OSPA-NEGATIVE ELISA FOR LYME DISEASE

存在对标准化血清诊断莱姆病测试的显著公共卫生需求，该标准化血清诊断莱姆病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。一种新的酶联免疫吸附试验（ELISA）将使用缺乏OspA抗原的伯氏疏螺旋体抗原制剂开发，OspA抗原是最近FDA批准的莱姆病疫苗的主要成分。疫苗接受者产生OspA抗体，通常导致血清阳性测试结果，无论是否发生实际感染;常规莱姆病ELISA使用B的全细胞裂解物。包括表达的OspA。将以ELISA形式评价在I期使用三种不同方法（变异株选择、质粒基因破坏和重组抗原表达）启动的三种不同OspA阴性抗原候选制剂。将使用代表莱姆病早期和晚期的血清组、正常人血清和来自其他感染性或自身免疫性疾病患者的血清来确定灵敏度和特异性。将在商业参考实验室环境中进一步开发和广泛评价性能最佳的ELISA。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B全细胞裂解物的ELISA。burgdorferi菌株拟议的商业应用：这项研究将导致一个血清学ELISA诊断人类莱姆病用于商业参考实验室。

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.