Detecting released peptidoglycan fragments as a biomarker for direct diagnosis of acute and chronic Lyme disease

抽象的。 莱姆病是由伯氏疏螺旋体和几个密切相关的基因种引起的。完毕 在过去的25年里，莱姆病病例的数量增加了约1500%，成为第一大疾病 据报道，在美国有媒介传播的疾病。一旦感染，患者会有轻微的流感样症状 可能会变得更严重。在后期，即使在抗生素治疗之后，患者也可以体验到心脏，关节， 以及神经问题。尽管出现了戏剧性的增长，并可能造成毁灭性的后果， 诊断莱姆病仍然是一个重大挑战。目前莱姆病的诊断测试是间接的- 测量对最常见表位的抗体反应(S)。关于暴露的间接检测报告，不活跃 感染。此外，产生可检测的滴度可能需要数周时间，即使如此，假阴性率也是 估计是很高的。这一提议的首席调查者发现，伯氏杆菌大约45%的 肽聚糖(PG)--细菌细胞壁的基本成分--从细胞内进入细胞环境。 在莱姆关节炎患者口服和/或治疗几个月后的滑液中可检出伯氏杆菌PG 静脉注射抗生素。几乎所有的细菌都具有PG，但事实证明，伯氏杆菌的PG极其 独一无二，但在莱姆疏螺旋毛虫中保存。这些发现使我们假设检测到的脱落物 伯氏杆菌PG片段可以作为该病急性和晚期的生物标志物。vbl.使用 以前开发的方法，结合一种新的免疫-PCR方法，我们将测试敏感性 在实验感染的小鼠和人类样本中，在不同的阶段，我们的检测方法的特异性 疾病和治疗。我们的研究承诺为莱姆病提供第一个直接诊断测试。

ABSTRACT. Lyme disease is caused by the bacterium Borrelia burgdorferi, and several closely related genospecies. Over the past 25 years, the number of Lyme disease cases have increased ~ 1500% making it the number one reported vector-borne disease in the United States. Upon infection, patients have mild, flu-like symptoms that can go on to be more severe. In later stages, even after antibiotic therapy, patients can experience heart, joint, and neurological problems. Despite the dramatic increase, and potentially devastating consequences, diagnosing Lyme disease is still a major challenge. Current diagnostic testing for Lyme disease is indirect— measuring an antibody response to the most common epitope(s). Indirect testing reports on exposure, not active infection. Furthermore, producing detectable titers can take weeks, and even still, the false negative rate is estimated to be high. The lead investigator of this proposal discovered that B. burgdorferi sheds ~45% of its peptidoglycan (PG)—the essential component of the bacterial cell-wall—from inside the cell, into its environment. B. burgdorferi PG can be detected in the synovial fluid of Lyme arthritis patients’ months after oral and/or intravenous antibiotics. Virtually all bacteria possess PG, but, as it turns out, the PG of B. burgdorferi is extremely unique, but conserved among Lyme Borreliae. These findings led us to hypothesize that the detection of shed B. burgdorferi PG fragments can act as a biomarker for both acute and late stages of the disease. Using previously developed methods, in conjunction with a novel Immuno-PCR approach, we will test the sensitivity and specificity of our assays, in experimentally infected mice and in human samples, at different stages of disease and treatment. Our studies promise to provide the first direct diagnostic test for Lyme disease.