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型自动化平台，以实现快速、灵敏、用户友好和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保健相关感染。每年，约有10万人死于200万例严重的医疗相关感染，估计费用为350亿美元。大约有20,000人死于耐甲氧西林金黄色葡萄球菌（MRSA）。许多健康的个体被MRSA定殖。当这些MRSA携带者住院时，耐药微生物可导致携带者和医院其他人严重感染。最近的几项研究表明，医院可以通过使用快速测试来筛选住院患者的MRSA定植，然后用严格的感染控制程序治疗定植患者，从而显着减少医疗保健相关感染的数量。新的基于PCR的快速MRSA测试是快速的，但要么太昂贵，要么太复杂，无法在大多数医院实施。传统的微生物方法筛查MRSA患者是廉价的，但太慢（1-2天），最有效地降低感染率。我们最近完成了一个I期项目，证明了一种新的、快速的、简单的、具有成本效益的MRSA筛查方法的可行性，这将是第一次允许任何医院实施快速检测以控制感染。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S。金黄色葡萄球菌细胞该测试通过测试S来确定患者的样本是否含有MRSA。金黄色葡萄球菌细胞，其可以在短暂孵育期间在含有抗生素的培养基中生长。快速（3.3小时）阶段I MRSA测试的分析灵敏度与PCR测试相当（10 CFU/样品）。当应用于临床鼻样本（n=163）时，该测试显示出与市售PCR测试相当的高临床灵敏度（95%）和特异性（98%）。我们还设计、开发和集成了一个功能齐全的自动化台式分析仪原型，并开发和制造了一个用于多路径MRSA测试的低成本耗材原型。在这里，我们建议在我们的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β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检测盒;（3）开发低成本的自动化β MultiPath分析仪;（4）集成和验证平台功能;以及（5）证明MultiPath MRSA/S的性能。金黄色葡萄球菌（MRSA/SA）测试。实现这些特定目标将使后续III期β现场测试成为可能，进而支持未来的监管申报。MultiPath平台还将能够适应广泛的测试菜单，用于导致医疗保健相关感染的病原体（例如，万古霉素耐药肠球菌和C.艰难梭菌（difficile）、感染性病毒、食物病原体、毒素、分子疾病标志物和药物发现靶标。

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.