Borrelia antigen detection assay for urine-based diagnosis of early Lyme disease

 描述（由申请人提供）：血清学检测仍然是最广泛使用的实验室工具，以支持莱姆病（LB）的诊断，俗称“莱姆病”。“这种方法在某些临床应用中效果很好，但在其他方面有很大的局限性。针对致病细菌伯氏疏螺旋体的特异性IgM和/或IgG抗体应答可能在感染后持续数十年，尽管进行了有效治疗。因此，血清学检测不能区分活动性和既往LB，也不能作为治疗后症状持续的患者的“治愈测试”。然而，血清学检测最著名的局限性是其在早期感染中的敏感性差。尽管LB的诊断和治疗的重要性，在其最早（局部）阶段，以防止传播和随后的发展严重的，有时是永久性的病理后来在感染的过程中，血清学检测是假阴性的70-80%的患者目前与局部游走性红斑（EM），皮疹的早期LB。当典型的“靶心”EM皮疹出现在适当的流行病学背景下时，很容易做出诊断，但这仅代表10-20%的EM病例。更典型地，EM病变可能难以与由其他感染性和非感染性疾病引起的皮疹区分开;此外，在大约20%的早期LB病例中完全不存在皮疹。 值得注意的是，上述限制可能都可以通过可以直接检测感染原而不是检测病原体的测试来解决。 宿主对病原体免疫反应我们建议通过发现和验证新的B，为开发一种新的和高度准确的早期LB测试奠定基础。早期LB患者尿液中的伯氏菌蛋白生物标志物。使用已被证明的质谱（MS）方法，以前应用于发现新的诊断生物标志物的活动性结核病（TB）和内脏利什曼病（VL）的研究人员，我们的团队已经确定了一个有前途的候选人B。来自确诊EM患者的尿样中的伯氏螺旋体生物标志物。我们现在建议在这一有希望的初步发现的基础上，对来自确诊EM的成人和儿童患者的另外20份尿样进行MS，使我们能够选择最佳生物标志物候选物，用于使用我们在TB和VL生物标志物工作中审查的方法开发抗原检测测定。然后，我们将通过检测来自患有和未患有确诊早期LB的成人和儿童患者的大量尿液样本来验证生物标志物候选物的诊断效用。我们预计这项工作将提供坚实的概念证明，B。因此，可以在早期LB患者的尿液中检测到伯氏螺旋体蛋白抗原，从而为随后的测定优化和大规模验证研究奠定了坚实的基础。我们预计，我们的合作将导致莱姆诊断领域的实质性进展。

 DESCRIPTION (provided by applicant): Serologic testing remains the most widely-used laboratory tool to support the diagnosis of Lyme borreliosis (LB), colloquially known as "Lyme disease." This approach works well for some clinical applications but has major limitations in others. A specific IgM and/or IgG antibody response against the causative bacterium, Borrelia burgdorferi, may persist for decades after infection despite effective treatment. Therefore, serologic testing cannot differentiate between active and past LB, and cannot be used as a "test of cure" in patients whose symptoms persist despite therapy. The most well-known limitation of serologic testing, however, is its poor sensitivity in early infection. Despite the importance of diagnosis and treatment of LB in its earliest (localized) phase to prevent dissemination and subsequent development of severe and sometimes permanent pathology later in the course of infection, serologic testing is falsely negative in 70-80% of patients who present with localized erythema migrans (EM), the rash of early LB. When a classic "bull's eye" EM rash presents in appropriate epidemiologic context, the diagnosis is readily made, but this represents only 10-20% of EM cases. More typically, EM lesions can be difficult to distinguish from rashes caused by other infectious and non-infectious diseases; moreover, a skin rash is completely absent in approximately 20% of early LB cases. Notably, the limitations described above could potentially all be addressed by a test that can directly detect the infectious agent, rather than detecting the host's immune response to that agent. We propose to lay the groundwork for development of a novel and highly accurate test for early LB by discovering and validating novel B. burgdorferi protein biomarkers in the urine of patients with early LB. Using proven mass spectrometric (MS) methods previously applied to the discovery of novel diagnostic biomarkers of active tuberculosis (TB) and visceral leishmaniasis (VL) by the investigators, our team has identified a promising candidate B. burgdorferi biomarker in urine samples from patients with confirmed EM. We now propose to build on this promising preliminary finding by performing MS on an additional 20 urine samples from adult and pediatric patients with confirmed EM, allowing us to select the top biomarker candidates for development of antigen detection assays using methods vetted in our TB and VL biomarker work. We will then validate the diagnostic utility of the biomarker candidates by testing a large panel of urine samples from adult and pediatric patients with and without confirmed early LB. We anticipate that this work will provide firm proof of concept that B. burgdorferi protein antigens can be detected in the urine of patients with early LB, thus laying a solid foundation upon which to build in subsequent assay optimization and large-scale validation studies. We anticipate that our collaboration will lead to a substantial advance in the Lyme diagnostics field.