Nanotechnology-Based Detection of Mycoplasma pneumoniae

基于纳米技术的肺炎支原体检测

• 批准号: 8369123
• 负责人: DUNCAN C KRAUSE
• 金额: $ 22.23万
• 依托单位: UNIVERSITY OF GEORGIA
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-08-15 至 2014-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8369123
• 关键词: Accounting; Advanced Development; Asthma; Biosensing Techniques; Biosensor; Blinded; Bronchitis; Child; Chlamydophila pneumoniae; Chronic; Chronic Obstructive Airway Disease; Chronic lung disease; Clinical; Communities; Complex; Data; Deposition; Detection; Development; Diagnosis; Diagnostic tests; Disease; Electromagnetic Fields; Electromagnetics; Emerging Technologies; Exhibits; False Negative Reactions; Goals; Human; Infection; Infectious Agent; Lead; Lung; Methods; Moraxella (Branhamella) catarrhalis; Morbidity - disease rate; Mycoplasma; Mycoplasma pneumoniae; Nanotechnology; Pattern Recognition; Pharyngeal structure; Physicians; Pneumonia; Publishing; Raman Spectrum Analysis; Respiratory System; Respiratory tract structure; Sampling; Sensitivity and Specificity; Serologic tests; Serological; Serum; Silver; Simulate; Specimen; Streptococcus pyogenes; Surface; Swab; Testing; Virus; Visit; Walking; base; clinical application; clinically relevant; cohort; improved; inhibitor/antagonist; nanofabrication; nanorod; pathogen; rapid diagnosis; secondary infection; transmission process; vapor; young adult

DESCRIPTION (provided by applicant): Mycoplasma pneumoniae is a significant human respiratory tract pathogen, causing bronchitis and atypical or "walking" pneumonia. M. pneumoniae accounts for 20% of all community-acquired pneumonia and is the leading cause of pneumonia in older children and young adults. Serologic testing is the primary method for diagnosis due to the significant challenges posed by direct culture but suffers from severe limitations, including the need for paired sera obtained at separate physician visits, and is thus impractical for rapid diagnosis. PCR can exhibit high sensitivity and yield positive detection sooner but is prone to false-negatives from reaction inhibitors. The inability to provide rapid and definitive diagnosis delays initiation of appropriate treatment, prolongs morbidity, and increases the likelihood of continued transmission, secondary infections, and long-term sequelae, including chronic lung disease associated with COPD and asthma. Lack of a simple, reliable, rapid diagnostic test is thus a critical barrier to improved control of M. pneumoniae disease. Application of nanotechnology to biosensor development is yielding direct, rapid, and sensitive pattern-recognition approaches for detection of infectious agents. We have shown that nanofabrication by glancing angle vapor deposition produces Ag nanorod arrays (NA) exhibiting extremely high electromagnetic field enhancements for surface-enhanced Raman spectroscopy (SERS). Paired with chemometric analysis this platform can rapidly detect and distinguish with outstanding sensitivity and specificity the Raman spectra of viruses and mycoplasmas, and shows great promise in its potential to improve diagnosis of M. pneumoniae infections. Our overall goal is to advance the application of NA-SERS to rapid and sensitive mycoplasma detection in clinical specimens, building upon critical published data that demonstrate its capacity to detect M. pneumoniae at clinically relevant sensitivity in simulated and true clinical throat swab samples. Our application has two specific aims, shifting the project from proof-of-concept toward clinical application: (1) expand analysis of spiked and control samples using throat swabs from a broader cohort, including direct comparison to qPCR; analyze spectral signatures of commensal mycoplasmas and assess their impact on M. pneumoniae detection; and assess NA- SERS discriminatory ability in mixed infections with other bacterial pathogens including Chlamydophila pneumoniae; and (2) test by NA-SERS, true clinical throat swab samples previously established as positive or negative for M. pneumoniae by PCR and culture, in unblinded and blinded analyses; and characterize M. pneumoniae isolates obtained globally to determine whether strain diversity impacts detection by NA-SERS in spiked throat swab samples. PUBLIC HEALTH RELEVANCE: Mycoplasma pneumoniae is a significant human respiratory tract pathogen, accounting for 20- 30% of all community-acquired pneumonia and the leading cause of pneumonia in older children and young adults. The inability to provide rapid and definitive diagnosis delays initiation of appropriate treatment, prolongs morbidity, and increases the likelihood of continued transmission and long-term complications, including COPD and asthma. Lack of a simple and reliable rapid diagnostic test is thus a critical barrier to improved control of M. pneumoniae disease, and our goal is to advance the development of a nanotechnology-based platform for sensitive and expedient detection of M. pneumoniae in clinical specimens.

描述（由申请人提供）：肺炎支原体是一种重要的人类呼吸道病原体，引起支气管炎和非典型或“行走”肺炎。肺炎支原体占所有社区获得性肺炎的 20%，是大龄儿童和年轻人肺炎的主要原因。由于直接培养带来的重大挑战，血清学检测是诊断的主要方法，但也存在严重的局限性，包括需要在单独的医生就诊时获得配对血清，因此对于快速诊断来说是不切实际的。 PCR 可以表现出高灵敏度并更快地产生阳性检测，但很容易因反应抑制剂而出现假阴性。无法提供快速和 明确的诊断会延迟适当治疗的开始，延长发病时间，并增加持续传播、继发感染和长期后遗症的可能性，包括与慢性阻塞性肺病和哮喘相关的慢性肺病。因此，缺乏简单、可靠、快速的诊断测试是改善肺炎支原体疾病控制的关键障碍。纳米技术在生物传感器开发中的应用正在产生直接、快速和灵敏的模式识别方法来检测传染原。我们已经证明，通过掠射角气相沉积进行纳米加工可产生银纳米棒阵列 (NA)，该阵列对表面增强拉曼光谱 (SERS) 具有极高的电磁场增强效果。与化学计量分析相结合，该平台能够以出色的灵敏度和特异性快速检测和区分病毒和支原体的拉曼光谱，并在改善肺炎支原体感染的诊断方面显示出巨大的潜力。我们的总体目标是推进 NA-SERS 在临床样本中快速、灵敏的支原体检测的应用，以已发表的关键数据为基础，这些数据证明了 NA-SERS 在模拟和真实临床咽拭子样本中以临床相关灵敏度检测肺炎支原体的能力。我们的应用有两个具体目标，将项目从概念验证转向临床应用：(1) 使用来自更广泛队列的咽拭子扩大对加标样品和对照样品的分析，包括与 qPCR 的直接比较；分析共生支原体的光谱特征并评估其对肺炎支原体检测的影响；并评估 NA-SERS 对与其他细菌病原体（包括肺炎衣原体）混合感染的区分能力； (2) 通过 NA-SERS 进行测试，即之前在非盲和盲分析中通过 PCR 和培养确定为肺炎支原体阳性或阴性的真实临床咽拭子样本；并对全球获得的肺炎支原体分离株进行表征，以确定菌株多样性是否影响 NA-SERS 在尖刺咽拭子样本中的检测。 公共卫生相关性：肺炎支原体是一种重要的人类呼吸道病原体，占所有社区获得性肺炎的 20-30%，也是年龄较大儿童和年轻人肺炎的主要原因。无法提供快速和明确的诊断会延迟适当治疗的开始，延长发病时间，并增加持续传播和长期并发症（包括慢性阻塞性肺病和哮喘）的可能性。因此，缺乏简单可靠的快速诊断测试是改善肺炎支原体疾病控制的关键障碍，我们的目标是推进基于纳米技术的平台的开发，以灵敏、便捷地检测临床标本中的肺炎支原体。