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）：莱姆病的准确，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，由于伯格多菲尔芽孢杆菌分离株的高遗传异质性，即使是从单个位置收集的疾病，该疾病的血清诊断尤其困难。此外，目前针对莱姆病的血清诊断测试缺乏敏感性和特异性，无法在该疾病的早期阶段检测B. burgdorferi感染。为了解决这些缺陷，我们将基于多个疏螺旋体属的基因种子来制造蛋白质微阵列。用于开发灵敏的单层莱姆病测定法。使用基因组和蛋白质组学方法，我们将首先进行疏螺旋体基因组之间的全面比较，以识别B. burgdorferi sensu stricto的独特，膜相关和/或正面选择的抗原。然后，我们将开发由这些单独的Burgdorferi蛋白质组成的蛋白质组微阵列，并使用来自B. burgdorferi感染的个体的血清介绍了体液免疫反应。我们假设，渗透菌株的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巨大差异。我们使用B. burgdorferi菌株B31，N40，JD1和297的蛋白质数据的初步蛋白阵列数据已经表明，我们的测定法的敏感性远远超过了两种疾病早期患者的两种商业可用的莱姆病测定法。因此，使用微阵列对疏螺旋体蛋白质组的全面研究将确定在感染过程中表达的抗原蛋白的光谱。我们将利用这些信息扩展我们的阵列平台，以开发一种敏感和特定的单层莱姆病测定法，该测定可能具有针对莱姆病的广泛活性和特异性。该技术最终可以改编成在美国，欧洲和亚洲发现的其他毛毛虫物种，并用于开发诊断性的莱姆病测试以用于全球使用，最终可能有可能包括晚期症状的预后概率。 公共卫生相关性：由于莱姆病很难被初级保健医生诊断，因此我们将开发一种新的诊断测试，能够在其发作时检测到该疾病。通过全面识别患者认识的莱姆蛋白，我们将能够识别所有患者在第一次来医生办公室时对所有患者做出反应的关键蛋白质。这最终将允许开发办公测试，这将有助于与治疗相关的快速启动治疗。