期刊
ANALYTICAL BIOCHEMISTRY
卷 425, 期 1, 页码 54-61出版社
ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.ab.2012.02.040
关键词
Metal-enhanced fluorescence; Surface-enhanced fluorescence; Plasmon-enhanced fluorescence; Plasmon-controlled fluorescence; Point-of care diagnostics; Anthrax; Protective antigen (PA); Pathogen detection; Bio-warfare agent detection
资金
- Biological Defense Research Directorate, Naval Medical Research Center [00000123002]
- Institute of Fluorescence (IoF) at the University of Maryland, Baltimore County
- Les Baillie (Cardiff University, UK)
Rapid presymptomatic diagnosis of Bacillus anthracis at early stages of infection plays a crucial role in prompt medical intervention to prevent rapid disease progression and accumulation of lethal levels of toxin. To detect low levels of the anthrax protective antigen (PA) exotoxin in biological fluids, we have developed a metal-enhanced fluorescence (MEF)-PA assay using a combination of the MEF effect and microwave-accelerated PA protein surface absorption. The assay is based on a modified version of our rapid catch and signal (RCS) technology previously designed for the ultra-fast and sensitive analysis of genomic DNA sequences. Technologically, the proposed MEF-PA assay uses standard 96-well plastic plates modified with silver island films (SiFs) grown within the wells. It is shown that the fluorescent probe, covalently attached to the secondary antibody, plays a crucial role of indicating complex formation (i.e., shows a strong MEF response to the recognition event). Microwave irradiation rapidly accelerates PA deposition onto the surface (rapid catch), significantly speeding up the MEF-PA assay and resulting in a total assay run time of less than 40 min with an analytical sensitivity of less than 1 pg/ml PA. (C) 2012 Elsevier Inc. All rights reserved.
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