Real-Time Reagent-Free Noninvasive Diagnosis of Tuberculosis

结核病的实时无试剂无创诊断

• 批准号: 8495922
• 负责人: David Philip Fergenson
• 金额: $ 29.6万
• 依托单位: LIVERMORE INSTRUMENTS, INC.
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-07-01 至 2015-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8495922
• 关键词: Accident and Emergency department; Acid Fast Bacillae Staining Method; Address; Aerosols; Algorithms; Anus; Area; Automated Pattern Recognition; Bacillus (bacterium); Bacillus anthracis; Bacterial Pneumonia; Beds; Border Crossings; Breath Tests; Bronchiectasis; Cells; Centers for Disease Control and Prevention (U.S.); Characteristics; Cicatrix; Clinic; Clinical; Clinical Engineering; Collection; Complex; Contracts; Cost Control; Coughing; Custom; Data; Data Analyses; Department of Energy; Detection; Development; Devices; Diagnosis; Diagnostic; Disease; Electricity; Engineering; Ensure; Environment; Evaluation; Exhalation; Funding; Future; Generations; Genus Mycobacterium; Gold; Health; Hospitals; Hour; Human; Incidence; Individual; Infection; Infection Control; Inpatients; Institution; Ions; Island; Lasers; Legal patent; Libraries; Licensing; Logistics; Lung diseases; Malignant neoplasm of lung; Masks; Mass Screening; Mass Spectrum Analysis; Measures; Medical Technology; Methods; Mycobacterium tuberculosis; Patients; Pattern Recognition; Performance; Phase; Physicians; Physiologic pulse; Polymerase Chain Reaction; Population; Prevention Guidelines; Process; Public Health; Pulmonary Tuberculosis; Reagent; Reproducibility; Research; Risk; Route; Safety; Sampling; San Francisco; Savings; Sea; Services; Shipping; Ships; Source; Sputum; Symptoms; System; Systems Analysis; Technology; Test Result; Testing; Time; Training; Trees; Triage; Tuberculosis; Uncertainty; United States; Update; Validation; base; clinically relevant; cost; cost effective; disorder prevention; experience; genome sequencing; instrument; instrumentation; interest; killings; mass spectrometer; metabolomics; microorganism; mycobacterial; noninvasive diagnosis; particle; pathogen; pressure; prevent; protocol development; respiratory; screening; tool; transmission process; user-friendly

DESCRIPTION (provided by applicant): Despite the human cost of tuberculosis (TB), the disease remains difficult to diagnose and thus to control. One-third of the world's population is infected with M. tuberculosis. Every year, more than 9 million people develop active TB and 1.7 million people die from the disease [WHO 2010]. Current widely used diagnostics perform poorly in identifying active TB in patients with roughly 50% of cases having sputa that are smear negative for acid fast bacilli (AFB) [Steingart 2006]. Undiagnosed cases unwittingly continue to transmit disease, threatening public health. In the US, the uncertainty in diagnosis, and the absence of sensitive, specific, and rapid tests, results in significant increases in hospitalizatio costs because Centers for Disease Control and Prevention (CDC) guidelines recommend that suspected cases of TB be isolated in single bed, negative-pressure rooms until three consecutive daily sputum samples have been processed by direct smear and deemed negative for AFB [CDC 2005]. Due to logistics issues, these patients remain in isolation an average of five days before being cleared. Because the vast majority of these patients do not have the disease, TB control cost at the inpatient level is dominated by these inpatient isolation days for TB suspects [Scott 1994] and is often ultimately borne by the hospital itself. Compounding this infection control approach is the fact that negative sputum smears for AFB do not definitively exclude risk of TB transmission [Behr 1999]. Societal costs increase dramatically when undiagnosed cases continue to transmit disease [Miller 2010]. The limitations of the current gold standards for TB diagnosis have stimulated renewed interest in rapid, accurate, cost-effective, non-invasive, and user-friendly methods of identifying M. tuberculosis from TB suspects. Patient cough aerosol analysis for M. tuberculosis has the potential to address all these requirements. Livermore Instruments' unique Single Particle Aerosol Mass Spectrometry (SPAMS) technology may be the answer to TB diagnosis in clinical real-world settings. SPAMS has been previously deployed by the Department of Energy for the detection of B. anthracis. In this application, we describe an aerosol analysis system based on SPAMS as a potential tool for detection of M. tuberculosis in coughed and exhaled breath. The existing versions of our system are simple to operate, sturdy, contain few moving parts, and, importantly, are reagent-free so that the marginal cost per test is extremely low. SPAMS functions by drawing in individual aerosol particles and tracking them with lasers as they proceed towards the center of the source region of a dual-polarity time-of-flight mass spectrometer. Upon their arrival, they are desorbed and ionized by a single, pulsed high power laser and the resulting ions are measured by the mass spectrometer. Real-time analysis of the mass signature allows the identification of the particles. In the case of many microorganisms, they can be identified to the species level. A SPAMS system can evaluate dozens to hundreds of particles per second, collecting 1000 mass spectra, each of an individual microorganism, in 90 seconds. In this application, we describe the evaluation of SPAMS for the detection of M. tuberculosis from coughed aerosol of TB suspects. M. tuberculosis has been detected in the aerosol phase in clinical settings previously but such detection was performed labor-intensively using the polymerase chain reaction (PCR) to analyze samples collected over a matter of hours [Chen 2005]. We will prove that the diagnosis of TB in real- time is a viable concept by rapidly and accurately detecting M. tuberculosis using this state-of-the-art technology. This is a transformative concept with many potential applications, including rapid triage in hospitals at the emergency room or at TB control clinics to identify patients in need of treatment and to prevent nosocomial transmission, as well as for rapid mass screening in non-clinical environments such as in the field in high incidence settings. Finally, another key advantage of this technology that distinguishes it from other breath analysis systems under development is the ability to train SPAMS for the detection of other respiratory pathogens as well. This makes SPAMS ideal as a diagnostic in TB patients in whom parenchymal damage and bronchiectasis often result in concomitant infection/colonization with other non-TB pathogens. The ability to diagnose other respiratory diseases further broadens SPAMS' potential use in a variety of clinical and non-clinical settings. Livermore Instruments Inc. is a San Francisco Bay Area startup company dedicated to bringing aerosol analysis to nontraditional fields.

