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）：支原体肺炎是一种重要的人类呼吸道病原体，导致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％，是大儿童和年轻人肺炎的主要原因。血清学测试是诊断的主要方法，这是由于直接培养带来的重大挑战，但受到严重限制，包括需要在单独的医师就诊时获得的配对血清，因此对于快速诊断而言是不切实际的。 PCR可以表现出较高的灵敏度并早日产生阳性检测，但容易发生反应抑制剂的假阴性。无法提供快速和 确定的诊断延迟了适当的治疗，延长发病率，增加了持续传播，继发感染和长期后遗症的可能性，包括与COPD和COPD和哮喘相关的慢性肺疾病。因此，缺乏简单，可靠，快速的诊断测试是改善对肺炎支原体疾病的控制的关键障碍。纳米技术在生物传感器开发中的应用正在产生直接，快速和敏感的模式识别方法，以检测传染剂。我们已经表明，通过扫视角度蒸气沉积的纳米化产生纳米棒阵列（NA），表现出极高的电磁场增强，用于表面增强的拉曼光谱（SERS）。与化学计量分析相结合，该平台可以快速检测和区分出出色的敏感性和特异性病毒和支原体的拉曼光谱，并且在改善肺炎支原体感染诊断的潜力方面表现出了巨大的希望。我们的总体目标是在临床标本中促进NA-S在临床标本中的快速和敏感的支原体检测中的应用，这是基于关键已发表的数据，证明了其在模拟和真正的临床喉咙拭子样品中以临床相关的敏感性检测肺炎支原体的能力。我们的应用具有两个具体的目标，将项目从概念验证转变为临床应用：（1）使用更广泛的队列中的喉咙拭子扩展了尖峰和控制样品的分析，包括直接比较qPCR；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；并评估与其他细菌病原体混合感染中的歧视能力，包括肺炎衣原体； （2）通过Na-s的测试，在未盲和盲目的分析中，先前以PCR和培养为肺炎和培养为阳性或阴性的真实临床喉咙样品；并表征全球获得的肺炎开发株分离株，以确定应变多样性是否会影响尖刺的喉咙拭子样品中NA-SER的检测。 公共卫生相关性：支原体肺炎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％，是大儿童和年轻人的肺炎的主要原因。无法提供快速有明确的诊断延迟适当治疗，延长发病率，并增加了持续传播和长期并发症（包括COPD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对肺炎支原体疾病的控制的关键障碍，我们的目标是促进基于纳米技术的基于纳米技术的平台，以敏感且方便地检测临床标本中的肺炎。