A Novel single-tier Lyme disease assay using Borrelia burgdorferi Protein Arrays.

使用伯氏疏螺旋体蛋白阵列进行新型单层莱姆病测定。

• 批准号: 8324939
• 负责人: BENJAMIN J LUFT
• 金额: $ 24.32万
• 依托单位: STATE UNIVERSITY NEW YORK STONY BROOK
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-09-01 至 2014-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8324939
• 关键词: Novel; single; tier; Lyme; disease

DESCRIPTION (provided by the applicant): Accurate, reliable diagnostic assays for Lyme disease are critical to ensure successful treatment and full recovery. However, serodiagnosis of the disease is particularly difficult due to the high level of genetic heterogeneity of B. burgdorferi isolates, even among those collected from a single location. Furthermore, current serodiagnostic tests for Lyme disease lack sensitivity and specificity for detecting B. burgdorferi infection in the early stages of the disease. To address these deficiencies, we will fabricate protein microarrays based on multiple genospecies of Borrelia spp. for use in developing a sensitive and specific single-tier Lyme disease assay. Using genomic and proteomic approaches, we will first perform comprehensive comparisons between Borrelia genomes to identify unique, membrane associated and/or positively selected antigens of B. burgdorferi sensu stricto. We will then develop proteome microarrays consisting of these individual B. burgdorferi proteins and profile the humoral immune response using sera from B. burgdorferi infected individuals. We hypothesize that differences in the gene content of Borrelia strains will account for the dramatic differences in the antigenic response of human sera in protein arrays that used the B31 isolate as test reference. Our preliminary protein array data using proteins from B. burgdorferi strains B31, N40, JD1 and 297 has already revealed that the sensitivity of our assay far exceeds that of the two commercially-available Lyme disease assays in patients in the early stages of the disease. Thus, a comprehensive investigation of the Borrelia proteome using microarrays will identify the spectrum of antigenic proteins expressed during infection. We will utilize this information to expand upon our array platform to develop a sensitive and specific single-tier Lyme disease assay that will potentially have a wide range of activity and specificity against Lyme disease. This technology can ultimately be adapted to include other Borrelia species found in the USA, Europe and Asia, and used to develop a diagnostic Lyme disease test for world-wide use that may eventually have the potential to include prognostic probability of late symptoms. PUBLIC HEALTH RELEVANCE: Since Lyme disease is a difficult infection to diagnose by the primary care physician, we will develop a new diagnostic test that is able to detect the disease at its onset. By comprehensively identifying the Lyme proteins that the patient recognizes, we will be able to identify the key proteins which all patients respond to when they first come to the doctor's office. This will ultimately allow the development of an office test which will facilitate the quick initiation of therapy which is associated with cure.

描述（由申请人提供）：莱姆病准确、可靠的诊断测定对于确保成功治疗和完全康复至关重要。然而，由于伯氏疏螺旋体分离株（即使是从单一地点收集的分离株）也具有高度的遗传异质性，因此该疾病的血清诊断特别困难。此外，目前莱姆病的血清诊断测试缺乏检测疾病早期阶段的伯氏疏螺旋体感染的敏感性和特异性。为了解决这些缺陷，我们将制造基于疏螺旋体属多个基因种的蛋白质微阵列。用于开发敏感且特异的单层莱姆病检测。使用基因组和蛋白质组学方法，我们将首先在疏螺旋体基因组之间进行全面比较，以鉴定严格伯氏疏螺旋体的独特的、膜相关的和/或阳性选择的抗原。然后，我们将开发由这些单独的伯氏疏螺旋体蛋白质组成的蛋白质组微阵列，并使用来自伯氏疏螺旋体感染个体的血清来分析体液免疫反应。我们假设疏螺旋体菌株基因含量的差异将解释使用 B31 分离株作为测试参考的蛋白质阵列中人血清抗原反应的显着差异。我们使用伯氏疏螺旋体菌株 B31、N40、JD1 和 297 的蛋白质进行的初步蛋白质阵列数据已经表明，我们的检测方法的灵敏度远远超过两种市售的莱姆病早期患者检测方法的灵敏度。因此，使用微阵列对疏螺旋体蛋白质组进行全面研究将鉴定感染期间表达的抗原蛋白谱。我们将利用这些信息扩展我们的阵列平台，开发一种敏感且特异的单层莱姆病检测方法，该检测方法可能对莱姆病具有广泛的活性和特异性。这项技术最终可以适用于在美国、欧洲和亚洲发现的其他疏螺旋体属物种，并用于开发供全世界使用的莱姆病诊断测试，最终有可能包括晚期症状的预后概率。 公共卫生相关性：由于莱姆病是一种初级保健医生难以诊断的感染，我们将开发一种新的诊断测试，能够在疾病发作时检测到该疾病。通过全面识别患者识别的莱姆病蛋白，我们将能够识别所有患者第一次来医生办公室时都会产生反应的关键蛋白。这最终将允许开发办公室测试，这将有助于快速启动与治愈相关的治疗。