描述（由申请人提供）：尽管结核病（TB）造成了人员损失，但这种疾病仍然难以诊断，从而难以控制。世界上有三分之一的人口感染了M。结核每年有900多万人发展为活动性结核病，170万人死于该疾病[WHO 2010]。目前广泛使用的诊断方法在识别活动性TB患者方面表现不佳，大约50%的患者患有抗酸杆菌（AFB）涂片阴性的结核病[Steingart 2006]。未确诊的病例在不知不觉中继续传播疾病，威胁公共卫生。在美国，诊断的不确定性以及缺乏敏感、特异和快速的检测，导致住院费用显著增加，因为疾病控制和预防中心（CDC）指南建议将疑似TB病例隔离在单人床、负压室中，直到连续三次每日痰液样本通过直接涂片处理并被视为AFB阴性[CDC 2005]。由于后勤问题，这些病人平均要隔离五天才能出院。由于这些患者中的绝大多数没有患病，住院患者的结核病控制成本主要由结核病疑似患者的住院隔离天数决定[Scott 1994]，并且通常最终由医院本身承担。与这种感染控制方法相结合的是，AFB阴性痰涂片不能明确排除TB传播的风险[Behr 1999]。当未确诊病例继续传播疾病时，社会成本急剧增加[米勒2010]。目前结核病诊断金标准的局限性激发了人们对快速、准确、成本效益高、非侵入性和用户友好的结核分枝杆菌鉴定方法的新兴趣。结核病嫌疑人的肺结核患者咳嗽气溶胶分析M。结核病有可能满足所有这些要求。 利弗莫尔仪器公司独特的单粒子气溶胶质谱（SPAMS）技术可能是临床现实环境中结核病诊断的答案。SPAMS以前曾被能源部部署用于检测B。炭疽病在这项应用中，我们描述了一个基于SPAMS的气溶胶分析系统作为一个潜在的工具，用于检测M。咳嗽和呼出气中的肺结核。我们现有的系统版本操作简单、坚固、包含很少的移动部件，而且重要的是，不含试剂，因此每次测试的边际成本极低。SPAMS的功能是吸引单个气溶胶粒子，并在它们向双极性飞行时间质谱仪的源区中心移动时用激光跟踪它们。当它们到达时，它们被解吸并被单个脉冲高功率激光电离，并且所产生的离子由质谱仪测量。对质量特征的实时分析允许识别颗粒。对于许多微生物，可以在物种水平上对其进行识别。SPAMS系统每秒可以评估数十到数百个粒子，在90秒内收集1000个质谱，每个质谱都是单个微生物的质谱。 在这个应用中，我们描述了评估的SPAMS检测M。肺结核可疑者咳嗽的气溶胶。M.以前在临床环境中的气溶胶阶段检测到结核病，但这种检测是使用聚合酶链反应（PCR）进行的，需要花费大量的劳动力来分析在几个小时内收集的样本[Chen 2005]。我们将通过快速准确地检测结核分枝杆菌来证明结核病的真实的实时诊断是一个可行的概念。用这种最先进的技术治疗肺结核这是一个具有许多潜在应用的变革性概念，包括医院急诊室或结核病控制诊所的快速分诊， 确定需要治疗的病人，防止医院传播，以及在非临床环境中，如在高发环境中的现场进行快速大规模筛查。最后，该技术与其他正在开发的呼吸分析系统的另一个关键优势是能够训练SPAMS检测其他呼吸道病原体。这使得SPAMS成为肺实质损伤和支气管扩张经常导致其他非结核病原体伴随感染/定植的结核患者的理想诊断方法。诊断其他呼吸系统疾病的能力进一步拓宽了SPAMS在各种临床和非临床环境中的潜在用途。利弗莫尔仪器公司 是一家位于弗朗西斯科湾区的初创公司，致力于将气溶胶分析带入非传统领域。