Fluorescent Nanoparticle-Based Indirect Immunofluorescence Microscopy for Detection 
      of <em>Mycobacterium tuberculosis</em>

doi:10.1155/2007/89364

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Journal of Biomedicine and Biotechnology
Volume 2007 (2007), Article ID 89364, 9 pages
doi:10.1155/2007/89364

Research Article

Fluorescent Nanoparticle-Based Indirect Immunofluorescence Microscopy for Detection of Mycobacterium tuberculosis

Dilan Qin,^1,2 Xiaoxiao He,^1,2 Kemin Wang,^1,2 Xiaojun Julia Zhao,³ Weihong Tan,^1,2 and Jiyun Chen^1,2

¹State Key Laboratory of Chemo/Biosensing and Chemometrics, Biomedical Engineering Center, College of Chemistry and Chemical Engineering, Hunan University, Changsha, Hunan 410082, China
²Key Laboratory for Bio-Nanotechnology and Molecule Engineering of Hunan Province, Hunan University, Changsha 410082, China
³Department of Chemistry, University of North Dakota, Grand Forks 58202, ND, USA

Received 24 March 2007; Revised 10 July 2007; Accepted 10 October 2007

Academic Editor: Marek Osinski

Copyright © 2007 Dilan Qin et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

A method of fluorescent nanoparticle-based indirect immunofluorescence microscopy (FNP-IIFM) was developed for the rapid detection of Mycobacterium tuberculosis. An anti-Mycobacterium tuberculosis antibody was used as primary antibody to recognize Mycobacterium tuberculosis, and then an antibody binding protein (Protein A) labeled with Tris(2,2-bipyridyl)dichlororuthenium(II) hexahydrate (RuBpy)-doped silica nanoparticles was used to generate fluorescent signal for microscopic examination. Prior to the detection, Protein A was immobilized on RuBpy-doped silica nanoparticles with a coverage of ∼5.1×102 molecules/nanoparticle. With this method, Mycobacterium tuberculosis in bacterial mixture as well as in spiked sputum was detected. The use of the fluorescent nanoparticles reveals amplified signal intensity and higher photostability than the direct use of conventional fluorescent dye as label. Our preliminary studies have demonstrated the potential application of the FNP-IIFM method for rapid detection of Mycobacterium tuberculosis in clinical samples.