VALIDATION OF A MULTIPLEXED ASSAY FOR BLADDER CANCER DIAGNOSIS

知识的差距：目前没有准确的，非侵入性的测试可用于血尿患者的规则 或有膀胱癌（BCa）病史，需要进行侵入性膀胱镜检查以评估膀胱是否存在 肿瘤背景：膀胱癌患者最常见的症状是血尿。在两个大 研究中，发现约11%的血尿患者携带BCa。发布的指南建议这些 除膀胱镜检查外，患者还需进行排尿细胞学检查（VUC）。膀胱镜检查是一种 与诸如短暂排尿副作用相关的侵入性、不舒服和昂贵的程序 症状、血尿、UTI和尿道狭窄，而VUC对膀胱癌的敏感性有限，为25-40%。 检测BCa（特异性>90%），特别是低级别和低阶段肿瘤。虽然一些商业 虽然可获得用于检测BCa的基于尿的测定，但许多测定存在降低的问题， 与VUC相比的特异性（例如，NMP-22和BTA）。此外，作为单一标记物，这些测定， 包括VUC的预测能力不足以应用于个体患者的管理， 重要的是，这些技术是复杂的，并且需要熟练的解释。我们目前的NIH/NCI R 01 应用是测试多重电化学发光（MEC）免疫测定检测我们的BCa的能力， 来自患有肉眼血尿的受试者和患有肉眼血尿的受试者的排泄尿液样品中的相关诊断特征 肿瘤监测中的BCa病史。自R 01应用程序开始以来，我们已经开发并 验证了具有改进操作特性的多重珠基（MBB）免疫测定法 如受试者操作特征下面积、敏感性和特异性（0.942：CI 0.8645 - 0.9627， 使用MBB平台分别为93%和95%，使用0.892：CI 0.850 - 0.934，分别为85%和81% MEC平台）。由于MBB测定法的实质性改进，我们寻求将 将MBB检测试剂盒评估纳入当前R 01授权，因此将MBB检测试剂盒与MEC进行比较 本补充申请中的分析。假设：MBB测定法的灵敏度和特异性高于 MEC检测我们在排尿样本中的BCa相关诊断特征。方法： 对于MEC测定，我们将在CLIA认证的实验室中进行MBB测定。我们将比较， 对比当前大型多中心中MBB和MEC检测的操作特征 前瞻性研究。在完成拟议的工作后，我们希望表明MBB测定法是非- 在检测我们的BCa特征方面不如MEC测定。因此，MBB检测将过渡到 实验室，并用于我们随后的研究，最终目标是加快翻译的步伐， 将我们的NCI支持的方法/测定/技术应用于临床，这是PAR-17-003的前提。

Gap in Knowledge: No accurate, non-invasive tests are currently available to rule in patients with hematuria or a history of bladder cancer (BCa) who need an invasive cystoscopy to evaluate the presence of a bladder tumor. Background: The most common presenting symptom in patients with BCa is hematuria. In two large studies, ~11% of patients with hematuria were noted to harbor BCa. Published guidelines recommend these patients to obtain voided urinary cytology (VUC), in addition to cystoscopic evaluation. Cystoscopy is an invasive, uncomfortable and expensive procedure associated with side effects such as transient voiding symptoms, hematuria, UTI, and stenosis of the urethra, whereas, VUC has limited sensitivity of 25-40% in detecting BCa (specificity is >90%), especially for low-grade and low-stage tumors. While some commercially available urine-based assays for the detection of BCa are available, many suffer from a reduction in assay specificity compared to VUC (e.g., NMP-22 and BTA). Furthermore, as single markers, these assays, including VUC have insufficient predictive power to be applied to the management of individual patients, and importantly, these techniques are complex, and require skillful interpretation. Our current NIH/NCI R01 application is testing a multiplex electrochemoluminescent (MEC) immunoassay’s ability to detect our BCa- associated diagnostic signature in voided urine samples from subjects with gross hematuria and subjects with a history of BCa on tumor surveillance. Since the inception of the R01 application, we have developed and validated a multiplex bead-based (MBB) immunoassay, which possesses improved operational characteristics such as area under receiver operating characteristic, sensitivity and specificity (0.942: CI 0.8645 – 0.9627, 93% and 95%, respectively using MBB platform vs. 0.892: CI 0.850 - 0.934, 85% and 81%, respectively using MEC platform). Because of the substantial improvement in the MBB assay, we seek to incorporate the evaluation of the MBB assay into the current R01 grant, and therefore compare the MBB assay to the MEC assay in this supplemental application. Hypothesis: The MBB assay is more sensitive and specific than the MEC assay in detecting our BCa-associated diagnostic signature in voided urine samples. Methodology: As with the MEC assay, we will perform the MBB assay in a CLIA-certified laboratory. We will then compare and contrast the operational characteristics of the MBB and MEC assays in the current, large multi-center prospective studies. Upon completion of the proposed work, we expect to show that the MBB assay is non- inferior to the MEC assay in detecting our BCa signature. Therefore, MBB assay will be transitioned into the laboratory and used for our subsequent studies with the ultimate goal of accelerating the pace of translation of our NCI-supported methods/assays/technologies to the clinic, which is the premise of PAR-17-003.