Detection of Biofilm Bacteria using GloBac

描述（由申请人提供）：使用目前的技术，感染关节成形术的微生物学诊断是不准确的。罗宾·帕特尔博士已经开发并验证了一项新技术，该技术可以对外植骨科植入物表面的生物膜中的微生物进行取样。该技术产生悬浮在溶液中的生物膜微生物；目前这些生物是用平板培养检测的。这种方法更敏感，但与假体周围组织培养一样特异性。然而，它不是快速的，因此不适合术中使用。术中假体关节感染诊断将有助于在手术室做出适当的手术决定，并且可能比目前使用的冷冻切片评估更快、更客观。采用新技术，生物膜微生物大量悬浮在溶液中（即100 ~ 200 cfu/ml）；因此，它们应该能够被直接发现。一种快速、自动化、灵敏、经济的悬浮生物膜细菌检测方法将使人工关节感染的快速诊断适用于术中应用。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对48%的敏感性，与假体周围组织的革兰氏染色（敏感性19%）相比有所提高，但仍不足以用于术中诊断。需要提高灵敏度。

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.