An antibiogram-based CentriCapillary system for neonatal sepsis PID and AST

 描述（由申请人提供）：本申报产品开发工作的主要目标是根据每个NICU在当地建立的流程图，使用全自动循证快速PID/AST系统，优化感染治疗并减少与抗生素使用不当相关的不良事件。在成功完成SBIR快速通道后，拟议的新生儿败血症CentriCapillary系统的最终目标是首先解决NICU环境中的分析需求，然后解决整个血液培养中的分析需求 市场（目前估计仅在美国就超过30亿美元）。新生儿不是小的成年人。设计使用10 mL成人血培养瓶和仅装载1 mL新生儿血样的诊断系统是不切实际的。毫无疑问，极低水平的菌血症存在错过靶病原体的风险（例如，低于1 CFU/mL），但常规从NICU的新生儿中获得超过1 mL血液用于PID和AST实验室检测是不实际的，甚至是不可能的。目前，主要的诊断公司还没有专门使用较小的血容量来解决新生儿败血症筛查的商业系统或产品开发工作。我们开发、展示并发表了一种创新的基于分子的基因型-表型-杂交方法，用于多重细菌PID和AST分析，在我们最近对73份原始临床尿液样本进行的临床可行性研究中，该方法具有100%的临床灵敏度、96%的临床特异性、98%的最小抑菌浓度（MIC）和97%的分类一致性。在这个fas track SBIR项目中，我们将利用我们在微流体，电化学生物传感器和临床微生物学方面的专业知识来开发集成的CentriCapillary系统。虽然我们的分子分析技术已经用引起UTI（这是最常见的细菌感染）的多药耐药病原体（MDR）进行了验证，但预计裂解-离心和动态杂交的结合将适用于快速新生儿败血症PID/AST和其他新兴感染性疾病。I期研究（第1年）具体目标1：将当前PID/AST平台技术从尿路感染（UTI）过渡到新生儿败血症-假设：裂解离心可通过使用血液的CentriCapillary柱来解决从原始尿液到全血样品的基质效应的变化以及新生儿败血症的低丰度病原体的问题。对新生儿的各种低血容量（10 μL - 2 mL）进行沉淀辅助快速血培养。具体目标二：证明基于电化学的分子分析CentriCapillary检测盒的可行性-假设：基于微流控芯片的血液沉淀可以捕获和回收YNHH NICU快速新生儿败血症PID和AST的免疫图上的所有常见病原体。II期研究（2年和3年）。具体目标3：开发一种基于双模式电化学的动态杂交分析算法，以扩展PID/AST面板-假设：动态杂交分析可用于扩展新生儿败血症常见和新兴病原体的种属特异性鉴定。具体目标4：原型，验证和制造CentriCapillary盒（COGS<$15）和系统（COGS<$25k）-假设：超离心（高达50，000 g，重力）可以结合到多路复用射流盒中，用于完全分离。 自动新生儿败血症PID/AST从全血样品中在6小时内。具体目标五：根据CLSI指南对快速新生儿PID/AST CentriCapillary系统进行临床验证-假设：分析验证研究中使用的加标ATCC菌株细菌的血液样本代表新生儿新鲜全血样本的关键基质特征。新生儿败血症的开发和验证将遵守ISO 13485标准，FDA/CDC/CLSI AST指南和联邦法规21 CFR 866.1645。本研究的最终目标是减少NICU环境中与抗生素使用不当相关的不良事件。

 DESCRIPTION (provided by applicant): The key objective of this proposed product development effort is to optimize the treatment of infections and reduce adverse events associated with inappropriate antibiotic use according to each NICU's locally established antibiogram with a fully automated, evidence-based rapid PID/AST system. The ultimate goal of the proposed neonatal sepsis CentriCapillary system after successful completion of this SBIR fast track is to initially address analysis needs in NICU settings then in the entire blood culture market (currently estimated to total more than $3 billion in the United States alone). Neonates are not small adults. It is not practical to design a diagnostics system using 10 mL blood culture bottles for adults and only load 1 mL of blood sample for neonates. Undoubtedly, there is a risk of missing the target pathogen for extremely low-level bacteremia (e.g., below 1 CFU/mL) using small blood sample volume, but it is not practical or even possible to obtain more than 1 mL of blood routinely from neonates in the NICU for PID and AST lab tests. Currently, there is no commercial system or product development effort from major diagnostics companies to address neonatal sepsis screening specifically using smaller blood volume. We developed, demonstrated and published an innovative molecular-based genotypic-phenotypic-hybrid approach for multiplexed bacterial PID and AST profiling with 100% clinical sensitivity, 96% clinical specificity, 98% minimum inhibitory concentration (MIC) and 97% categorical agreement in our most recent ongoing clinical feasibility study on 73 raw clinical urine samples. In this fas track SBIR project, we will leverage our expertise in microfluidics, electrochemical biosensors, and clinical microbiology to develop an integrated CentriCapillary system. While our molecular analysis technology has been validated with multidrugresistant pathogens (MDR) that cause UTI, which is the most common bacterial infection, it is anticipated that the incorporation of the lysis-centrifugation and dynamic hybridization will have applicability for rapid neonatal sepsis PID/AST and other emerging infectious diseases. PHASE I STUDY (YEAR 1) Specific Aim 1: Transition the current PID/AST platform technology from urinary tract infection (UTI) to neonatal sepsis- Hypothesis: Lysis centrifugation can address the change of matrix effect from raw urine to whole blood samples and the issue of low abundant pathogen for neonatal sepsis by using a CentriCapillary cartridge for blood- pelleting assisted rapid blood culture on various low blood volumes from neonates (10 μL - 2 mL). Specific Aim 2: Demonstrate the feasibility of the electrochemical-based molecular analysis CentriCapillary cartridge - Hypothesis: Microfluidic cartridge-based blood pelleting can capture and recover all common pathogens on the YNHH NICU's antibiogram for rapid neonatal sepsis PID and AST. PHASE II STUDY (YEARS 2 AND 3). Specific Aim 3: Develop a dual-mode electrochemical-based dynamic hybridization analysis algorithm to expand the PID/AST panel - Hypothesis: Dynamic hybridization analysis can be utilized to expand the species-specific identification of common and emerging pathogens for neonatal sepsis. Specific Aim 4: Prototype, validate and manufacture the CentriCapillary cartridge (COGS<$15) and system (COGS<$25k) - Hypothesis: Ultracentrifugation (up to 50,000 g, gravitational force) can be incorporated into a multiplexed fluidic cartridge for a fully automated neonatal sepsis PID/AST from whole blood samples in 6 hours. Specific Aim 5: Clinically validate the rapid neonatal PID/AST CentriCapillary system according to CLSI guidelines - Hypothesis: Blood samples spiked with ATCC strain bacteria used in the analytical validation studies represent critical matrix characteristics of fresh whole blood samples from neonates The development and validation of neonatal sepsis will adhere to the ISO 13485 standards, FDA/CDC/CLSI guidelines on AST and federal regulation 21 CFR 866.1645. The ultimate goal of this study is to reduce adverse events associated with inappropriate antibiotic use in the NICU settings.