Point-of-care concentration and detection of pathogen antigens in samples

描述(由申请人提供)：该项目将展示一种在设备最少的实验室或在护理地点(POC)诊断传染病的强大新策略。为生物防御和类似的低复杂性测试目的而开发的RAMP(R)抗原检测系统(加拿大卑诗省温哥华的Response Biomedical Corporation)将用于结核病(TB)的POC诊断。该匝道系统已经由独立的测试人员进行了广泛的评估，其高灵敏度和出色的稳健性已被证明。它的灵敏度将通过与一种新的电液动力微针系统的集成而得到提高，该系统旨在在RAMP分析之前选择性地将结核分枝杆菌(MTB)细胞集中在痰样本中。该微尖系统由NanoFacture，Inc.与华盛顿大学的纳米制造实验室合作开发和验证。集成的微尖-斜坡测试将检测结核分枝杆菌细胞，检测下限为每毫升痰100 cfu，与总测试时间20分钟的聚合酶链式反应相当。 该项目的具体目标是：1)开发痰中结核分枝杆菌细胞的Ramp测试；2)开发利用微尖系统浓缩人类痰中的MTB细胞的样品处理程序；3)在孟加拉国达卡的现场试验中评估微尖-Ramp系统的性能。与培养相比，20分钟微尖坡度试验的临床前目标表现具有90%的敏感性和特异性。此外，这一假设将得到检验，即微尖-坡度法明显比AFB涂片显微镜更敏感，AFB涂片显微镜是目前世界上使用最广泛的快速检测痰中结核杆菌细胞的方法。

DESCRIPTION (provided by applicant): This project will demonstrate a powerful new strategy for diagnosing infectious diseases in minimally equipped laboratories or at point of care (POC). Developed for biodefense and similar low-complexity testing purposes, the RAMP(R) antigen detection system (Response Biomedical Corporation, Vancouver, BC, Canada) will be adapted for POC diagnosis of tuberculosis (TB). The RAMP system has been extensively evaluated by independent testers, and its high sensitivity and outstanding robustness have been demonstrated. Its sensitivity will be enhanced by integration with a novel electrohydrodynamic microtip system designed to selectively concentrate Mycobacterium tuberculosis (MTB) cells in sputum samples prior to RAMP analysis. The microtip system was developed and validated by NanoFacture, Inc., in collaboration with the Nanomanufacturing Laboratory at the University of Washington. The integrated microtip-RAMP test will detect MTB cells with a lower limit of detection of <100 cfu per mL of sputum, comparable to PCR in a 20 min total test time. Specific aims of the project are: 1) To develop a RAMP test for MTB cells in sputum; 2) to develop sample processing procedures that utilize the microtip system to concentrate MTB cells from human sputum; and 3) to evaluate the performance of the microtip-RAMP system in a field trial in Dhaka, Bangladesh. Targeted pre-clinical performance for the 20-min microtip-RAMP test will have sensitivity and specificity of >90% relative to culture. In addition, the hypothesis will be tested that microtip-RAMP is significantly more sensitive than AFB smear microscopy, currently the world's most widely used rapid test to detect MTB cells in sputum.