Novel Point of Care assays for Urinary Diagnostics of Nephritis

用于肾炎尿液诊断的新型护理点检测

• 批准号: 9570651
• 负责人: CHANDRA MOHAN
• 金额: $ 34.2万
• 依托单位: UNIVERSITY OF HOUSTON
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-09-22 至 2021-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9570651
• 关键词: ALCAM gene; Address; Affect; African American; Anti-inflammatory; Antibodies; Biological Assay; Biological Markers; Biopsy; Biopsy Specimen; Blood; Calibration; Caucasians; Cellular Phone; Chinese People; Chronic; Clinic; Clinical; Clinical Research; Code; Computer software; Creatine; Creatinine; Detection; Development; Diagnosis; Diagnostic; Disease; Early Diagnosis; Enzyme-Linked Immunosorbent Assay; Equipment; Ethnic Origin; Ethnic group; Female; Female of child bearing age; Flare; Gold; Health; Health Services Accessibility; Hispanic Americans; Hispanics; Home environment; Immunoassay; Immunosuppressive Agents; Improve Access; Inflammation; Kidney; Laboratory Markers; Lateral; Latina; Longitudinal cohort; Lupus; Lupus Nephritis; Measurement; Measures; Methods; Modification; Monitor; Morbidity - disease rate; Nephritis; Organ; Outcome; Pathology; Patients; Performance; Personal Satisfaction; Pharmaceutical Preparations; Pregnancy Tests; Production; Properdin; Proteins; Public Health; Quality Control; Reading; Reagent; Reporter; Research; Sampling; Slide; System; Systemic Lupus Erythematosus; TFPI; Technology; Telephone; Testing; Time; Tissues; Urine; Validation; Woman; Work; aptamer; base; clinically actionable; cohort; diagnostic biomarker; improved; improved outcome; innovation; mortality; nano; nanoparticle; novel; novel marker; performance tests; point of care; predictive marker; protein biomarkers; prototype; screening; smartphone Application; systemic autoimmune disease; urinary; user-friendly

Project Summary/Abstract Systemic Lupus Erythematosus (“Lupus”) is a systemic autoimmune disease that leads to chronic inflammation in multiple organs including the kidneys. Patients commonly have exacerbations of the disease (kidney nephritis “flares”) interspersed by quiescent intervals. Early detection and the prompt treatment of flares can have a significant impact on the health and survival of patients, who are disproportionately female, Hispanic, and/or African-American. Biopsy sampling of kidney tissues is the gold standard for Lupus diagnosis, but is invasive and cannot be repeated frequently. In this context, blood or urine biomarkers that are predictive of kidney pathology could be very useful, especially if they supported point-of-care or ideally at-home testing. We have discovered and extensively validated lupus flare-correlated blood and urine diagnostic biomarkers in a protein screen of unprecedented scale, extending across 4 ethnic groups of patients (African American, Caucasian, Hispanic and Chinese). The urine markers perform better than conventional laboratory markers for Lupus and show potential to track with disease activity over time. While urine is an easy sample to give, urine biomarker concentrations must be corrected for their dilution by urine production. The molecule traditionally used for normalizing for urinary dilution, creatinine, is not very compatible with standard immunoassays, but we have identified a protein whose concentration in urine closely correlates with that of creatinine and that can be measured along with the flare markers. A point-of-care (Doctor's office), or better yet, a home self-test, for lupus flares could improve outcomes by expediting treatment. The natural format for such a test would be the lateral-flow assay (LFA), which is widely used as the home pregnancy test. Current LFAs, however, either require expensive equipment or lack the necessary sensitivity and quantitation. We propose to address this problem with smartphone-based LFAs based on our new phosphorescent (“glow-in-the-dark”) nanoparticle reporters. Motivating hypothesis: We hypothesize that the increased sensitivity and quantification ability of nanophosphor-LFAs will enable patients or doctors to simply measure our new kidney nephritis flare biomarkers in urine, and thus, address the unmet need for clinic/home monitoring of lupus nephritis flares. Successful completion of this work also will provide a generally-useful platform technology for quantitative smartphone LFA tests requiring no elaborate phone modifications (only a $10 slide-on attachment), and a new method of urine marker normalization well suited to general use in LFA and ELISA. Specific Aims: We propose: (Aim 1) To develop quantitative LFAs for urinary flare markers and creatinine-correlated normalizing protein; (Aim 2) To integrate multiple marker and normalizing protein tests into a user-friendly system with software error-catching, barcode reading, and quality controls; and finally (Aim 3) To evaluate the performance of the tests in lupus, using urine samples from broad cohorts of patients.

项目概要/摘要 系统性红斑狼疮（“狼疮”）是一种导致慢性炎症的系统性自身免疫性疾病 在包括肾脏在内的多个器官中，患者通常会出现病情恶化（肾脏）。 肾炎“发作”）中间有间歇期，可以早期发现并及时治疗发作。 对患者的健康和生存产生重大影响，患者中女性、西班牙裔、 和/或非洲裔美国人的肾组织活检取样是狼疮诊断的金标准，但并非如此。 在这种情况下，血液或尿液生物标志物具有侵入性且不能频繁重复。 肾脏病理学可能非常有用，特别是如果它们支持即时检测或理想的家庭检测。 已发现并广泛验证了狼疮发作相关的血液和尿液诊断生物标志物 规模空前的蛋白质筛查，涵盖 4 个种族的患者（非裔美国人、 白人、西班牙裔和中国人）尿液标记物的性能优于传统实验室标记物。 狼疮并显示出随着时间的推移跟踪疾病活动的潜力，虽然尿液是一种容易提供的样本，但尿液。 传统上，生物标志物浓度必须根据尿液产生的稀释程度进行校正。 用于标准化尿液稀释肌酐，与标准免疫测定不太兼容，但我们 已经确定了一种蛋白质，其在尿液中的浓度与肌酐的浓度密切相关，并且可以 与耀斑标记一起测量，或者更好的是，家庭自检。 狼疮发作可以通过加快治疗来改善结果。 然而，侧向层析检测 (LFA) 被广泛用作家庭妊娠测试。 需要昂贵的设备或缺乏必要的灵敏度和定量，我们建议解决这个问题。 基于我们新型磷光（“夜光”）纳米颗粒的智能手机 LFA 的问题 激励假设：我们已经建立了灵敏度和量化能力的提高。 纳米磷-LFA 将使患者或医生能够简单地测量我们新的肾炎发作 尿液中的生物标志物，从而解决了狼疮性肾炎发作的临床/家庭监测未得到满足的需求。 这项工作的成功完成也将为定量分析提供通用的平台技术。 智能手机 LFA 测试不需要复杂的手机修改（只需 10 美元的滑盖式附件），以及一个新的 尿液标记物标准化方法非常适合 LFA 和 ELISA 的一般用途 具体目标：我们。 建议：（目标 1）开发用于尿急发作标志物和肌酐相关正常化的定量 LFA 蛋白质；（目标 2）将标记物和标准化蛋白质测试多重集成到用户友好的系统中 软件错误捕获、条形码读取和质量控制；最后（目标 3）评估性能 使用来自广泛患者群体的尿液样本进行狼疮测试。