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

描述（由申请人提供）：该项目将展示在设备最低的实验室或护理点（POC）诊断传染病的强大新策略。RAMP(R)抗原检测系统（Response Biomedical Corporation, Vancouver， BC, Canada）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POC诊断结核病（TB）。RAMP系统已被独立测试人员广泛评估，其高灵敏度和出色的鲁棒性已被证明。通过与一种新型的电流体动力微尖系统集成，该系统可以在RAMP分析之前选择性地浓缩痰样本中的结核分枝杆菌（MTB）细胞，从而提高其灵敏度。该微针尖系统是由NanoFacture公司与华盛顿大学纳米制造实验室合作开发和验证的。集成的microtip-RAMP检测将检测MTB细胞，检测下限为<100 cfu / mL痰，与PCR相比，总检测时间为20 min。

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.