Antibody isotyping for discrimination of disease stage and diagnosis of early Lyme disease.

摘要 每年进行300多万次检测以支持人类莱姆病的实验室诊断 疾病(LD)。而疾控中心常规标准两层(CSTT)方法用于血吸虫病血清学诊断 LD在按照建议使用时运行得相对较好，有很大的改进空间。 与CSTT的补充免疫印迹相关的一些弱点中 值得注意的是，由于对结果的主观视觉解释，重复性较低。要克服 美国疾病控制与预防中心最近更新了基于修改后的STT(MSTT)的建议 因为第二个EIA可以取代免疫印迹。这个项目的主要目标是开发一种 客观、定量、多重EIA可检测四种抗体亚型(IgM/D/G/A)和所有四种抗体 免疫球蛋白亚类(IgG1/2/3/4)，以利用同时获得的抗体概况信息 多个伯氏杆菌抗原建立一种可区分莱姆病分期的方法 提高了总体敏感度，而不会导致特异性的丧失。这项研究的新颖性依赖于： 1)对伯氏杆菌抗原特异性抗体亚型和免疫球蛋白亚类的评价 与莱姆病分期相关；2)开发新的机器诊断工具 学习技术，训练和整合所有数据，并产生客观结果，以便及早区分 由早期播散性/晚期莱姆病引起的莱姆。我们预计这一阶段的SBIR将允许我们开发 一种新的莱姆病血清诊断EIA(IsoEIAplex-LD)及其正在进行的合作 通过DCN诊断使我们的生物标记物适应新的快速侧向血流分析(参见 支持函)，以进行后续第二阶段SBIR。

ABSTRACT More than 3 million tests are performed each year to support the laboratory diagnosis of human Lyme disease (LD). While the CDC conventional standard two-tier (CSTT) approach for serodiagnosis of LD has worked relatively well when used as recommended, there is plenty of room for improvement. Of a number of weaknesses associated with the supplemental immunoblot of the CSTT the most significant is low reproducibility due to the subjective visual interpretation of results. To overcome these weaknesses the CDC recently updated its recommendations based on a modified STT (MSTT) in that a second EIA can replace the immunoblot. The major goal of this project is to develop an objective, quantitative, multiplex EIA that can detect four antibody isotypes (IgM/D/G/A) and all four IgG subclasses (IgG1/2/3/4) to leverage acquisition of simultaneous antibody profile information on multiple B. burgdorferi antigens to build an assay that can discriminate Lyme disease stage with increased overall sensitivity without incurring in loss of specificity. The novelty of this study relies on: 1) evaluation of B. burgdorferi antigen-specific antibody isotypes and IgG subclasses that can be correlated with Lyme disease stage; and 2) development of new diagnostic tools using machine learning techniques to train and integrate all data and produce an objective result to discriminate early Lyme from early disseminated/late Lyme disease. We expect this Phase I SBIR to allow us to develop a new EIA for serodiagnosis of Lyme disease (isoEIAplex-Ld) and to further an ongoing collaboration with DCN diagnostics for the adaptation of our biomarkers to a new rapid Lateral Flow Assay (see Letter of Support) for a follow up Phase II SBIR .