Rapid Low Cost Phenotypic Antibiotic Susceptibility Testing.

快速低成本表型抗生素敏感性测试。

• 批准号: 9347745
• 负责人: PAUL RHODES
• 金额: $ 99.81万
• 依托单位: SPECIFIC DIAGNOSTICS, INC.
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-02-07 至 2020-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9347745
• 关键词: Affect; Agitation; Algorithms; Antibiotic Resistance; Antibiotic susceptibility; Antibiotics; Blood; Blood specimen; CLIA certified; Centers for Disease Control and Prevention (U.S.); Cessation of life; Characteristics; Chemicals; Clinical; Clinical Protocols; Clinical Research; Computer software; Consultations; Cyclosporine; Data; Dependence; Dependency; Detection; Development; Engineering; Environment; Evaluation; Future; Goals; Gold; Growth; Guidelines; Hand; Healthcare; Healthcare Systems; Hour; Incubators; International; Laboratories; Length of Stay; Liquid substance; Measures; Methods; Minimum Inhibitory Concentration measurement; Morbidity - disease rate; National Institute of Allergy and Infectious Disease; Organism; Pathogenicity; Patients; Pattern; Performance; Phase; Phenotype; Pilot Projects; Preclinical Testing; Predictive Analytics; Predisposition; Preparation; Protocols documentation; Research; Research Design; Resistance; Sampling; Shock; Speed; System; Systems Development; Technology; Temperature; Testing; Time; Walking; cost; cost effective; disorder prevention; engineering design; image processing; instrument; mortality; pathogen; pathogenic bacteria; prototype; research clinical testing; response; seal; sensor; skills; software systems; targeted treatment

Project Summary Antibiotic resistance has recently emerged as a global threat for healthcare systems. An increasing number of pathogenic bacteria are acquiring antibiotic resistance, and new forms of resistance are continuously emerging with alarming speed across international boundaries.1 In the U. S. alone, the Center for Disease Control and Prevention (CDC) has estimated 2 million patients per year are directly affected by antibiotic-resistant pathogens, leading to more than 23,000 deaths.2 Providing rapid, definitive and cost effective antibiotic susceptibility testing (AST) will be of increasingly vital importance in controlling this burgeoning problem. While legacy methods generally require overnight culturing, the time-urgency of determining effective antibiotics has prompted a push for rapid AST that can perform in a few hours. Unfortunately, some of the newest rapid AST instruments are either not suited to multiplexing, or are an order of magnitude more costly than legacy methods, discouraging their widespread use. A rapid multiplexed AST platform that provides minimum inhibitory concentrations (MICs) within hours at comparable cost to current methods will enable rapid determination of effective targeted therapy, resulting in short hospital stays and fewer additional laboratory tests, and decreasing morbidity and mortality along with associated healthcare expenses. Specific Technologies has introduced and with NIAID support commercially developed a culture system that combines detection with organism ID.3, 4 Using a printed array of chemically responsive colorimetric indicators introduced into culture headspace, the species-specific pattern of volatiles emitted during growth produce a species-specific pattern, which we have shown to reduce detection time and enable Gram status and species ID to be determined hands-free during culture. Here, we present data showing that the use of this system can be extended to very rapidly, order 1 hour, ascertain phenotypic antibiotic susceptibility. The preliminary data demonstrate that introduction of antibiotic to positive blood culture prompts a susceptibility-dependent and antibiotic concentration-dependent shift in volatile sensor response within <60 minutes. Recognizing that a commercial AST instrument must be multi-well, we have performed pilot study in which the positive sample is removed from a blood culture bottle and divided into a multi-chamber setting, in which even at pilot study stage we demonstrate a < 2 ½ hrs phenotypic AST, measuring values matching the results of the CLIA-compliant Sensititre gold standard. Moreover, the disposable is inherently low cost, as it only depends upon an inexpensive printed sheet of CSA’s. Finally, not only is this method intrinsically low-cost but low-skill compatible, as there is no sample preparation from positive blood culture. It is thus very well-suited for both 24-hour/day developed world labs, where it will speed time-to-answer, as well as the low-skill environments of the developing world. In this direct-to-Phase II application, we propose to use our CSA technology to systematically quantify, and validate the parameter-dependence of this system and then engineer and produce a benchtop instrument suitable for preclinical and clinical testing, which will provide a low cost, highly automated multiplexed phenotypic susceptibility determination directly from a positive blood sample in less than 90 minutes. We propose to develop and commercialize an instrument that integrates our rapid AST capability with our Spec80 blood culture and species ID system that is being commercialized now, offering a uniquely streamlined and low cost system with by far the fastest yet proposed time from patient sample to detection, ID and phenotypic multiplexed quantitative AST.

项目摘要 抗生素耐药性最近已成为医疗保健系统的全球威胁。越来越 许多病原菌正在获得抗生素耐药性，新的耐药性形式正在出现。 在美国，这种现象正以惊人的速度跨越国界。S.单 疾病控制和预防中心（CDC）估计，每年有200万患者直接 受抗药性病原体影响，导致23，000多人死亡。 确定的和成本有效的抗生素敏感性测试（AST）将越来越重要 来控制这个迅速发展的问题。虽然传统方法通常需要过夜培养，但 确定有效抗生素的时间紧迫性促使人们推动快速AST， 几个小时不幸的是，一些最新的快速AST仪器要么不适合 多路复用，或者比传统方法成本高一个数量级，阻碍了它们的应用。 广泛使用。提供最低抑制浓度的快速多重AST平台 （中等收入国家），将能够快速确定有效的 靶向治疗，缩短住院时间，减少额外的实验室检查， 发病率和死亡率沿着以及相关的医疗费用。 Specific Technologies在NIAID的支持下推出了一种商业化的培养系统， 将检测与微生物ID相结合。3，4使用印刷的化学响应比色法阵列， 指示剂引入培养物顶空，在培养过程中释放的挥发物的物种特异性模式 生长产生一种物种特异性模式，我们已经证明，这可以减少检测时间， 培养过程中，革兰氏状态和种属ID无需手动确定。在这里，我们提供的数据显示， 该系统的使用可以扩展到非常迅速，1小时，确定表型抗生素 易感性初步数据表明，将抗生素引入阳性血培养 促使挥发性传感器发生灵敏度依赖性和抗生素浓度依赖性变化 60分钟内回复。认识到商业AST仪器必须是多孔的，我们 进行了初步研究，其中从血培养瓶中取出阳性样本并将其分开 在多室环境中，即使在初步研究阶段，我们也证明了< 2 ½小时的表型 AST，测量值与符合CLIA标准的Sensititre金标准的结果相匹配。此外，委员会认为， 一次性用品本身成本低，因为它仅依赖于CSA的廉价印刷片。 最后，这种方法不仅本身成本低，而且与低技能兼容，因为没有样本 从阳性血培养中制备。因此，它非常适合24小时/天的发达国家 实验室，在那里它将加快回答时间，以及发展中国家的低技能环境。 在这个直接进入第二阶段的应用中，我们建议使用我们的CSA技术来系统地量化， 并验证了该系统的参数依赖性，然后设计并生产了一个实验台 仪器适用于临床前和临床测试，这将提供低成本，高度自动化 直接从阳性血液样本中进行多重表型易感性测定， 分钟我们建议开发和商业化的仪器，集成我们的快速AST 我们的Spec 80血液培养和物种识别系统目前正在商业化， 一个独特的流线型和低成本的系统，迄今为止最快的，但从病人样本的建议时间 用于检测、ID和表型多重定量AST。