所需的HLA等位基因的精细分辨率方面能力有限。

DESCRIPTION (provided by applicant): Bone marrow transplantation is a preferred clinical treatment for an expanding number of diseases. Donors and recipients of bone marrow are matched based on a complex group of cell surface proteins called the HLA (Human Leukocyte Antigen) system. While HLA class I A and B and HLA class II DRB1 are believed to be major antigens critical for matching, the system is highly polymorphic with a large number of alleles directing individual polymorphisms at each genetic locus. Although DNA methods currently provide the best level of matching for successful transplants, these methods are costly, labor intensive and limited in their ability to provide the refined resolution of HLA alleles necessary to optimize transplant outcome. In preliminary studies, we developed a new HLA-DNA Microarray methodology which has the potential to form a new technology base for a highly accurate, yet, efficient, practical and low cost assay system for clinical HLA typing. This Phase I proposal is designed as a proof-of-concept test to determine whether HLA- DNA Microarrays can achieve the accuracy and reproducibility of current DNA-methods as applied to high resolution typing of the class II DRB1 locus. The prototype HLA-DNA Microarray developed in preliminary work will be modified to improve efficiency, accuracy and sensitivity by optimizing the fluorescence signal generating system. The reagent system and methods for fabricating HLA-DNA Microarrays to perform high resolution (allelic level) typing of class II DRB1 loci will be designed and developed. High resolution class IIDRB1 Microarrays will be evaluated in a blinded trial using a set of 200 characterized samples from ethnically diverse individuals and the results compared with those obtained by current DNA methods. If successful, Phase I will demonstrate that the HLA-DNA Microarray technology is sound and can fulfill the requirements of a new, clinically important testing platform and set the stage for full-scale development of the complete HLA class I/class II HLA-DNA Microarray typing system in Phase II. The HLA-Microarray product will have immediate commercial utility in clinical laboratories supporting bone marrow and organ transplantation programs.