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），一种由结核分枝杆菌引起的传染病 复杂的（MTC），仍然是全球主要的公共卫生问题之一，有900万新 每年约有150万人死亡。此外，抗生素使用不当， 对药物敏感的结核病患者进行化疗导致抗结核药物耐药性， 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担地区， 缺乏诊断能力的国家。未诊断或诊断不当 病人病情会更严重，仍然是广大公众的感染源。在 与此同时，在低资源环境中进行有效的床旁（POC）结核病诊断可以得到改善 通过促进更有效的治疗，降低患者辍学率， 限制感染的扩散。这种形式的测试缺乏药物敏感性测试 (DST)结核病因此，新的诊断工具，是简单到足以使用“在外地”， 仍然需要能够正确诊断结核病和鉴定耐药性。这个目标 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显著提高了诊断能力， TB.我们建议开发POC兼容的高选择性传感器的基础上级联 用于检测活动性肺结核和测试 多药耐药（MDR）突变。视觉脱氧核酶级联传感器将 能够在通常的医生就诊期间产生高度可靠的结果，并且 能够检测少于104个MTC种类的细胞，而不需要昂贵的设备。 我们将设计针对MTC rRNA和含有点的基因片段的级联传感器， 突变导致对最有效的一线抗结核药物利福平和异烟肼产生耐药性。 我们将使用分离的细菌核酸、全细菌细胞和 加有细菌的人类痰液样本我们将与抗酸涂片进行比较 显微镜和基于培养的测试，目前用于结核病检测和DST在 资源有限的环境。这里提出的技术对于结核病的诊断是有希望的，因为 以及其他病原体，特别是在资源有限的高负担国家， 彻底改变全球医疗保健。