Development of a method for visual detection of Mycobacterium tuberculosis complex

结核分枝杆菌复合群视觉检测方法的开发

• 批准号: 9245274
• 负责人: Yulia Gerasimova
• 金额: $ 20.53万
• 依托单位: UNIVERSITY OF CENTRAL FLORIDA
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-12-15 至 2018-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9245274
• 关键词: Acid Fast Bacillae Staining Method; Address; Adopted; Antibiotic Resistance; Antibiotics; Antitubercular Agents; Bacillus (bacterium); Bacteria; Bacterial Antibiotic Resistance; Bacterial Genes; Bathing; Biological Assay; Caring; Catalytic DNA; Categories; Cells; Cessation of life; Color; Communicable Diseases; Complex; Country; Cytolysis; DNA; Data; Detection; Development; Devices; Diagnosis; Diagnostic; Disease; Disease Outcome; Drug resistance; Ensure; Equipment; Eye; Future; Genes; Goals; Health; Healthcare; Human; Infection; Mediating; Methods; Microscopy; Molecular; Multi-Drug Resistance; Multidrug-Resistant Tuberculosis; Mutation; Mycobacterium bovis; Mycobacterium tuberculosis; Nanotechnology; National Institute of Allergy and Infectious Disease; Nucleic Acids; Outcome; Patient Dropouts; Patients; Pharmaceutical Preparations; Phenotype; Point Mutation; Predisposition; Preparation; Public Health; Pulmonary Tuberculosis; RNA; Recovery; Reporter; Resistance; Resources; Ribosomal RNA; Rifampin; Sampling; Signal Transduction; Solid; Source; Sputum; Technology; Testing; Time; Tuberculosis; Visit; Visual; Water; base; blind; chemotherapy; design; effective therapy; high reward; high risk; improved; isoniazid; novel diagnostics; pathogen; point of care; sensor; tool; tuberculosis drugs; user-friendly

Project Summary Tuberculosis (TB), an infectious disease caused by species of Mycobacterium tuberculosis complex (MTC), remains one of the major public health problems worldwide, with nine million new cases and about 1.5 million deaths each year. In addition, improper use of antibiotics in chemotherapy of drug-susceptible TB patients results in anti-TB drug resistance, which complicates TB care and control. About 95% of lethal cases occur in resource-limited high-burden countries, where there is a lack of diagnostic capacity. Undiagnosed or improperly diagnosed patients develop more severe disease and remain a source of infection for the wider public. At the same time, efficient point-of-care (POC) TB diagnostics in low resource settings can improve health outcomes by facilitating more effective treatment, reducing the patient dropout rate and limiting spread of the infection. The tests of such format are lacking for drug susceptibility testing (DST) of TB. Therefore, new diagnostic tools that are simple enough to be used “in the field” and still be able to correctly diagnose for TB and identify drug-resistance are needed. The goal of this high-risk, high-reward exploratory project is to take advantage of the recent advancements in DNA nanotechnology and molecular sensing to significantly improve diagnostics capabilities for TB. We propose to develop POC-compatible highly selective sensors based on cascades of deoxyribozymes with visual signal readout for detection of active pulmonary TB and testing for mutations conferring multidrug resistance (MDR). Visual deoxyribozyme cascade sensors will be able to produce highly reliable results within the period of usual doctor's visit, and be able to detect less than 104 cells of MTC species without the need for expensive equipment. We will design cascade sensors targeting MTC rRNA and fragments of the genes containing point mutations conferring resistance to the most potent first-line anti-TB drugs rifampicin and isoniazid. We will optimize the assay using isolated bacterial nucleic acids, whole bacterial cells and bacteria-spiked human sputum samples. We will compare the assay with acid fast smear microscopy and culture-based tests, which are currently used for TB detection and DST in the resource-limited settings. The technology proposed here is promising for diagnostics of TB, as well as other pathogens, particularly in resource-constrained high-burden countries, which may revolutionize healthcare worldwide.

项目摘要 结核病（TB），这是由结核分枝杆菌造成的传染病 复杂（MTC）仍然是全球主要的公共卫生问题之一，有900万 病例和每年约150万人死亡。此外，在不当使用抗生素 药物启示性结核病患者的化学疗法会导致抗TB耐药性，这 使结核病护理和控制复杂化。大约95％的致命病例发生在资源有限的高负荷中 国家，缺乏诊断能力。未被诊断或不当诊断 患者患上更严重的疾病，仍然是更广泛的公众感染的根源。 同时，低资源设置中的高效护理（POC）结核病诊断可以改善 通过支持更有效的治疗，降低患者辍学率和 限制感染的传播。这种格式的测试缺乏药物敏感性测试 TB的（DST）。因此，新的诊断工具足够简单，可以“在现场”和 仍然需要正确诊断结核病并识别抗药性。目标的目标 高风险，高回报的探索项目是利用最近的进步 DNA纳米技术和分子传感可显着提高诊断能力 TB。我们建议根据基于POC兼容的高度选择性传感器 具有视觉信号读数的脱氧核酶，用于检测活性肺结核和测试 突变会议会议多药抵抗（MDR）。视觉脱氧核酶级联传感器将 能够在通常的医生访问期间产生高度可靠的结果，并成为 无需昂贵的设备即可检测到少于104个MTC物种的细胞。 我们将设计针对MTC rRNA的级联传感器和包含点基因的片段 突变会议会议对最有效的一线抗TB药物利福平和异烟肼的抗性。 我们将使用孤立的细菌核酸，整个细菌细胞和 细菌尖刺的人类痰液样品。我们将将测定与酸快速涂片进行比较 显微镜和基于培养的测试，目前用于TB检测和DST 资源有限的设置。此处提出的技术对结核病的诊断很有希望 以及其他病原体，特别是在资源受限的高负担国家中，这可能 全世界革新医疗保健。