Informative immunodiagnostics for Lyme disease

描述(由申请人提供)：长期目标是提供一种改进的免疫分析作为莱姆病诊断的辅助手段。该检测最好具有(A)在疾病早期检测感染的更高的敏感性，(B)对于疾病的所有阶段，如果不是比目前可用的检测方法更好的话，那么它具有良好的特异性，以及(C)具有足够的信息量，以使化验解释人员推断患者感染的伯氏杆菌菌株。此R21/R33应用的具体目的如下：(1)使用全基因组蛋白质组阵列来询问LD患者和感染伯氏杆菌的实验动物的IgM反应。这种方法将类似于成功用于研究人类和小鼠对这种病原体的一系列免疫球蛋白反应的方法。通过纳入两种或更多的SPC类型来提高IgM检测的灵敏度的假设将得到检验。(R21阶段)(2)调查对亚单位抗原的抗体反应性图谱是否为推断某人感染的伯氏杆菌菌株提供了附加值。这将在感染了不同菌株的实验动物中进行评估，并使用从感染菌株中恢复的患者的血清进行评估。假设(A)OspC类型特异性反应可以区分，以及(B)对BBK07和BBK12蛋白的反应模式随感染菌株的不同而不同，将进行测试。(R21阶段)(3)开发一种用于最终临床测试的免疫分析方法，方法是：(A)选择信息量最大的一组重组抗原，以获得较高的预测值，同时将成本降至最低；以及(B)暂时从ELISA、免疫试条和Luminex中选择最合适的平台，用于在分析的临床研究评估中实施。抗原的选择将从先前研究中确定的抗原以及AIMS 1和2的结果中选择(R33阶段)。 与公共卫生相关：该项目的长期目标是改进对莱姆病病原体伯氏疏螺旋体的免疫反应的实验室测试。这将通过在感染的早期阶段提高抗体检测的灵敏度，并通过提供患者可能感染的病原体菌株的证据来实现。

DESCRIPTION (provided by applicant): The long-term goal is to provide for an improved immunoassay as an adjunct for the diagnosis of Lyme disease. The assay preferably would have (a) greater sensitivity for detecting infection during early disease, (b) have as a good if no better specificity than currently available assays, alone or in combination, for all stages of disease, and (c) be sufficiently informative that assay interpreters infer the strain of B. burgdorferi a patient is infected with. Specific aims for this R21/R33 application are the following: (1) Use a genome-wide proteome array to interrogate IgM responses of patients with LD and experimental animals infected with B. burgdorferi. The approach will be similar to what was successfully used for studies with an array of IgG responses of humans and mice to this pathogen. The hypothesis that sensitivity of IgM assays is heightened by including two or more OspC type will be tested. (R21 phase) (2) Investigate whether profiles of antibody reactivity to sub-unit antigens provide added-value for inference about the strain of B. burgdorferi someone has been infected with. This will be evaluated in experimental animals infected with different strains and with sera from patients for whom the infecting strain was recovered. Hypotheses that (a) OspC type-specific responses can be distinguished and (b) patterns of reactivity to the BBK07 and BBK12 proteins vary with the infecting strain will be tested. (R21 phase) (3) Develop an immunoassay for eventual clinical testing by (a) selecting the most informative set of recombinant antigens for achieving a high predictive value while minimizing cost, and (b) selecting the most suitable platform, provisionally from among ELISA, immunostrips, and Luminex, for implementation in clinical research evaluation of the assay. The choice of antigens will be from those identified in a previous study, as well as from the results of aims 1 and 2. (R33 phase) PUBLIC HEALTH RELEVANCE: The long-term goal of the project is improvement in laboratory testing for immune responses to the Lyme disease agent, Borrelia burgdorferi. This will be achieved by increasing the sensitivity of assays for antibodies during the early stage of the infection and by providing evidence about the strain of the pathogen a patient may be infected with.