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: 8138668
• 负责人: BENJAMIN J LUFT
• 金额: $ 23.48万
• 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 2013-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8138668
• 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）：准确、可靠的莱姆病诊断分析是确保成功治疗和完全康复的关键。然而，由于伯氏疏螺旋体分离株的高度遗传异质性，即使在从单一地点收集的分离株中，该疾病的血清诊断也特别困难。此外，目前的莱姆病血清诊断检测缺乏在疾病早期检测伯氏疏螺旋体感染的敏感性和特异性。为了解决这些不足，我们将制作基于伯氏疏螺旋体多基因种的蛋白质微阵列，用于开发敏感和特异性的单层莱姆病检测。利用基因组学和蛋白质组学方法，我们将首先对伯氏疏螺旋体基因组进行全面比较，以鉴定独特的、膜相关的和/或正选择的伯氏疏螺旋体严格感抗原。然后，我们将开发由这些个体伯氏疏螺旋体蛋白组成的蛋白质组微阵列，并使用伯氏疏螺旋体感染个体的血清来描述体液免疫反应。我们假设伯氏疏螺旋体菌株基因含量的差异将解释以B31分离物作为测试参考的人血清蛋白阵列中抗原反应的巨大差异。我们使用伯氏疏螺旋体菌株B31、N40、JD1和297蛋白的初步蛋白质阵列数据已经显示，我们的检测方法在疾病早期患者中的敏感性远远超过两种市售的莱姆病检测方法。因此，利用微阵列对伯氏疏螺旋体蛋白质组进行全面的研究将确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，开发一种敏感和特异性的单层莱姆病检测，该检测可能具有广泛的莱姆病活性和特异性。这项技术最终可用于包括在美国、欧洲和亚洲发现的其他伯氏疏螺旋体，并用于开发一种世界范围内使用的莱姆病诊断测试，最终可能有可能包括晚期症状的预后概率。