Multiplexed POC Diagnostic System for Bio-Threat Agents

描述（由申请人提供）：提出了一种多路护理点诊断系统（MPDS）的开发，用于快速诊断暴露于生物威胁剂的患者。该系统将执行多路测量，允许同时检测和识别液体（如血液）和拭子样本中的多种试剂。其他功能将包括对血液进行多重血清学检测，以检测对威胁因子的抗体反应。拟议的MPDS将包括两个要素：低成本、一次性墨盒和用于读取墨盒的小型便携式仪器。一次性，一次性使用的墨盒将包含进行分析所需的所有试剂，并将使用电化学发光检测。提出了两种药筒设计：一种用于测量液体样品中的分析物，包括全血和血浆，另一种用于提取和测量拭子上收集的分析物。墨盒将在小型便携式仪器上自动处理和分析，在不到15分钟的时间内提供结果。该计划的具体目标包括：(1)改进默沙杜的墨盒和墨盒阅读器的设计，并演示墨盒和阅读器的中试规模生产；(2)开发10种生物威胁因子的多重血清学和直接检测方法；(3)将化验板转移到先导墨盒和读取器；(4)在实际基质中优化检测试剂盒平台；(5)分析性能的广泛评价。在成功完成拟议的程序后，系统和分析将发展到可随后进入临床前和临床验证研究的水平。拟议的抗原和血清学检测小组包括以下10种威胁因子：炭疽芽孢杆菌（BA）、鼠疫耶尔森菌（YP）、土拉弗朗西斯菌（FT）、伯纳蒂克希菌（Q热）、布鲁氏菌病ssp、牛痘病毒、委内瑞拉马脑炎（VEE）、肉毒杆菌毒素A、蓖麻毒素和葡萄球菌肠毒素B （SEB）。除了这些目标外，为棉签样本开发的小组将包括对甲型流感、乙型流感和呼吸道合胞病毒的检测，以确定症状由常见呼吸道疾病引起的患者。相关性：迫切需要能够识别生物威胁物及其引起的疾病的诊断系统。拟议的诊断系统将通过提供具有中央实验室可靠性和性能的快速即时诊断来填补这一空白。该系统将能够快速识别暴露于生物威胁物或其毒素的个体，区分感染常见非威胁性上呼吸道疾病的个体，并允许回顾性血清学诊断以协助流行病学研究。该系统的设计目的是供未经专门培训的人员使用，并且不需要对样品进行预处理。

DESCRIPTION (provided by applicant): The development of a Multiplexed Point-of-Care Diagnostic System (MPDS) for rapid diagnosis of patients exposed to biological threat agents is proposed. The system will perform multiplexed measurements to allow for simultaneous detection and identification of multiple agents in both liquid (e.g., blood) and swab samples. Additional capabilities will include multiplexed serological testing of blood for antibody responses to threat agents. The proposed MPDS will consist of two elements: low cost, disposable cartridges and a small portable instrument for reading of cartridges. The disposable, single-use cartridges will contain all reagents necessary to conduct the assays and will use electrochemiluminescent detection. Two cartridge designs are proposed: one to measure analytes in liquid samples, including whole blood and plasma, and another to extract and measure analytes collected on swabs. The cartridges will be automatically processed and analyzed on a compact, portable instrument, providing results in less than 15 minutes. Specific goals of the program include: (1) refinement of MSD's cartridge and cartridge reader designs and demonstration of pilot scale production of both cartridges and reader; (2) development of multiplexed serology and direct detection assays for 10 biothreat agents; (3) transfer of the assay panels to the pilot cartridges and reader; (4) optimization of the assays on the cartridge platform in real matrices; and (5) extensive evaluation of assay performance. On successful completion of the proposed program, the system and assays will be developed to a level where they can subsequently be entered into pre-clinical and clinical validation studies. The proposed test panel for both antigen and serological tests include the following ten threat agents: Bacillus anthracis (BA), Yersinia pestis (YP), Francisella tularensis (FT), Coxiella burnetii (Q Fever), Brucellosis ssp, vaccinia virus, Venezuelan Equine Encephalitis (VEE), botulinum toxin A, ricin, and Staphylococcal enterotoxin B (SEB). In addition to these targets, the panel developed for swab samples will include assays for influenza A, influenza B, and RSV to identify patients whose symptoms are due to common respiratory ailments. Relevance: There is an urgent need for diagnostic systems capable of identifying biological threat agents and the diseases they cause. The proposed diagnostic system will fill that gap by providing rapid point-of-care diagnostics with the reliability and performance of the centralized laboratory. The system will be able to quickly identify individuals exposed to biological threat agents or their toxins, differentiate those individuals infected with common non-threat upper respiratory disease, and allow retrospective serological diagnosis to assist in epidemiological studies. The system is designed to be used by personnel without specialized training and will require no pre-processing of samples.