Culture-free identification of pathogens common in endocarditis directly from patient specimens

项目总结 感染性心内膜炎(IE)是一种危及生命的疾病，每年影响大约45,000名美国人。IE是 以心内膜表面的微生物植被为特征的，会导致严重的心脏损害和 持续性血流感染(BSI)导致败血症。如果不及时治疗，IE是致命的。在以下标准中 建立IE，最有效地治疗患者，血培养是鉴定病原体必不可少的。 然而，文化表现出两个主要的弱点，推迟了适当的抗菌剂的使用： (1)周转时间长达数天甚至数周，以及(2)由于以下原因之一而导致的假阴性结果的高流行率 用抗生素进行前处理或存在难以培养的病原体。因为时间对于最优是至关重要的 结果并发症最少，通常在诊断前用广谱的鸡尾酒开始治疗。 光谱(即非靶向)抗菌药，导致大多数患者治疗不当， 没有不必要的疾病治疗。因此，至关重要的是推进创新的诊断方法， 它们不依赖于培养，以便迅速过渡到个性化的抗微生物干预措施。 为了解决这一未得到满足的需求，HelixBind将开发第一个能够识别 在2.5小时内诱发IE的病原体。HelixBind的方法利用了来自 样本到答案。此外，由于检测不需要培养，我们的方法特别适合于 对挑剔的病原体的检测，这是当今的诊断标准经常无法检测到的。我们的建议 感染性心内膜炎病原体鉴定，被称为IE/PID，有望具有深远的临床意义 通过使医生能够在几小时内应用个性化和循证干预， 而不是几天；最终改善结果和抗菌药物管理。以前，HelixBind 演示了核心病原体识别(C/PID)平台，目标是处于危险中的普通患者群体 用于血液感染(BSI)引起的败血症。C/PID检测21种最常见的引起BSI的病原体 在单次检测中直接取血，从样品到答案只需不到2.5小时，并显示单次 整个面板上的cfu/ml LOD。C/Pid在一项使用临床标本的盲法先导研究中得到验证 并显示&gt；95%的敏感性和&gt；%的特异性。IE/PID将使用与C/PID类似的流程，并将 检测与IE相关的最普遍的病原体，包括挑剔和不可培养的病原体。 我们的具体目标和可量化的交付成果旨在证明在时间和 第一阶段的资本限制为了成功，我们组建了一支化验开发和 人工核酸，由领先的临床微生物学家提供指导。已经实现了我们的特定目标 目标，我们将在第二阶段开发可部署的仪器/消耗品，扩大测试面板以包括 通常被认为是不可培养的病原体，如伯氏柯克斯体和巴尔通体，并验证和 在一项盲法研究中，用临床标本验证自动诊断平台。

PROJECT SUMMARY Infective Endocarditis (IE) is a life-threatening disease that affects roughly 45,000 Americans annually. IE is characterized by microbial vegetations on the endocardial surface which induce serious heart damage and persistent bloodstream infections (BSIs) leading to sepsis. Left untreated IE is fatal. Among the criteria to establish IE and treat patients most effectively, blood cultures are indispensable to identify the etiologic agent. Cultures, however, display two major weaknesses which delay the administration of the proper antimicrobials: (1) High turnaround time of days or even weeks and (2) high prevalence of false-negative results due to either pre-treatment with antibiotics or the presence of difficult to culture pathogens. As time is of essence for optimal outcomes with minimal complications, treatment is often initiated prior to diagnosis with a cocktail of broad spectrum (i.e. not targeted) antimicrobials, leaving the majority of patients treated inappropriately and those without the disease treated unnecessarily. It is therefore critical to advance innovative diagnostic approaches, which do not rely on culturing, in order to rapidly transition to personalized antimicrobial interventions. To address this unmet need, HelixBind will develop the first culture-free diagnostic assay capable of identifying pathogens which induce IE in under 2.5 hours. HelixBind’s approach leverages multiple novel innovations from sample-to-answer. Further, as detection does not require cultures, our approach is particularly suited to the detection of fastidious pathogens, which today’s diagnostic standard frequently fail to detect. Our proposed Infective Endocarditis Pathogen Identification assay, termed IE/PID, is expected to have a profound clinical impact by enabling the physician to apply a personalized and evidence-driven intervention within a few hours, rather than days; ultimately improving both outcomes and antimicrobial stewardship. Previously, HelixBind demonstrated a Core Pathogen Identification (C/PID) platform, aimed at the ‘general’ patient population at-risk for sepsis caused by bloodstream infections (BSIs). C/PID detects 21 of the most prevalent BSI-causing pathogens in a single test directly from blood, requires less than 2.5 hours from sample-to-answer, and displayed single CFU/ml LODs across the entire panel. C/PID was validated in a blinded pilot study utilizing clinical specimens and demonstrated >95% sensitivity and >89% specificity. IE/PID will utilize similar processes as C/PID and will detect the most prevalent pathogens associated with IE including fastidious and uncultureable pathogens. Our Specific Aims with quantifiable deliverables are designed to demonstrate feasibility within the time and capital constraints of a Phase I. To succeed, we have assembled a team of experts in assay development and artificial nucleic acids, supported by leading clinical microbiologists for guidance. Having achieved our Specific Aims, we will, in Phase II, develop deployable instrumentation/consumables, expand the test panel to include pathogens generally deemed as ‘unculturable’, such as Coxiella burnetii and Bartonella spp., and verify and validate an automated diagnostic platform with clinical specimens in a blinded study.