OSPA-NEGATIVE ELISA FOR LYME DISEASE

莱姆病 OSPA 阴性 ELISA

• 批准号: 6212800
• 负责人: Raymond Allen Koski
• 金额: $ 36.57万
• 依托单位: L2 DIAGNOSTICS, LLC
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-07-01 至 2002-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6212800
• 关键词: Borrelia; Lyme disease; bacterial proteins; communicable disease diagnosis; diagnosis design /evaluation; diagnosis quality /standard; enzyme linked immunosorbent assay; human tissue; recombinant proteins; serology /serodiagnosis; surface antigens

A significant public health need exists for a standardized serodiagnostic Lyme disease test that distinguishes infection from false positives due to outer surface protein A (OspA) vaccination. A new enzyme-linked immunosorbent assay (ELISA) will be developed using Borrelia burgdorferi antigen preparations that lack OspA antigen, the primary constituent of the Lyme disease vaccine recently approved by the FDA. Vaccine recipients generate OspA antibodies typically causing seropositive test results whether or not actual infection occurred; conventional Lyme disease ELISAs use whole-cell lysates of B. burgdorferi including expressed OspA. Three different OspA-negative antigen candidate preparations initiated in Phase I using three different approaches -- variant strain selection, plasmid gene disruption, and recombinant antigen expression -- will be evaluated in ELISA formats. Sensitivity and specificity will be determined using serum panels representing early and late stages of Lyme disease, normal human sera, and sera from patients with other infectious or autoimmune diseases. The best performing ELISA will be further developed and evaluated extensively in a commercial reference laboratory setting. We anticipate that as the OspA vaccine becomes widely used, the new OspA-negative ELISA will supplant current ELISAs based on whole-cell lysates of standard B. burgdorferi strains. PROPOSED COMMERCIAL APPLICATIONS: This research will lead to a serologic ELISA diagnostic for human Lyme disease for use in commercial reference laboratories.

公共卫生领域迫切需要标准化的莱姆病血清诊断测试，以区分感染和外表面蛋白 A (OspA) 疫苗接种引起的假阳性。将使用缺乏 OspA 抗原的伯氏疏螺旋体抗原制剂开发一种新的酶联免疫吸附测定 (ELISA)，OspA 抗原是 FDA 最近批准的莱姆病疫苗的主要成分。疫苗接种者产生的 OspA 抗体通常会导致血清检测结果呈阳性，无论是否发生实际感染；传统的莱姆病 ELISA 使用伯氏疏螺旋体的全细胞裂解物，包括表达的 OspA。第一阶段启动的三种不同的 OspA 阴性抗原候选制剂将使用三种不同的方法（变体菌株选择、质粒基因破坏和重组抗原表达）以 ELISA 形式进行评估。将使用代表莱姆病早期和晚期阶段的血清组、正常人血清以及来自其他传染病或自身免疫性疾病患者的血清来确定敏感性和特异性。性能最佳的 ELISA 将在商业参考实验室环境中得到进一步开发和广泛评估。我们预计，随着 OspA 疫苗的广泛使用，新的 OspA 阴性 ELISA 将取代目前基于标准伯氏疏螺旋体菌株全细胞裂解物的 ELISA。拟议的商业应用：这项研究将导致人类莱姆病的血清学 ELISA 诊断，用于商业参考实验室。