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

 描述(由申请人提供)：这项拟议的产品开发工作的关键目标是根据每个NICU在当地建立的具有全自动、基于证据的快速PID/AST系统的抗生素图谱，优化感染的治疗，并减少与不适当使用抗生素相关的不良事件。在成功完成SBIR快速通道后，拟议的新生儿败血症中心毛细血管系统的最终目标是首先满足NICU环境中的分析需求，然后是整个血液培养 市场(目前估计仅在美国就超过30亿美元)。新生儿不是小个子成年人。设计一个成人用10毫升血培养瓶，新生儿只装1毫升血样的诊断系统是不现实的。毫无疑问，使用少量血液样本有可能遗漏极低水平菌血症(例如低于1 cfu/ml)的目标病原体，但从NICU中的新生儿常规获取超过1毫升的血液用于PID和AST实验室测试是不切实际的，甚至是不可能的。目前，主要的诊断公司还没有商业系统或产品开发工作来解决专门使用较小血量的新生儿败血症筛查问题。我们开发、论证并发表了一种创新的基于分子的基因-表型-混合方法，用于多重细菌PID和AST图谱分析，具有100%的临床敏感性、96%的临床特异性、98%的最低抑菌浓度(MIC)和97%的分类符合率。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在引起尿路感染的多重耐药病原体(MDR)中得到验证，这是最常见的细菌感染，但预计裂解-离心法和动态杂交的结合将适用于快速新生儿败血症、PID/AST和其他新出现的传染病。第一阶段研究(1年)具体目标1：将当前的PID/AST平台技术从尿路感染过渡到新生儿败血症-假设：裂解离心法可以解决从原始尿样到全血样本的基质效应变化以及新生儿败血症病原菌低丰度的问题，使用Centrian毛细管柱对不同低血量(10μL-2毫升)的新生儿进行快速血液培养。具体目标2：论证基于电化学的分子分析中心毛细管柱的可行性假设：基于微流控毛细管柱的血液造粒可以捕获和恢复YNHH NICU对新生儿快速败血症、PID和AST的抗生素图谱上的所有常见病原体。第二阶段研究(第二年和第三年)。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小组-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物种特异性鉴定。具体目标4：原型、验证和制造中央毛细管柱(COGS&lt；$15)和系统(COGS&lt；$25k)-假设：超速离心法(高达50,000 g，重力)可以集成到多路复用的流体柱中 在6小时内从全血样本中自动获取新生儿败血症的PID/AST。具体目标5：根据CLSI指南在临床上验证快速新生儿PID/AST中心毛细管系统-假设：分析验证研究中使用的添加ATCC菌株细菌的血液样本代表了新生儿新鲜全血样本的关键基质特征新生儿败血症的发生和验证将遵循ISO 13485标准、FDA/CDC/CLSI关于AST的指南和联邦法规21 CFR 866.1645。这项研究的最终目标是减少与NICU环境中不适当使用抗生素相关的不良事件。

 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.