Lab-on-a-Film Multiplexed Test for Respiratory Mycobacterial Infections

呼吸道分枝杆菌感染的胶片实验室多重检测

基本信息

批准号：
10256602
负责人：
Christopher Gerard Cooney
金额：
$ 29.96万
依托单位：
AKONNI BIOSYSTEMS, INC.
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-04-21 至 2022-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10256602
关键词：
Address Algorithms Aminoglycosides Antibiotic Resistance Antimicrobial Resistance Antimicrobial susceptibility Bacillus Biological Assay Bronchiectasis Cause of Death Chronic Obstructive Airway Disease Chronic lung disease Clinical Combination Drug Therapy Complex DNA DNA Sequence Alteration Detection Diagnosis Diagnostic Diagnostic radiologic examination Diagnostic tests Disease Drug resistance Drug resistance in tuberculosis Epidemic Etiology Film Genetic Polymorphism Genomics Genus Mycobacterium Glass Incidence Individual Infection Infectious Agent Laboratories Low Prevalence Lung diseases Lung infections Macrolide-resistance Magnetism Methodology Molecular Mycobacterium Infections Mycobacterium tuberculosis Nature Nucleic Acids Organism Pathogenicity Patients Pharmaceutical Preparations Phase Prevalence Process Protocols documentation Rapid diagnostics Resistance Resistance to infection Respiratory Tract Infections Risk Factors Sampling Specimen Sputum Step Tests Testing Time Translating Tuberculosis antimicrobial clinically relevant design detection test microbial multiplex assay mycobacterial non-tuberculosis mycobacteria novel therapeutics prototype rapid test respiratory virtual

项目摘要

ABSTRACT Chronic lung disease due to non-tuberculous mycobacteria (NTM-LD) is a silent and emerging epidemic in the U.S. and many parts of the world.1-3 The incidence and prevalence of NTM-LD is increasing yearly and now surpasses that of tuberculosis (TB) in the U.S.4 Two of the greatest known risk factors for NTM-LD are chronic obstructive pulmonary disease (COPD) and pre-existing bronchiectasis.3 In the U.S., approximately 12 million individuals have COPD, and it is the third leading cause of death in the U.S.5 In addition, the co-occurrence of bronchiectasis in patients with known COPD is estimated to be up to ~70%.6 Like TB, the requirement for prolonged combination drug therapy is a central tenet of NTM treatment. Consequently, it is essential that several drugs be administered concurrently to maximize sterilizing conditions. Unlike TB for which there are now rapid molecular tests for detection of infection and antimicrobial resistance and recently approved new drugs, diseases caused by NTM are a major concern for multiple reasons, foremost being their intrinsic resistance to most existing antimicrobials in the absence of a rapid diagnostic test for antimicrobial susceptibility or even NTM infection.7,8 Diagnosis of NTM-LD is complicated by the fact that clinical manifestations and radiographic findings for TB and NTM-LD are virtually indistinguishable. Thus, it is important when diagnosing NTM-LD to “rule out” TB even in regions of lower prevalence, such as the U.S. because of its infectious and deadly nature. To differentiate TB from NTM-LD, clinicians must rely on a combination of mycobacteriology and molecular tests to identify the etiological agent of the disease, which for NTM includes 13 clinically-relevant species/sub-species that must be distinguished from approximately 180 other NTM species that are rarely pathogenic. In addition to the challenge of accurately speciating mycobacteria, there is also the need to detect antibiotic resistance. Hence, the algorithm for mycobacterial diagnostic testing is complex, requiring varied testing methodologies, which are either slow to respond (culture), insensitive (smears), or insufficiently comprehensive (molecular). Molecular detection of respiratory mycobacterial infections and resistance to antibiotics is challenging. The specimen type (sputum) is viscous and highly heterogeneous; the mycobacteria bacilli are difficult to lyse; the number of species is considerable; and the polymorphisms that confer drug resistance are numerous. To address these challenges, we propose to automate and integrate the following into a one user-step test: chaotic mixing of glass beads using a rotating magnetic disc to homogenize sputum and lyse bacilli, a porous disc in a pipette tip to purify and concentrate nucleic acid, and a Lab-on-a-Film test to speciate and detect polymorphisms that confer drug resistance. For Phase 1, we propose to develop a Lab-on-a-Film test that has the required sensitivity to rule in/out TB, diagnose NTM-LD, and detect drug resistance when Mycobacterium tuberculosis (MTB) and/or clinically important NTM are present.

抽象的非结核分枝杆菌（NTM-LD）引起的慢性肺病是一种无声的、新出现的流行病美国和世界许多地区。1-3 NTM-LD 的发病率和患病率逐年增加，目前在美国超过了结核病 (TB) 4 NTM-LD 的两个已知最大危险因素是慢性阻塞性肺病 (COPD) 和已有的支气管扩张症。3 在美国，大约 1200 万人个人患有慢性阻塞性肺病 (COPD)，它是美国第三大死因。5 此外，同时发生以下疾病已知患有慢性阻塞性肺病 (COPD) 的患者中，支气管扩张的比例估计高达约 70%。6 与结核病一样，需要长期联合药物治疗是经过测试的 NTM 治疗的核心原则，这一点至关重要。与结核病不同的是，几种药物同时施用以最大限度地提高灭菌条件。现在用于检测感染和抗菌素耐药性的快速分子检测以及最近批准的新方法由于多种原因，由 NTM 药物引起的疾病成为一个主要问题，最重要的是其内在的原因在缺乏抗菌药物快速诊断测试的情况下，对大多数现有抗菌药物产生耐药性易感性甚至 NTM 感染。7,8 NTM-LD 的诊断因临床表现而变得复杂 TB 和 NTM-LD 的表现和影像学表现实际上无法区分。即使在美国等流行率较低的地区，诊断 NTM-LD 以“排除”结核病时也很重要。由于其传染性和致命性，要将 TB 与 NTM-LD 区分开来，必须依靠结合分枝杆菌学和分子测试来确定疾病的病原体，这对于 NTM 包括 13 个临床相关物种/亚种，必须与大约 180 个物种/亚种区分开来除了准确鉴定物种的挑战外，其他很少致病的 NTM 物种也面临着挑战。分枝杆菌，还需要检测抗生素耐药性，因此，分枝杆菌的算法。诊断测试复杂、多样，需要测试方法，而这些方法要么响应缓慢（培养）、不敏感（涂片）或不够全面（分子）。呼吸道分枝杆菌感染和抗生素耐药性的分子检测具有挑战性。标本类型（痰）粘稠且高度异质；分枝杆菌难以裂解；物种数量相当多；并且赋予耐药性的多态性也很多。为了应对这些挑战，我们建议将以下内容自动化并集成到一个用户步骤测试中：使用旋转磁盘对玻璃珠进行混乱混合，以均化痰液和裂解杆菌（一种多孔的细菌）移液器吸头中的圆盘用于纯化和浓缩核酸，以及用于物种形成和检测的薄膜实验室测试对于第一阶段，我们建议开发一种薄膜实验室测试。排除/排除结核病、诊断 NTM-LD 和检测分枝杆菌耐药性所需的灵敏度存在结核病 (MTB) 和/或临床上重要的 NTM。