Detection of Biofilm Bacteria using GloBac

描述（由申请人提供）：使用当前技术对关节置换术感染的微生物诊断不准确。Robin Patel博士开发并验证了一种新技术，该技术可对植入骨科植入物表面生物膜中的微生物进行采样。该技术产生悬浮在溶液中的生物膜微生物;目前这些生物体使用平板培养来检测。这种方法更敏感，但与假体周围组织培养一样特异。然而，它并不快速，因此不适合术中使用。术中人工关节感染诊断将使适当的手术决定，在手术室，并可能比目前使用的冷冻切片评价更快，更客观。使用新技术，生物膜微生物以高量悬浮在溶液中（即，>200 cfu/ml）;因此，它们应该适合于直接检测。用于检测悬浮生物膜细菌的快速、自动化、灵敏、具有成本效益的方法将使得能够进行适合于术中应用的快速假体关节感染诊断。Patel博士的研究小组已经证明，悬浮生物膜细菌的革兰氏染色的敏感性为48%，与假体周围组织革兰氏染色（敏感性19%）相比有所改善，但仍不足以用于术中诊断。需要提高灵敏度。 SUBC，Inc.正在开发一种新型的、灵敏的、全自动的、微处理器驱动的活细菌检测系统，称为“GloBac”。这种新技术被设计为基于细菌ATP的测量提供{90秒}灵敏和特异的细菌检测测试，并且预计是成本有效的。该I期STTR提案的目标是开发GloBac快速细菌检测系统，并确定该新型系统是否可以快速，灵敏和特异性地检测从人工关节感染患者中分离的嗜酸性和生物膜细菌。假设第一阶段可行，第二阶段将包括改进GloBac系统用于手术室和通过前瞻性测试接受膝关节或髋关节置换术翻修或切除的患者进行临床验证（即，不需要文化。该系统可能适用于其他临床标本的检测，包括但不限于从其他异物和滑液中脱落的生物膜细菌。公共卫生相关性：髋关节和膝关节置换术的频率越来越高;虽然大多数接受这种手术的患者都能缓解疼痛并恢复关节功能，但有些患者因感染或其他原因需要重复手术。失败的关节置换术的手术和医疗管理取决于是否存在感染;目前没有办法快速检测在手术室中接受失败的关节置换术的患者中引起感染的细菌。拟议研究的目标是开发一种用于检测人工关节上细菌的快速测试，可用于手术室诊断感染的人工关节。

DESCRIPTION (provided by applicant): The microbiologic diagnosis of infected joint anthroplasty is inaccurate using current techniques. Dr. Robin Patel has developed and validated a new technique that samples microorganisms in biofilms on the surface of explanted orthopedic implants. This technique yields biofilm microorganisms suspended in solution; currently these organisms are detected using plate culture. This approach is {more} sensitive, yet as specific as periprosthetic tissue culture. However, it is not rapid and therefore not amenable to intraoperative use. An intraoperative prosthetic joint infection diagnostic would enable appropriate surgical decisions to be made in the operating room, and could potentially be faster and more objective than the currently used frozen section evaluation. Using the new technique, biofilm microorganisms are suspended in solution in high quantity (i.e., >200 cfu/ml); they should therefore be amenable to direct detection. A rapid, automated, sensitive, cost-effective method for detection of suspended biofilm bacteria would enable a rapid prosthetic joint infection diagnostic amenable to intraoperative application. Dr. Patel s group has demonstrated that Gram stain of the suspended biofilm bacteria is 48% sensitive, which is improved compared to periprosthetic tissue Gram stain (sensitivity 19%), but still inadequate for an intraoperative diagnostic. Improved sensitivity is needed. SUBC, Inc. is developing a novel, sensitive, fully automated, microprocessor driven viable bacterial detection system called "GloBac". This novel technology is being designed to provide a {90 second} sensitive and specific bacterial detection test based on the measurement of bacterial ATP, and is anticipated to be cost-effective. The goal of this phase I STTR proposal is to develop the GloBac rapid bacterial detection system and to determine if this novel system can rapidly, sensitively and specifically detect planktonic and biofilm bacteria isolated from patients with prosthetic joint infection. Assuming feasibility in phase I, phase II will consist of refinement of the GloBac system for use in surgical suites and clinical validation by prospectively testing patients undergoing knee or hip arthroplasty revision or resection {i.e., without the need for culture}. This system is potentially applicable to testing of other clinical specimens including, but not limited to, dislodged biofilm bacteria from other foreign bodies, and synovial fluid. PUBLIC HEALTH RELEVANCE: Hip and knee replacements are being performed with increasing frequency; while most patients who undergo this type of surgery experience relief of pain and restoration of joint function, some require repeat surgery for infection or other reasons. Surgical and medical management of failed joint replacements depend on whether or not infection is present; there is currently no way to rapidly detect bacteria causing infection in the operating room in patients undergoing surgery for a failed joint replacement. The goal of the proposed studies is to develop a rapid test for detection of bacteria on artificial joints that can be used in the operating room to diagnose infected artificial joints.