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.

对于标准化的血清诊断莱姆病测试而存在着巨大的公共卫生需求，该测试将感染与由于外表面蛋白A（OSPA）疫苗接种引起的假阳性区分开。将使用缺乏FDA最近批准的莱姆病疫苗的主要成分OSPA抗原的Borrelia burgdorferi抗原制剂开发新的酶联免疫吸附测定法（ELISA）。疫苗接收者产生的OSPA抗体通常会引起血清阳性测试结果，无论是否发生实际感染。常规的莱姆病ELISA使用包括表达的OSPA在内的B. bugdorferi的全细胞裂解物。将以ELISA格式评估三种不同的OSPA阴性抗原候选候选制剂 - 变异菌株选择，质粒基因破坏和重组抗原表达 - 将以ELISA格式进行评估。敏感性和特异性将使用代表莱姆病，正常人血清和其他感染性或自身免疫性疾病的患者的早期和晚期的血清板确定。最佳性能ELISA将在商业参考实验室环境中进行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将取代基于标准B. burgdorferi菌株的全细胞裂解物的当前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。