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，以实现对经常导致严重医疗保健相关感染的抗生素耐药性细菌的快速、敏感、用户友好和成本效益的筛选。每年约有10万人死于200万例严重的医疗保健相关感染，估计成本为350亿美元。约有20,000人死于耐甲氧西林金黄色葡萄球菌(MRSA)。许多健康的人都感染了MRSA。当这些MRSA携带者住院时，耐药微生物可能会对携带者和医院内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检测来筛选入院患者的MRSA定植，然后对定植的患者进行严格的感染控制程序，从而显著减少医疗保健相关感染的数量。基于聚合酶链式反应的新的快速MRSA检测速度很快，但要么太昂贵，要么太复杂，无法在大多数医院实施。传统的微生物学方法用于筛查MRSA患者成本低廉，但速度太慢(1-2天)，无法最有效地降低感染率。我们最近完成了一个第一阶段的项目，展示了一种新的、快速、简单和具有成本效益的MRSA筛查方法的可行性，这种方法应该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、非放大的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。这项检测通过检测金黄色葡萄球菌细胞在短暂的孵育过程中可以在含有抗生素的介质中生长来确定患者的样本是否含有MRSA。快速(3.3小时)第一阶段MRSA检测的分析灵敏度与聚合酶链式反应(10 cfu/样本)相当。当应用于临床鼻样(n=163)时，该检测显示出与市场上的聚合酶链式反应检测相当的高临床敏感性(95%)和特异性(98%)。我们还设计、开发和集成了全功能自动台式分析仪原型，并开发和制造了用于多路径MRSA测试的低成本消耗品原型。在这里，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。具体地说，我们的目标是(1)进一步缩短结果得出结果的时间，并开发优化的稳定试剂；(2)开发低成本多路径MRSA/SA测试盒；(3)开发低成本自动化贝塔测试仪；(4)集成和验证平台功能；以及(5)展示多路径MRSA/金黄色葡萄球菌(MRSA/SA)测试的性能。实现这些具体目标将使随后的第三阶段测试场测试成为可能，这反过来又将支持未来的监管提交。多路径平台还将能够对导致医疗保健相关感染(例如，万古霉素耐药肠球菌和艰难梭菌)、传染性病毒、食品病原体、毒素、分子疾病标志物和药物发现目标的制剂进行广泛的测试。

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.