Rapid sensitive low-cost test for resistant microbes causing hospital infections

描述（由申请人提供）：我们建议开发一种新的自动化平台，以实现快速，敏感，用户友好且具有成本效益的抗生素耐药细菌筛查，这些细菌经常导致严重的医疗相关感染。每年，约有10万人死于200万起与医疗保健相关的严重感染，估计造成的损失为350亿美元。大约2万人死于耐甲氧西林金黄色葡萄球菌（MRSA）。许多健康人都有MRSA定植。当这些MRSA携带者住院时，耐药微生物会对携带者和医院里的其他人造成严重感染。最近的几项研究表明，医院可以通过使用快速测试来筛选入院患者的MRSA定植，然后用严格的感染控制程序治疗定植的患者，从而显著减少医疗保健相关感染的数量。基于聚合酶链反应（PCR）的新型MRSA快速检测速度很快，但要么太贵，要么太复杂，无法在大多数医院实施。传统的微生物学方法用于筛查患者的MRSA并不昂贵，但太慢（1-2天），无法最有效地降低感染率。我们最近完成了一期项目，展示了一种新型、快速、简单、成本效益高的MRSA筛查方法的可行性，这将首次允许任何医院实施感染控制的快速检测。该平台技术使用低成本的非放大数字成像来快速计数单个荧光标记的金黄色葡萄球菌细胞。该测试通过检测金黄色葡萄球菌细胞来确定病人的样本是否含有MRSA，金黄色葡萄球菌细胞可以在含有抗生素的培养基中短暂孵育期间生长。快速（3.3小时）I期MRSA检测的分析灵敏度与PCR检测（10 CFU/样本）相当。当应用于临床鼻样本（n=163）时，该测试显示出与市场上的PCR测试相当的高临床敏感性（95%）和特异性（98%）。我们还设计、开发和集成了一个全功能的自动化台式分析仪原型，并开发和制造了一个用于MultiPath MRSA测试的低成本消耗品原型。在此，我们建议在第一阶段工作的基础上开发一个原型自动化平台和MRSA测试盒，可用于随后的第三阶段beta现场试验。具体而言，我们的目标是：(1)进一步缩短获得结果的时间并开发优化的稳定试剂；(2)开发低成本的MultiPath MRSA/SA检测试剂盒；(3)开发低成本自动化beta MultiPath分析仪；(4)整合和验证平台功能；(5)演示多路径MRSA/S的性能。金黄色葡萄球菌（MRSA/SA）试验。实现这些具体目标将使后续的III期beta现场测试成为可能，从而为未来的监管提交提供支持。MultiPath平台还将能够容纳广泛的检测菜单，用于引起医疗保健相关感染的病原体（例如，耐万古霉素肠球菌和艰难梭菌）、感染性病毒、食物病原体、毒素、分子疾病标记物和药物发现靶标。

DESCRIPTION (provided by applicant): We propose to develop a novel automated platform to enable rapid, sensitive, user-friendly, and cost-effective screening for the antibiotic resistant bacteria that frequently cause serious healthcare associated infections. Every year, about 100,000 deaths result from 2 million serious healthcare associated infections at an estimated cost of $35B. About 20,000 fatalities are caused by the bacterium methicillin-resistant Staphylococcus aureus (MRSA). Many healthy individuals are colonized with MRSA. When these MRSA carriers are hospitalized, the resistant microbe can cause serious infections for the carrier and others in the hospital. Several recent studies have shown that hospitals can significantly decrease the number of healthcare associated infections by using rapid tests to screen admitted patients for MRSA colonization and then treating the colonized patient with rigorous infection control procedures. New rapid MRSA tests based on PCR are fast, but are either too expensive or too complex to be implemented in most hospitals. Traditional microbiological methods for screening patients for MRSA are inexpensive but are too slow (1-2 days) to most effectively lower infection rates. We recently completed a Phase I project demonstrating the feasibility of a novel, rapid, simple, and cost-effective method for MRSA screening that should for the first time allow any hospital to implement rapid testing for infection control. The platform technology uses low-cost non-magnified digital imaging to rapidly count individual fluorescently labeled S. aureus cells. The test determines whether a patient's sample contains MRSA by testing for S. aureus cells that can grow in antibiotic-containing media during a brief incubation. The rapid (3.3 hr) Phase I MRSA test's analytical sensitivity is comparable to PCR tests (10 CFU/sample). When applied to clinical nasal samples (n=163), the test showed high clinical sensitivity (95%) and specificity (98%) comparable to the on-market PCR tests. We also designed, developed, and integrated a fully functional automated bench top analyzer prototype and developed and manufactured a low-cost consumable prototype for the MultiPath MRSA test. Here we propose to build on our Phase I work to develop a prototype automated platform and MRSA test cartridge that can be used in subsequent Phase III beta field trials. Specifically we aim to (1) further decrease time to results and develop optimized stable reagents; (2) develop a low-cost MultiPath MRSA/SA test cartridge; (3) develop a low-cost automated beta MultiPath analyzer; (4) integrate and verify the platform functionality; and (5) demonstrate the performance of the MultiPath MRSA/S. aureus (MRSA/SA) test. Achieving these Specific Aims will enable subsequent Phase III beta field testing that will in turn support a future regulatory submission. The MultiPath platform will also be able to accommodate a broad menu of tests for agents that cause healthcare associated infections (e.g., vancomycin resistant Enterococcus and C. difficile), infectious viruses, food pathogens, toxins, molecular disease markers, and drug discovery targets.