Near patient molecular diagnostics test for infections

摘要 目前，还没有FDA批准的基于分子的测试来鉴定尿路感染(UTI)病原体和 药敏试验将取代尿检和尿液培养的“黄金标准”。所有病原体 鉴定试验仍然依赖于尿液培养的临床分离株，与科赫提出的假设基本相同 在19世纪作为识别病原体的一般指南。然而，随着时间的推移，发生了变化的是戏剧性和 在这些病原体中逐渐出现抗生素耐药性。来自美国疾病控制和预防中心的数据 预防(CDC)国家医疗安全网(NHSN)表明，从 医院获得性感染(HAI)现在至少对两种主要类别的抗生素具有耐药性。最近的事态发展 多药耐药(MDR)病原体的分子诊断检测可以提供敏感、特异和实时的 支持主动监测驱动的感染控制干预措施的解决方案，但这些基于PCR的检测只能 在简单的标本基质上进行，如拭子或阳性血培养基(血与肉汤的比例为1：10)。不是 FDA批准了基于分子的方法，可以直接对尿样或全血样本进行检测。 这项商业化准备试点(CRP)赠款的目标是加快 我们的NIAID SBIR R44AI088756项目的计划成果，该项目名为“用于快速的集成诊断系统” 药敏试验(AST)“。我们建议在监管战略制定和管理方面提供技术援助 具有成本效益的制造，以及关于独立复制关键字的后期研究和开发活动 符合食品和药物管理局要求和国际标准化组织13485标准的研究。我们NIAID先进技术的目标 SBIR项目是为了开发和验证RAST(快速抗菌药敏感性检测)，一个集成和紧凑的 诊断系统，使临床医生能够通过基于证据的抗生素选择来指导护理点(POC) 治疗急性细菌感染。我们的第一个紧凑型自动化系统能够快速识别病原体(ID) 和AST直接从患者样本中获得循证信息，开始针对患者的抗菌治疗。 预计在2016年2月即将到来的国际标准化组织13485监督认证审核后，该公司将获得CE标志。然而， FDA的批准受到以下因素的阻碍：(1)FDA没有批准尿路感染的基于分子的预测测试 (2)传统试纸尿检的低成本，以及(3)通过多项检测缺乏新鲜尿样. 现场临床可行性研究。 我们将通过以下目标克服这些障碍： 技术援助 目标1：通过向外部设备提出从头开始请求，获得基于风险的II类设备分类 NSF国际(一家国际认证组织)在18个月内提供监管援助 目标2：降低墨盒销售成本3美元，外部制造系统销售成本5,000美元 发展援助 后期研发活动 目标3：进行多中心临床表现研究并演示&gt；95%的临床表现 敏感度/特异度和&gt；95%敏感度分类一致 在这项研究中，我们重点验证根据联邦法规21CFR 866.2660的尿路感染快速诊断设备 (微生物鉴定装置)、21CFR 866.1640(药敏试验粉)、 和21CFR 866.1645(全自动、短期孵化周期药敏检测系统)。美国食品和药物管理局 这类系统的产品代码是“JSS”和“LON”。监管战略将在目标1(监管)中最终确定为 在目标3(临床研究)中准备临床方案和多中心临床表现研究，将利用 目标2(制造)中优化的具有成本效益的系统和消耗品。

ABSTRACT Currently, there are no FDA cleared molecular-based tests for urinary tract infection (UTI) pathogen identification and antimicrobial susceptibility testing to replace the 'gold standard' of dipstick urinalysis and urine culturing. All pathogen identification tests still rely on clinical isolates from urine cultures, largely unchanged from Koch's postulates developed in the 19th century as general guidelines to identify pathogens. What has changed over time, however, is the dramatic and progressive emergence of antibiotic resistance among these pathogens. Data from the Centers for Disease Control and Prevention (CDC) National Healthcare Safety Network (NHSN) indicate that 70% of the pathogens isolated from hospital-acquired infections (HAIs) now are resistant to at least 2 major classes of antibiotics. Recent developments in molecular diagnostic testing for multidrug resistant (MDR) pathogens can provide a sensitive, specific, and real-time solution to support active surveillance-driven infection control interventions, but these PCR-based tests can only be performed on simple specimen matrices such as swabs or positive blood culture media (1:10 blood-to-broth ratio). No molecular-based method is cleared by the FDA to test directly on urine or whole blood samples. The goal of this Commercialization Readiness Pilot (CRP) Grant is to accelerate the commercialization effort of the program outcome from our NIAID SBIR R44AI088756 project titled “An Integrated Diagnostic System for Rapid Antimicrobial Susceptibility Testing (AST)”. We propose technical assistance on regulatory strategy development and cost-effective manufacturing, as well as later stage research and development activities on independent replication of key studies in compliance with FDA requirements and ISO 13485 standards. The goal of our NIAID Advanced Technology SBIR project is to develop and validate RAST (rapid antimicrobial susceptibility testing), an integrated and compact diagnostic system that enables clinicians direct point-of-care (POC) with an evidence-based selection of antibiotics for treatment of acute bacterial infections. Our first compact automated system is capable of rapid pathogen identification (ID) and AST directly from patient's samples with evidenced-based information to start patient-specific antimicrobial treatment. It is expected to obtain a CE Mark after the coming ISO 13485 surveillance certification audit in February 2016. However, the FDA clearance is hindered by: (1) no molecular-based predicate test cleared by the FDA for urinary tract infections (UTI), (2) the low cost of conventional dipstick urinalysis tests, and (3) the lack of fresh urine specimens through a multi- site clinical feasibility study. We will overcome these hurdles through the following aims: TECHNICAL ASSISTANCE Aim 1: Obtain a risk-based classification of the Class II device through a de novo request with external regulatory assistance from NSF International (an international certification organization) in 18 months Aim 2: Reduce the cost of goods sold of <$3 for cartridge and <$5k for system with external manufacturing development assistance LATE STAGE RESEARCH AND DEVELOPMENT ACTIVITIES Aim 3: Conduct a multi-center clinical performance study and demonstrate >95% clinical sensitivity/specificity and >95% susceptibility categorical agreement In this study, we focus on validating the rapid UTI diagnostic device according to federal regulations 21 CFR 866.2660 (microorganism differentiation and identification device), 21 CFR 866.1640 (antimicrobial susceptibility test powder), and 21 CFR 866.1645 (fully automated, short-term incubation cycle antimicrobial susceptibility testing system). The FDA product codes for such systems are “JSS” and “LON.” The regulatory strategy will be finalized in Aim 1 (Regulatory) for preparation of the clinical protocol and multicenter clinical performance study in Aim 3 (Clinical study) that will utilize the cost-effective system and consumables optimized in Aim 2 (Manufacturing).