Diagnostic Assays for early Lyme Borreliosis using in-vivo expressed antigens

描述（由申请人提供）：莱姆病是由硬蜱传播的螺旋体伯氏疏螺旋体感染引起的，是美国最常见的病媒传播疾病，2009年向CDC报告了38，000多例确诊和疑似病例。螺旋体首先在皮肤中建立感染，然后传播到内部器官，如心脏，关节和神经系统，在那里可以看到疾病。感染在早期发现时对抗生素的反应最大，但那些患有播散性感染或治疗延迟的人可能会经历对抗生素无反应的衰弱性疾病。莱姆病的及时准确诊断对于优化治疗反应和预防疾病的长期后遗症至关重要。检测抗B抗体的血清学试验（ELISA和免疫印迹）。目前，莱姆病的最敏感和广泛可用的实验室测试。这些试验利用培养的B实验室菌株的全细胞裂解物。作为抗原来源的伯氏螺旋体，因此不能检测到B的抗体。Burgdorferi蛋白主要在体内表达。推荐的两层方法（通过免疫印迹确认阳性或可疑ELISA）在感染早期仅具有约30%的灵敏度，并且由于B的交叉反应性，特异性降低。伯氏抗原与其他传染性病原体的抗原。该I期申请旨在通过使用体内表达的B来改进目前可用的莱姆病血清学试验。作为抗原的伯氏特异性蛋白质。我们最近发现了一组新的B。在B的前几周中表达的伯氏螺旋体基因和蛋白质。莱姆病早期患者的血清在IgG ELISA上对其有反应。我们建议使用来自明确定义的莱姆病病例的人血清样本（未暴露于B的受试者），在多重格式检测中评估这些候选抗原的灵敏度和特异性。burgdorferi和其它感染性和风湿性疾病。肽针阵列将以ELISA形式绘制候选抗原与莱姆病血清的免疫显性表位，以并入精细的多重测定中。这项I期研究的结果将为开发新的敏感和特异性莱姆病血清学检测奠定基础，这些检测更准确地反映B。在感染宿主中激发免疫应答的伯氏抗原。 公共卫生相关性：莱姆病，由于感染螺旋体伯氏疏螺旋体，可导致多系统疾病，涉及皮肤，心脏，关节和神经系统。目前莱姆病的血液检测在感染早期对抗生素最敏感时并不敏感。这项应用将确定一种基于螺旋体最初感染皮肤时表达的一组新蛋白质的莱姆病新检测方法是否能准确地检测出莱姆病的早期阶段。

DESCRIPTION (provided by applicant): Lyme disease, due to infection with the Ixodes tick-transmitted spirochete Borrelia burgdorferi, is the most common vector-borne disease in the United States, with over 38,000 confirmed and probable cases reported to the CDC in 2009. Spirochetes first establish infection in the skin and then disseminate to internal organs such as the heart, the joints, and the nervous system where disease can be seen. The infection is most responsive to antibiotics when identified early, but those with disseminated infection or in whom treatment is delayed can experience debilitating disease that can become unresponsive to antibiotics. The timely and accurate diagnosis of Lyme disease is essential for optimizing the response to therapy and for preventing long- term sequelae of the disease. Serologic tests (ELISA and immunoblot) that detect antibodies against B. burgdorferi are currently the most sensitive and widely available laboratory tests for Lyme disease. These tests utilize whole-cell lysates of cultured laboratory strains of B. burgdorferi as a source of antigens, and therefore do not detect antibodies to B. burgdorferi proteins expressed predominantly in vivo. The recommended two-tiered approach in which a positive or equivocal ELISA is confirmed by immunoblot has only ~30% sensitivity in early stages of infection, and specificity is reduced due to cross-reactivity of B. burgdorferi antigens with those of other infectious agents. This Phase I application seeks to improve upon the currently available serologic tests for Lyme disease through the use of in vivo expressed, B. burgdorferi- specific proteins as antigens. We have recently identified a novel panel of B. burgdorferi genes and proteins that are expressed in the first few weeks of B. burgdorferi infection and to which sera from patients with early Lyme disease react on IgG ELISA. We propose to evaluate these candidate antigens in multiplex format assay for their sensitivity and specificity using human sera samples from well-defined cases of Lyme disease, subjects who have had no known exposure to B. burgdorferi, and other infectious and rheumatic diseases. Peptide pin arrays will map the immunodominant epitopes of candidate antigens with Lyme disease sera in an ELISA format for incorporation into a refined multiplex assay. The results of this Phase I study will set the stage for the development of new sensitive and specific serologic tests for Lyme disease that more accurately reflect the B. burgdorferi antigens inciting immune responses in the infected host. PUBLIC HEALTH RELEVANCE: Lyme disease, due to infection with the spirochete Borrelia burgdorferi, can cause a multisystem illness involving the skin, heart, joints and nervous system. Current blood tests for Lyme disease are not as sensitive early in infection when the infection is most responsive to antibiotics. This application will determine whether a new assay for Lyme disease based on a novel panel of proteins expressed by spirochetes when they initially infect the skin accurately detects Lyme disease in its earliest stages.