Direct visualization assay for detection of B. burgdorferi in early Lyme Disease

描述（由申请人提供）：莱姆病是一种由螺旋体伯氏疏螺旋体引起的蜱传感染，在美国每年估计有30万例。目前批准的免疫反应测试是次优的，通常在疾病早期给出阴性结果。本申请中概述的研究是开发一种直接可视化B的诊断血液测试。burgdorferi。这种测试将允许更早的检测-从而更好的治疗-并可用于监测有症状的患者对治疗的反应。RareCyte Inc.，该公司是一家早期生命科学公司，致力于鉴定和回收血液中的稀有细胞，拥有基于试管、浮子和收集机制的高度工程化富集技术，可以按密度对血液中的细胞和病原体进行分层。使用该技术，已经开发出原型测定，其已经证明了B的直接可视化。在培养的螺旋体、血液样本和早期莱姆病患者的血液中，通过免疫荧光显微镜检测伯氏抗原阳性微生物。该项目的长期目标是使莱姆病的敏感性和特异性直接可视化实验室测试具有临床实用性。我们假设直接检测可视化分析将可靠和特异性地鉴定血液中的疏螺旋体螺旋体在低检测限。具体而言，我们将：（1）优化和验证测定的技术性能，并在每mL 2个生物体时达到80%的目标灵敏度;（2）评价背景信号，并在来自非流行性受试者的血液样品中进行加标研究，以定义具有90%特异性的阳性截止值，并建立85%的实验室内和实验室间重现性;（3）在试点研究中调查测定的临床性能特征，以包括来自莱姆病抗生素治疗之前、期间和之后的患者的样品。我们希望通过定义的评分标准和对照试剂实现优化和验证的检测，以直接检测B。在一项II期多实验室、盲法临床验证研究中，对血液中的伯氏螺旋体进行检测，以证明与目前的两级检测相比具有等效性或性能改善。该检测试剂盒的市场与莱姆病的高发病率相当，估计每年有100万至200万次检测。RareCyte将制定许可和合作策略，为FDA 510（k）批准提供试剂盒和仪器。

DESCRIPTION (provided by applicant): Lyme disease, a tick-borne infection caused by the spirochete Borrelia burgdorferi, has expanded to an estimated 300,000 US cases per year. Currently approved tests of immune response are sub-optimal and often give negative results early in the disease. The research outlined in this application is to develop a diagnostic blood test that directly visualizes B. burgdorferi. This test will allow earlier detection - and hence beter treatment - and could be used to monitor symptomatic patients for response to treatment. RareCyte Inc., an early-stage life science company dedicated to the characterization and retrieval of rare cells in blood, has a highly engineered enrichment technology based on a tube, float and collection mechanism that can stratify cells and pathogens in the blood by density. Using this technology, a prototype assay has been developed that has demonstrated direct visualization of B. burgdorferi antigen- positive organisms by immunofluorescence microscopy in cultured spirochetes, spiked- in blood samples, and in blood of patients with early Lyme disease. The long-term goal of this project is to make commercially available a sensitive and specific direct visualization laboratory test for Lyme disease with demonstrated clinical utility. We hypothesize that the direct detection visualization assay will reliably and specifically identif Borrelia spirochetes in the blood at a low limit of detection. Specifically, we will: (1) optimize nd validate technical performance of the assay and achieve target sensitivity of 80% at 2 organisms per mL; (2) evaluate background signal and perform a spike-in study in blood samples from non-endemic subjects to define a positive cut-off with 90% specificity, and establish intra- and inter-laboratory reproducibility of 85%; (3) investigate clinical performance characteristics of the assay in a pilot study to include samples from patients before, during and after Lyme disease antibiotic treatment. We expect to achieve an optimized and validated assay with defined scoring criteria and control reagents to directly detect B. burgdorferi in blood for ue in a Phase II multi-laboratory, blinded clinical validation study to demonstrate equivalence or improved performance over current two-tiered testing. The market for the assay parallels the high incidence of Lyme disease, and is estimated at between 1 and 2 million tests per year. RareCyte will develop licensing and partnering strategies to provide kits and instruments for FDA 510(k) approval.