Novel DNA Microarray for HLA Typing

描述（由申请人提供）：骨髓移植是越来越多疾病的首选临床治疗方法。骨髓的捐赠者和接受者是基于一组复杂的细胞表面蛋白质来匹配的，这组蛋白质被称为HLA（人类白细胞抗原）系统。虽然HLA I类A和B以及HLA II类DRB 1被认为是匹配的关键主要抗原，但该系统是高度多态性的，其中大量等位基因指导每个遗传基因座处的个体多态性。尽管DNA方法目前为成功的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。 在初步研究中，我们开发了一种新的HLA-DNA微阵列方法，该方法有可能为临床HLA分型的高准确性，高效，实用和低成本的检测系统形成新的技术基础。该I期提案被设计为概念验证测试，以确定HLA-DNA微阵列是否可以实现当前DNA方法的准确性和再现性，如应用于II类DRB 1基因座的高分辨率分型。在前期工作中开发的原型HLA-DNA微阵列将通过优化荧光信号发生系统来改进效率，准确性和灵敏度。设计和开发用于制作HLA-DNA微阵列以进行II类DRB 1基因座的高分辨率（等位基因水平）分型的试剂系统和方法。高分辨率IIDRB 1类微阵列将在盲法试验中使用来自不同种族个体的一组200个特征化样本进行评估，并将结果与通过当前DNA方法获得的结果进行比较。如果成功，第一阶段将证明HLA-DNA微阵列技术是合理的，可以满足一个新的，临床上重要的测试平台的要求，并为第二阶段全面开发完整的HLA I类/II类HLA-DNA微阵列分型系统奠定基础。HLA微阵列产品将在支持骨髓和器官移植计划的临床实验室中立即具有商业用途。

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.