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

描述（由申请人提供）：该项目将展示一种强大的新策略，用于在设备最少的实验室或护理点（POC）诊断传染病。RAMP（R）抗原检测系统（Response Biomedical Corporation，温哥华，BC，加拿大）是为生物防御和类似的低复杂性检测目的而开发的，将适用于结核病（TB）的POC诊断。RAMP系统已经过独立测试人员的广泛评估，其高灵敏度和出色的鲁棒性得到了证明。它的灵敏度将通过与一种新的电流体动力学微尖系统集成来增强，该系统设计用于在RAMP分析之前选择性地浓缩痰液样品中的结核分枝杆菌（MTB）细胞。微尖端系统由NanoFacture，Inc.开发和验证，与华盛顿大学的纳米制造实验室合作。集成的microtip-RAMP测试将检测MTB细胞，其检测下限为<100 cfu/mL痰液，与PCR在20分钟的总测试时间内相当。 该项目的具体目标是：1）开发痰液中MTB细胞的RAMP检测; 2）开发利用微尖系统从人痰液中浓缩MTB细胞的样本处理程序; 3）在孟加拉国达卡的田间试验中评价微尖-RAMP系统的性能。相对于培养物，20分钟微尖-RAMP测试的目标临床前性能将具有>90%的灵敏度和特异性。此外，还将检验微尖-RAMP比AFB涂片镜检法（目前世界上最广泛使用的检测痰液中MTB细胞的快速检测法）显著更敏感的假设。

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.