Nanotechnology-Based Detection of Mycoplasma pneumoniae

基于纳米技术的肺炎支原体检测

• 批准号: 8522149
• 负责人: DUNCAN C KRAUSE
• 金额: $ 19.07万
• 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 2015-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8522149
• 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.

描述(申请人提供)：肺炎支原体是一种重要的人类呼吸道病原体，可引起支气管炎和非典型肺炎或“行走”肺炎。肺炎支原体占所有社区获得性肺炎的20%，是年龄较大的儿童和年轻人肺炎的主要原因。血清学检测是诊断的主要方法，因为直接培养带来了巨大的挑战，但受到严重限制，包括需要在单独的医生会诊时获得配对血清，因此不适用于快速诊断。聚合酶链式反应具有较高的灵敏度和较快的阳性检测速度，但容易出现反应抑制物的假阴性。无法提供快速和 明确的诊断推迟了适当治疗的开始，延长了发病率，并增加了继续传播、继发感染和长期后遗症的可能性，包括与COPD和哮喘相关的慢性肺部疾病。因此，缺乏一种简单、可靠、快速的诊断试验是改善肺炎支原体疾病控制的关键障碍。将纳米技术应用到生物传感器的开发中，正在产生直接、快速和灵敏的模式识别方法来检测感染性病原体。我们已经证明，通过掠角气相沉积进行纳米加工可以得到具有极高电磁场增强的银纳米棒阵列(NA)，用于表面增强拉曼光谱(SERS)。与化学计量学分析相结合，该平台可以快速检测和区分病毒和支原体的拉曼光谱，具有极高的灵敏度和特异性，在提高肺炎支原体感染诊断方面显示出巨大的潜力。我们的总体目标是推动NA-SERS在临床标本中快速和灵敏地检测支原体方面的应用，基于已发表的关键数据，这些数据表明，NA-SERS能够在模拟和真实的临床咽拭子样本中检测临床相关的肺炎支原体。我们的应用有两个具体目标，将项目从概念验证转移到临床应用：(1)扩大使用更广泛队列的咽拭子对添加和对照样本的分析，包括直接与qPCR进行比较；分析共生支原体的光谱特征并评估它们对肺炎支原体检测的影响；以及评估NA-SERS在与其他细菌病原体(包括肺炎衣原体)混合感染中的区分能力；以及(2)NA-SERS检测，在非盲法和盲法分析中，先前经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特征分析，以确定菌株多样性是否影响在添加了添加剂的咽拭子样本中使用NA-SERS进行检测。

项目成果

Highly Sensitive Detection and Differentiation of Endotoxins Derived from Bacterial Pathogens by Surface-Enhanced Raman Scattering.

• DOI: 10.3390/bios11070234
• 发表时间: 2021-07-11
• 期刊: Biosensors
• 作者: Wu X;Zhao Y;Zughaier SM
• 通讯作者: Zughaier SM

Layer-by-layer polyelectrolyte encapsulation of Mycoplasma pneumoniae for enhanced Raman detection.

肺炎支原体的层层聚电解质封装用于增强拉曼检测。

• DOI: 10.1039/c4an00596a
• 发表时间: 2014
• 期刊: The Analyst
• 作者: Rivera-Betancourt,OmarE;Sheppard,EdwardS;Krause,DuncanC;Dluhy,RichardA
• 通讯作者: Dluhy,RichardA

Specificity and Strain-Typing Capabilities of Nanorod Array-Surface Enhanced Raman Spectroscopy for Mycoplasma pneumoniae Detection.

纳米棒阵列表面增强拉曼光谱检测肺炎支原体的特异性和菌株分型能力。

• DOI: 10.1371/journal.pone.0131831
• 发表时间: 2015
• 期刊: PloS one
• 影响因子: 3.7
• 作者: Henderson,KelleyC;Benitez,AlvaroJ;Ratliff,AmyE;Crabb,DonnaM;Sheppard,EdwardS;Winchell,JonasM;Dluhy,RichardA;Waites,KenB;Atkinson,TPrescott;Krause,DuncanC
• 通讯作者: Krause,DuncanC

Culture-free diagnostics of Pseudomonas aeruginosa infection by silver nanorod array based SERS from clinical sputum samples.

• DOI: 10.1016/j.nano.2014.04.010
• 发表时间: 2014-11
• 期刊: NANOMEDICINE-NANOTECHNOLOGY BIOLOGY AND MEDICINE
• 影响因子: 5.4
• 作者: Wu, Xiaomeng;Chen, Jing;Li, Xibo;Zhao, Yiping;Zughaier, Susu M.
• 通讯作者: Zughaier, Susu M.

The multivariate detection limit for Mycoplasma pneumoniae as determined by nanorod array-surface enhanced Raman spectroscopy and comparison with limit of detection by qPCR.

• DOI: 10.1039/c4an01141d
• 发表时间: 2014-12-21
• 期刊: The Analyst
• 作者: Henderson KC;Sheppard ES;Rivera-Betancourt OE;Choi JY;Dluhy RA;Thurman KA;Winchell JM;Krause DC
• 通讯作者: Krause DC